CROSS REFERENCE TO RELATED APPLICATIONS This application is a continuation of U.S. application Ser. No. 16/682,845, filed on Nov. 13, 2019, which is a continuation of the International Application No. PCT/US2019/023046, filed Mar. 19, 2019, which claims the benefit of priority under 35 U.S.C. § 119(e) of U.S. Provisional Patent Application No. 62/645,081, filed Mar. 19, 2018, the disclosure of each of which is incorporated herein by reference in their entireties for all purposes.
INCORPORATION BY REFERENCE OF SEQUENCE LISTING The contents of the text file named “REBI_001_01US_SeqList_ST25.txt”, which was created on Nov. 12, 2019 and is 1,190 megabytes in size, are hereby incorporated by reference in its entirety.
TECHNICAL FIELD The present disclosure is generally related to the biosynthesis of organic compounds, such as cannabinoids, using recombinant enzymes.
BACKGROUND Cannabinoids include a group of more than 100 chemical compounds mainly found in the plant Cannabis sativa L. Due to the unique interaction of cannabinoids with the human endocannabinoid system, many of these compounds are potential therapeutic agents for the treatment of several medical conditions. For instance, the psychoactive compound Δ9-tetrahydrocannabinol (Δ9-THC) has been used in the treatment of pain and other medical conditions. Several synthetic Cannabis-based preparations have been used in the USA, Canada and other countries as an authorized treatment for nausea and vomiting in cancer chemotherapy, appetite loss in acquired immune deficiency syndrome and symptomatic relief of neuropathic pain in multiple sclerosis.
Cannabinoids are terpenophenolic compounds, produced from fatty acids and isoprenoid precursors as part of the secondary metabolism of Cannabis. The main cannabinoids produced by Cannabis are Δ9-tetrahydrocannabidiol (THC), cannabidiol (CBD) and cannabinol (CBN), followed by cannabigerol (CBG), cannabichromene (CBC) and other minor constituents. Currently, Δ9-THC and CBD are either extracted from the plant or chemically synthesized. However, agricultural production of cannabinoids faces challenges such as plant susceptibility to climate and diseases, low content of less-abundant cannabinoids, and need for extraction of cannabinoids by chemical processing. Furthermore, chemical synthesis of cannabinoids has failed to be a cost-effective alternative mainly because of complex synthesis leading to high production cost and low yields.
Therefore, there is a pressing need for biotechnology-based synthetic biology approaches which can enable the synthesis of high-quality cannabinoids in a cost-effective and environmentally friendly manner.
SUMMARY In one aspect, the disclosure provides a recombinant polypeptide comprising an amino acid sequence with at least 95% identity to SEQ ID NO: 1, wherein said amino acid sequence comprises at least one amino acid substitution to SEQ ID NO: 1, wherein said at least one amino acid substitution does not comprise an alanine substitution at a position selected from the group consisting of amino acid positions 47, 64, 110, 121, 123, 126, 161, 175, 177, 214, 216, 288, 294, and 295.
In one aspect, the disclosure provides a recombinant polypeptide comprising an amino acid sequence with at least 95% identity to SEQ ID NO: 1, wherein said amino acid sequence comprises at least one amino acid substitution to SEQ ID NO: 1, wherein at least one of said at least one amino acid substitution to SEQ ID NO: 1 is located on a position chosen from the group consisting of amino acid positions 1-46, 48-63, 65-109, 111-120, 122, 124, 125, 127-160, 162-174, 176, 178-213, 215, 217-287, 289-293, and 296-307.
In some cases, the amino acid sequence has at least 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 1. In some cases, the at least one amino acid substitution comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 amino acid substitutions to SEQ ID NO: 1. In some cases, said at least one of said at least one amino acid substitution is located on a position chosen from the group consisting of amino acid positions 17, 25, 38, 49, 53, 106, 108, 112, 118, 119, 121, 123, 161, 162, 166, 173, 174, 177, 205, 209, 213, 214, 216, 219, 227, 228, 230, 232, 271, 274, 283, 286, 288, 294, 295, and 298. In some cases, said at least one of said at least one amino acid substitution is located on a position chosen from the group consisting of amino acid positions 17, 25, 38, 49, 53, 106, 108, 112, 118, 119, 162, 166, 173, 174, 205, 209, 213, 219, 227, 228, 230, 232, 271, 274, 283, 286, 288, and 298.
In some cases, said at least one of said at least one amino acid substitution is chosen from the group consisting of A17T, C25V, Q38G, V49A, V49L, V49S, A53C, A53D, A53E, A53F, A53G, A53H, A53I, A53K, A53L, A53M, A53N, A53P, A53Q, A53R, A53S, A53T, A53V, A53W, A53Y, M106E, A108G, E112D, E112G, K118N, K118Q, K119A, K119D, Y121W, F123A, F123H, F123W, Q161A, Q161C, Q161D, Q161E, Q161F, Q161G, Q161H, Q161I, Q161K, Q161L, Q161M, Q161N, Q161P, Q161R, Q161S, Q161T, Q161V, Q161W, Q161Y, M162A, M162F, D166E, N173D, L174V, S177E, S177W, S177Y, G205L, G205M, C209G, F213M, S214A, S214C, S214D, S214E, S214F, S214G, S214H, S214I, S214K, S214L, S214M, S214N, S214P, S214Q, S214R, S214T, S214V, S214W, S214Y, Y216A, L219F, D227E, R228E, R228Q, C230N, C230S, A232S, V271E, L274V, Y283L, G286E, Y288A, Y288C, Y288D, Y288E, Y288F, Y288G, Y288H, Y288I, Y288K, Y288L, Y288M, Y288N, Y288P, Y288Q, Y288R, Y288S, Y288T, Y288V, Y288W, V294A, V294F, V294N, Q295A, Q295C, Q295D, Q295E, Q295F, Q295G, Q295H, Q295I, Q295K, Q295L, Q295M, Q295N, Q295P, Q295R, Q295S, Q295T, Q295V, Q295W, Q295Y, L298A, L298Q, and L298W.
In some cases, said recombinant polypeptide converts olivetolic acid (OA) and geranyl diphosphate (GPP) to cannabigerolic acid (CBGA) and 5-geranyl olivetolate (5-GOA), and wherein said recombinant polypeptide produces a ratio of an amount of CBGA to a combined amount of CBGA and 5-GOA that is higher than wild-type ORF2 enzyme under the same condition. In some cases, said ratio is 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 150%, or 200% higher than wild-type ORF2 enzyme under the same condition. In some cases, said at least one amino acid substitution to SEQ ID NO: 1 is selected from the group consisting of Q295F, Q295L, Q295V, Q295I, Q295M, Q295E, Q295C, Q295W, Q295G, Q295T, Q295P, Q295S, Q295D, Q295N, Q295K, and Q295Y. In some cases, said amino acid sequence is selected from the group consisting of SEQ ID NOs: 139-143; 145-150; and 153-157. In some cases, said at least one amino acid substitution to SEQ ID NO: 1 is selected from the group consisting of Q161H, Q161Y, Q161R, Q161D, Q161F, Q161K, Q161M, Q161E, Q161L, Q161S, Q161T, Q161N, and Q161I. In some cases, said amino acid sequence is selected from the group consisting of SEQ ID NOs: 120-122; 124-129; 132-134; and 137. In some cases, said at least one amino acid substitution to SEQ ID NO: 1 is selected from the group consisting of S214H, S214R, S214Q, S214E, S214K, S214M, S214I, S214L, S214N, S214D, S214Y, S214F, and S214T. In some cases, said amino acid sequence is selected from the group consisting of SEQ ID NOs: 160-162; 164-169; 171, 172, 174, and 177. In some cases, said at least one amino acid substitution to SEQ ID NO: 1 is selected from the group consisting of Y288F, Y288P, Y288A, Y288L, Y288C, Y288I, Y288M, Y288G, Y288T, Y288S, Y288W, Y288D, Y288E, Y288N, Y288K, Y288R, and Y288H. In some cases, said amino acid sequence is selected from the group consisting of SEQ ID NOs: 98-110; 112-114; and 116.
In some cases, said at least one amino acid substitution to SEQ ID NO: 1 comprises two or more amino acid substitutions to SEQ ID NO: 1 selected from the group consisting of
(a) A17T, C25V, Q38G, V49A, V49L, V49S, A53C, A53D, A53E, A53F, A53G, A53H, A53I, A53K, A53L, A53M, A53N, A53P, A53Q, A53R, A53S, A53T, A53V, A53W, A53Y, M106E, A108G, E112D, E112G, K118N, K118Q, K119A, K119D, Y121W, F123A, F123H, F123W, Q161A, Q161C, Q161D, Q161E, Q161F, Q161G, Q161H, Q161I, Q161K, Q161L, Q161M, Q161N, Q161P, Q161R, Q161S, Q161T, Q161V, Q161W, Q161Y, M162A, M162F, D166E, N173D, L174V, S177E, S177W, S177Y, G205L, G205M, C209G, F213M, S214A, S214C, S214D, S214E, S214F, S214G, S214H, S214I, S214K, S214L, S214M, S214N, S214P, S214Q, S214R, S214T, S214V, S214W, S214Y, Y216A, L219F, D227E, R228E, R228Q, C230N, C230S, A232S, V271E, L274V, Y283L, G286E, Y288A, Y288C, Y288D, Y288E, Y288F, Y288G, Y288H, Y288I, Y288K, Y288L, Y288M, Y288N, Y288P, Y288Q, Y288R, Y288S, Y288T, Y288V, Y288W, V294A, V294F, V294N, Q295A, Q295C, Q295D, Q295E, Q295F, Q295G, Q295H, Q295I, Q295K, Q295L, Q295M, Q295N, Q295P, Q295R, Q295S, Q295T, Q295V, Q295W, Q295Y, L298A, L298Q, and L298W; or
(b) A53T and S214R; S177W and Q295A; S214R and Q295F; Q161S and 5214R; S177W and 5214R; Q161S and Q295L; Q161S and Q295F; V49A and 5214R; A53T and Q295F; Q161S and S177W; Q161S, V294A and Q295W; A53T, Q161S and Q295W; A53T and S177W; A53T, Q161S, V294A and Q295W; A53T, V294A and Q295A; V49A and Q295L; A53T, Q161S, V294N and Q295W; A53T and Q295A; Q161S, V294A and Q295A; A53T and Q295W; A53T, V294A and Q295W; A53T, Q161S and Q295A; A53T, Q161S, V294A and Q295A; and A53T, Q161S, V294N and Q295A. In some cases, said amino acid sequence is selected from the group consisting of SEQ ID NOs: 275-294; and 296-299.
In some cases, said recombinant polypeptide converts olivetolic acid (OA) and geranyl diphosphate (GPP) to cannabigerolic acid (CBGA) and 5-geranyl olivetolate (5-GOA), and wherein said recombinant polypeptide produces a ratio of an amount of 5-GOA to a combined amount of CBGA and 5-GOA that is higher than wild-type ORF2 enzyme under the same condition. In some cases, said ratio is 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 150%, or 200% higher than wild-type ORF2 enzyme under the same condition. In some cases, said at least one amino acid substitution to SEQ ID NO: 1 is selected from the group consisting of Q295H and Q295R. In some cases, said amino acid sequence is selected from the group consisting of SEQ ID NOs: 144 and 152. In some cases, said at least one amino acid substitution to SEQ ID NO: 1 is selected from the group consisting of Q161W, Q161G, Q161V, Q161C, and Q161P. In some cases, said amino acid sequence is selected from the group consisting of SEQ ID NOs: 118, 119, 123, 130, 131, 135, and 136. In some cases, said at least one amino acid substitution to SEQ ID NO: 1 is selected from the group consisting of S214V, S214C, S214W, S214P, S214A, and S214G. In some cases, said amino acid sequence is selected from the group consisting of SEQ ID NOs: 159, 163, 170, 175 and 176. In some cases, said at least one amino acid substitution to SEQ ID NO: 1 is selected from the group consisting of A53T and Q161S; A53T, Q161S and V294A; A53T, Q161S and V294N; A53T and V294A; and V49A and Q161S. In some cases, said amino acid sequence is selected from the group consisting of SEQ ID NOs: 272-274, 295, and 300.
In some cases, said recombinant polypeptide converts olivetolic acid (OA) and geranyl diphosphate (GPP) to cannabigerolic acid (CBGA) and 5-geranyl olivetolate (5-GOA), and wherein said recombinant polypeptide produces a combined amount of CBGA and 5-GOA that is higher than wild-type ORF2 enzyme under the same condition. In some cases, said combined amount is 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 150%, or 200% higher than wild-type ORF2 enzyme under the same condition. In some cases, said at least one amino acid substitution to SEQ ID NO: 1 is selected from the group consisting of Q295H, Q295F, Q295W, Q295M, Q295D, Q295L, and Q295Y. In some cases, said amino acid sequence is selected from the group consisting of SEQ ID NOs: 140, 142, 144, 147, 148, 156 and 157. In some cases, said at least one amino acid substitution to SEQ ID NO: 1 is selected from the group consisting of Q161H, Q161Y, Q161L, Q161R, Q161C, Q161T, Q161S, Q161F, Q161I, and Q161P. In some cases, said amino acid sequence is selected from the group consisting of SEQ ID NOs: 119, 122, 124, 125, 127, 130, 132-134, and 137. In some cases, said at least one amino acid substitution to SEQ ID NO: 1 is selected from the group consisting of S214G and S214H. In some cases, said amino acid sequence is selected from the group consisting of SEQ ID NOs: 163 and 164. In some cases, said at least one amino acid substitution to SEQ ID NO: 1 is selected from the group consisting of Y288L, Y288P, Y288I, Y288A, Y288F, Y288C, Y288M, and Y288T. In some cases, said amino acid sequence is selected from the group consisting of SEQ ID NOs: 98, 99, 102, 105, 107, 108, 110, and 114.
In some cases, said at least one amino acid substitution to SEQ ID NO: 1 comprises two or more amino acid substitutions selected from the group consisting of:
(a) A17T, C25V, Q38G, V49A, V49L, V49S, A53C, A53D, A53E, A53F, A53G, A53H, A53I, A53K, A53L, A53M, A53N, A53P, A53Q, A53R, A53S, A53T, A53V, A53W, A53Y, M106E, A108G, E112D, E112G, K118N, K118Q, K119A, K119D, Y121W, F123A, F123H, F123W, Q161A, Q161C, Q161D, Q161E, Q161F, Q161G, Q161H, Q161I, Q161K, Q161L, Q161M, Q161N, Q161P, Q161R, Q161S, Q161T, Q161V, Q161W, Q161Y, M162A, M162F, D166E, N173D, L174V, S177E, S177W, S177Y, G205L, G205M, C209G, F213M, S214A, S214C, S214D, S214E, S214F, S214G, S214H, S214I, S214K, S214L, S214M, S214N, S214P, S214Q, S214R, S214T, S214V, S214W, S214Y, Y216A, L219F, D227E, R228E, R228Q, C230N, C230S, A232S, V271E, L274V, Y283L, G286E, Y288A, Y288C, Y288D, Y288E, Y288F, Y288G, Y288H, Y288I, Y288K, Y288L, Y288M, Y288N, Y288P, Y288Q, Y288R, Y288S, Y288T, Y288V, Y288W, V294A, V294F, V294N, Q295A, Q295C, Q295D, Q295E, Q295F, Q295G, Q295H, Q295I, Q295K, Q295L, Q295M, Q295N, Q295P, Q295R, Q295S, Q295T, Q295V, Q295W, Q295Y, L298A, L298Q, and L298W; or
(b) A53T and V294A; A53T, Q161S and V294A; A53T, Q161S and V294N; Q161S, V294A and Q295A; A53T, Q161S and Q295A; A53T, Q161S, V294A and Q295A; A53T, Q161S, V294N and Q295A; A53T and Q295W; Q161S, V294A and Q295W; A53T, Q161S and Q295W; A53T, V294A and Q295W; A53T, Q161S, V294A and Q295W; A53T, Q161S, V294N and Q295W; S177W and Q295A; A53T and S177W; A53T and Q295F; A53T and 5214R; A53T and Q161S; Q161S and Q295F; Q161S and Q295L; and S214R and Q295F. In some cases, said amino acid sequence is selected from the group consisting of SEQ ID NOs: 272-274, 276, 277, 279-287; 290, 293-297, and 299.
In some cases, said recombinant polypeptide converts olivetolic acid (OA) and geranyl diphosphate (GPP) to cannabigerolic acid (CBGA) and 5-geranyl olivetolate (5-GOA), and wherein said recombinant polypeptide has a CBGA production potential that is higher than wild-type ORF2 enzyme under the same condition. In some cases, said CBGA production potential is 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 150%, or 200% higher than wild-type ORF2 enzyme under the same condition. In some cases, said at least one amino acid substitution to SEQ ID NO: 1 is selected from the group consisting of Q295F, Q295M, Q295W, Q295L, Q295I, Q295C, Q295G, Q295V, Q295D, and Q295H. In some cases, said amino acid sequence is selected from the group consisting of SEQ ID NOs: 139, 140, 142-145, 147, 148, 155 and 156. In some cases, said at least one amino acid substitution to SEQ ID NO: 1 is selected from the group consisting of Q161H, Q161Y, Q161R, Q161F, Q161L, Q161S, Q161T, Q161I, Q161M, and Q161K. In some cases, said amino acid sequence is selected from the group consisting of SEQ ID NOs: 122, 124-128, 132-134, and 137. In some cases, said at least one amino acid substitution to SEQ ID NO: 1 is selected from the group consisting of S214H, S214Q, and S214R. In some cases, said amino acid sequence is selected from the group consisting of SEQ ID NOs: 164, 171 and 172. In some cases, said at least one amino acid substitution to SEQ ID NO: 1 is selected from the group consisting of Y288L, Y288P, Y288I, Y288A, Y288F, Y288C, Y288M, and Y288T. In some cases, said amino acid sequence is selected from the group consisting of SEQ ID NOs: 98, 99, 102, 105, 107, 108, 110 and 114. In some cases, said at least one amino acid substitution to SEQ ID NO: 1 comprises two or more amino acid substitutions selected from the group consisting of:
(a) A17T, C25V, Q38G, V49A, V49L, V49S, A53C, A53D, A53E, A53F, A53G, A53H, A53I, A53K, A53L, A53M, A53N, A53P, A53Q, A53R, A53S, A53T, A53V, A53W, A53Y, M106E, A108G, E112D, E112G, K118N, K118Q, K119A, K119D, Y121W, F123A, F123H, F123W, Q161A, Q161C, Q161D, Q161E, Q161F, Q161G, Q161H, Q161I, Q161K, Q161L, Q161M, Q161N, Q161P, Q161R, Q161S, Q161T, Q161V, Q161W, Q161Y, M162A, M162F, D166E, N173D, L174V, S177E, S177W, S177Y, G205L, G205M, C209G, F213M, S214A, S214C, S214D, S214E, S214F, S214G, S214H, 52141, S214K, S214L, S214M, S214N, S214P, S214Q, S214R, S214T, S214V, S214W, S214Y, Y216A, L219F, D227E, R228E, R228Q, C230N, C230S, A232S, V271E, L274V, Y283L, G286E, Y288A, Y288C, Y288D, Y288E, Y288F, Y288G, Y288H, Y288I, Y288K, Y288L, Y288M, Y288N, Y288P, Y288Q, Y288R, Y288S, Y288T, Y288V, Y288W, V294A, V294F, V294N, Q295A, Q295C, Q295D, Q295E, Q295F, Q295G, Q295H, Q295I, Q295K, Q295L, Q295M, Q295N, Q295P, Q295R, Q295S, Q295T, Q295V, Q295W, Q295Y, L298A, L298Q, and L298W; or
(b) A53T_Q161S_V294A; A53T_Q161S_V294N; A53T_Q295A; Q161S_V294A_Q295A; A53T_Q161S_Q295A; A53T_V294A_Q295A; A53T_Q161S_V294A_Q295A; A53T_Q161S_V294N_Q295A; A53T_Q295W; Q161S_V294A_Q295W; A53T_Q161S_Q295W; A53T_V294A_Q295W; A53T_Q161S_V294A_Q295W; A53T_Q161S_V294N_Q295W; S177W_Q295A; S177W_S214R; Q161S_S177W; A53T_S177W; V49A_S214R; A53T_Q295F; A53T_S214R; A53T_Q161S; Q161S_Q295F; Q161S_Q295L; Q161S_S214R; and S214R_Q295F. In some cases, the recombinant polypeptide of any one of claims Error! Reference source not found.-Error! Reference source not found., wherein said amino acid sequence is selected from the group consisting of SEQ ID NOs: 273-290, and 292-299.
In some cases, said recombinant polypeptide converts olivetolic acid (OA) and geranyl diphosphate (GPP) to cannabigerolic acid (CBGA) and 5-geranyl olivetolate (5-GOA), and wherein said recombinant polypeptide has a 5-GOA production potential that is higher than wild-type ORF2 enzyme under the same condition. In some cases, said 5-GOA production potential is 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 150%, or 200% higher than wild-type ORF2 enzyme under the same condition. In some cases, said at least one amino acid substitution to SEQ ID NO: 1 is selected from the group consisting of Q295H and Q295Y. In some cases, said amino acid sequence is selected from the group consisting of SEQ ID NOs: 144 and 157. In some cases, said at least one amino acid substitution to SEQ ID NO: 1 is selected from the group consisting of Q161C, Q161L, Q161T, Q161P, Q161I, and Q161S. In some cases, said amino acid sequence is selected from the group consisting of SEQ ID NOs: 119, 125, 127, 130, 133 and 134. In some cases, said at least one amino acid substitution to SEQ ID NO: 1 is S214G. In some cases, said amino acid sequence is SEQ ID NO: 163. In some cases, said at least one amino acid substitution to SEQ ID NO: 1 is selected from the group consisting of A53T_V294A; V49A_Q295L; and V49A_Q161S. In some cases, said amino acid sequence is selected from the group consisting of SEQ ID NOs: 272, 291, and 300.
In some cases, said recombinant polypeptide converts DVA and geranyl diphosphate (GPP) to CBGVA and 5-GDVA, and wherein said recombinant polypeptide produces a ratio of an amount of CBGVA to a combined amount of CBGVA and 5-GDVA that is higher than wild-type ORF2 enzyme under the same condition. In some cases, said ratio is 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 150%, or 200% higher than wild-type ORF2 enzyme under the same condition. In some cases, said at least one amino acid substitution to SEQ ID NO: 1 is selected from the group consisting of Q295F, Q295I, Q295W, Q295V, Q295M, Q295L, Q295E, Q295H, Q295P, Q295C, Q295T and Q295Y. In some cases, said amino acid sequence is selected from the group consisting of SEQ ID NOs: 139, 141, 142, 144, 145, 147, 148, 150, 154, 155, 156, and 157. In some cases, said at least one amino acid substitution to SEQ ID NO: 1 is selected from the group consisting of Q161H, Q161Y, Q161R, Q161K, Q161E, Q161S, Q161D, Q161F, Q161T, Q161G, and Q161P. In some cases, said amino acid sequence is selected from the group consisting of SEQ ID NOs: 120, 121, 122, 123, 124, 126, 130, 132, 133, 134, and 137. In some cases, said at least one amino acid substitution to SEQ ID NO: 1 is selected from the group consisting of S214R, S214H, S214K, S214E, S214Q, S214L, S214M, S214Y, 52141, S214F, S214D, S214N, S214G, S214T, and S214V. In some cases, said amino acid sequence is selected from the group consisting of SEQ ID NOs: 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 171, 172, 174, 175, and 177. In some cases, said at least one amino acid substitution to SEQ ID NO: 1 is selected from the group consisting of Y288F, Y288I, Y288L, Y288M, Y288P, Y288T, Y288W, Y288C, and Y288G. In some cases, said amino acid sequence is selected from the group consisting of SEQ ID NOs: 99, 102, 103, 105, 107, 108, 110, 114, and 116.
In some cases, said at least one amino acid substitution to SEQ ID NO: 1 comprises two or more amino acid substitution to SEQ ID NO: 1 selected from the group consisting of:
(a) A17T, C25V, Q38G, V49A, V49L, V49S, A53C, A53D, A53E, A53F, A53G, A53H, A53I, A53K, A53L, A53M, A53N, A53P, A53Q, A53R, A53S, A53T, A53V, A53W, A53Y, M106E, A108G, E112D, E112G, K118N, K118Q, K119A, K119D, Y121W, F123A, F123H, F123W, Q161A, Q161C, Q161D, Q161E, Q161F, Q161G, Q161H, Q161I, Q161K, Q161L, Q161M, Q161N, Q161P, Q161R, Q161S, Q161T, Q161V, Q161W, Q161Y, M162A, M162F, D166E, N173D, L174V, S177E, S177W, S177Y, G205L, G205M, C209G, F213M, S214A, S214C, S214D, S214E, S214F, S214G, S214H, 52141, S214K, S214L, S214M, S214N, S214P, S214Q, S214R, S214T, S214V, S214W, S214Y, Y216A, L219F, D227E, R228E, R228Q, C230N, C230S, A232S, V271E, L274V, Y283L, G286E, Y288A, Y288C, Y288D, Y288E, Y288F, Y288G, Y288H, Y288I, Y288K, Y288L, Y288M, Y288N, Y288P, Y288Q, Y288R, Y288S, Y288T, Y288V, Y288W, V294A, V294F, V294N, Q295A, Q295C, Q295D, Q295E, Q295F, Q295G, Q295H, Q295I, Q295K, Q295L, Q295M, Q295N, Q295P, Q295R, Q295S, Q295T, Q295V, Q295W, Q295Y, L298A, L298Q, and L298W; or
(b) A53T_S214R, Q161S_Q295L, S177W_Q295A, Q161S_Q295F, A53T_Q295F, S214R_Q295F, Q161S_S177W, Q16S_S214R, A53T_Q161S, A53T_V294A, A53T_S177W, A53T_Q161S_V294A_Q295W, A53T_V294A_Q295W, A53T_Q161S_V294N_Q295W, A53T_Q161S_V294A, A53T_Q295W, A53T_Q161S_V294N, Q161S_V294A_Q295A, A53T_Q161S_V294A_Q295A, A53T_Q161S_Q295W, A53T_Q161S_V294N_Q295A, A53T_Q161S_Q295A, A53T_Q295A, S177W_S214R, A53T_V294A_Q295A, Q161S_V294A_Q295W_V49A_S214R, and V49A_Q295L. In some cases, said amino acid sequence is selected from the group consisting of SEQ ID NOs: 275-294; and 296-299.
In some cases, said recombinant polypeptide converts DVA and geranyl diphosphate (GPP) to CBGVA and 5-GDVA, and wherein said recombinant polypeptide produces a ratio of an amount of 5-GDVA to a combined amount of CBGVA and 5-GDVA that is higher than wild-type ORF2 enzyme under the same condition. In some cases, said ratio is 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 150%, or 200% higher than wild-type ORF2 enzyme under the same condition. In some cases, said at least one amino acid substitution to SEQ ID NO: 1 is selected from the group consisting of Q295G, Q295D, Q295S, Q295N, Q295K, and Q295R. In some cases, said amino acid sequence is selected from the group consisting of SEQ ID NOs: 140, 143, 146, 149, 153, and 153. In some cases, said at least one amino acid substitution to SEQ ID NO: 1 is selected from the group consisting of Q161V, Q161L, Q161I, Q161M, Q161W, and Q161C. In some cases, said amino acid sequence is selected from the group consisting of SEQ ID NOs: 119, 125, 127, 128, 135, and 136. In some cases, said at least one amino acid substitution to SEQ ID NO: 1 is selected from the group consisting of S214W, S214P, and S214C. In some cases, said amino acid sequence is selected from the group consisting of SEQ ID NOs: 159, 170, and 176. In some cases, said at least one amino acid substitution to SEQ ID NO: 1 is selected from the group consisting of Y288H, Y288S, Y288N, Y288K, Y288D, Y288R, and Y288E. In some cases, said amino acid sequence is selected from the group consisting of SEQ ID NOs: 100, 101, 104, 106, 109, 112, and 113.
In some cases, said recombinant polypeptide converts DVA and geranyl diphosphate (GPP) to CBGVA and 5-GDVA, and wherein said recombinant polypeptide produces a combined amount of CBGVA and 5-GDVA that is higher than wild-type ORF2 enzyme under the same condition. In some cases, said combined amount is 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 150%, or 200% higher than wild-type ORF2 enzyme under the same condition. In some cases, said at least one amino acid substitution to SEQ ID NO: 1 is selected from the group consisting of Q295D, Q295F, and Q295Y. In some cases, said amino acid sequence is selected from the group consisting of SEQ ID NOs: 140, 142, and 157. In some cases, said at least one amino acid substitution to SEQ ID NO: 1 is selected from the group consisting of Q161H, Q161C, Q161M, Q161R, Q161F, Q161T, Q161S, Q161Y, Q161L, and Q161I. In some cases, said amino acid sequence is selected from the group consisting of SEQ ID NOs: 119, 122, 124, 125, 127, 128, 132, 133, 134, and 137. In some cases, said at least one amino acid substitution to SEQ ID NO: 1 is selected from the group consisting of S214R and S214H. In some cases, said amino acid sequence is selected from the group consisting of SEQ ID NOs: 164 and 172. In some cases, said at least one amino acid substitution to SEQ ID NO: 1 is selected from the group consisting of Y288F, Y288I, Y288L, Y288M, Y288P, Y288T, and Y288W. In some cases, said amino acid sequence is selected from the group consisting of SEQ ID NOs: 102, 105, 107, 108, 110, 114, and 116.
In some cases, said at least one amino acid substitution to SEQ ID NO: 1 comprises two or more amino acid substitutions selected from the group consisting of:
(a) A17T, C25V, Q38G, V49A, V49L, V49S, A53C, A53D, A53E, A53F, A53G, A53H, A53I, A53K, A53L, A53M, A53N, A53P, A53Q, A53R, A53S, A53T, A53V, A53W, A53Y, M106E, A108G, E112D, E112G, K118N, K118Q, K119A, K119D, Y121W, F123A, F123H, F123W, Q161A, Q161C, Q161D, Q161E, Q161F, Q161G, Q161H, Q161I, Q161K, Q161L, Q161M, Q161N, Q161P, Q161R, Q161S, Q161T, Q161V, Q161W, Q161Y, M162A, M162F, D166E, N173D, L174V, S177E, S177W, S177Y, G205L, G205M, C209G, F213M, S214A, S214C, S214D, S214E, S214F, S214G, S214H, 52141, S214K, S214L, S214M, S214N, S214P, S214Q, S214R, S214T, S214V, S214W, S214Y, Y216A, L219F, D227E, R228E, R228Q, C230N, C230S, A232S, V271E, L274V, Y283L, G286E, Y288A, Y288C, Y288D, Y288E, Y288F, Y288G, Y288H, Y288I, Y288K, Y288L, Y288M, Y288N, Y288P, Y288Q, Y288R, Y288S, Y288T, Y288V, Y288W, V294A, V294F, V294N, Q295A, Q295C, Q295D, Q295E, Q295F, Q295G, Q295H, Q295I, Q295K, Q295L, Q295M, Q295N, Q295P, Q295R, Q295S, Q295T, Q295V, Q295W, Q295Y, L298A, L298Q, and L298W; or
(b) A53T_S214R, Q161S_Q295L, S177W_Q295A, Q161S_Q295F, A53T_Q295F, S214R_Q295F, Q161S_S177W, Q161S_S214R, A53T_Q161S, and A53T_V294A. In some cases, said amino acid sequence is selected from the group consisting of SEQ ID NOs: 224, 272, 287, 289, 293, 294, 296, 297, 298, and 299.
In some cases, said recombinant polypeptide converts DVA and geranyl diphosphate (GPP) to CBGVA and 5-GDVA, and wherein said recombinant polypeptide has a CBGVA production potential that is higher than wild-type ORF2 enzyme under the same condition. In some cases, said CBGVA production potential is 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 150%, or 200% higher than wild-type ORF2 enzyme under the same condition. In some cases, said at least one amino acid substitution to SEQ ID NO: 1 is selected from the group consisting of Q295F, Q295W, Q295H, Q295I, Q295M, Q295L, Q295Y, and Q295D. In some cases, said amino acid sequence is selected from the group consisting of SEQ ID NOs: 140, 142, 144, 145, 147, 148, 156, and 157. In some cases, said at least one amino acid substitution to SEQ ID NO: 1 is selected from the group consisting of Q161H, Q161R, Q161Y, Q161S, Q161F, Q161K, and Q161T. In some cases, said amino acid sequence is selected from the group consisting of SEQ ID NOs: 122, 124, 126, 132, 133, 134, and 137. In some cases, said at least one amino acid substitution to SEQ ID NO: 1 is selected from the group consisting of S214R, S214H, S214D, S214E, S214N, S214K, S214Q, and S214T. In some cases, said amino acid sequence is selected from the group consisting of SEQ ID NOs: 160, 161, 164, 166, 169, 171, 172, and 174. In some cases, said at least one amino acid substitution to SEQ ID NO: 1 is selected from the group consisting of Y288F, Y288I, Y288L, Y288M, Y288P, Y288T, Y288W, Y288C, and Y288G. In some cases, said amino acid sequence is selected from the group consisting of SEQ ID NOs: 99, 102, 103, 105, 107, 108, 110, 114, and 116.
In some cases, said at least one amino acid substitution to SEQ ID NO: 1 comprises two or more amino acid substitutions selected from the group consisting of:
(a) A17T, C25V, Q38G, V49A, V49L, V49S, A53C, A53D, A53E, A53F, A53G, A53H, A53I, A53K, A53L, A53M, A53N, A53P, A53Q, A53R, A53S, A53T, A53V, A53W, A53Y, M106E, A108G, E112D, E112G, K118N, K118Q, K119A, K119D, Y121W, F123A, F123H, F123W, Q161A, Q161C, Q161D, Q161E, Q161F, Q161G, Q161H, Q161I, Q161K, Q161L, Q161M, Q161N, Q161P, Q161R, Q161S, Q161T, Q161V, Q161W, Q161Y, M162A, M162F, D166E, N173D, L174V, S177E, S177W, S177Y, G205L, G205M, C209G, F213M, S214A, S214C, S214D, S214E, S214F, S214G, S214H, S214I, S214K, S214L, S214M, S214N, S214P, S214Q, S214R, S214T, S214V, S214W, S214Y, Y216A, L219F, D227E, R228E, R228Q, C230N, C230S, A232S, V271E, L274V, Y283L, G286E, Y288A, Y288C, Y288D, Y288E, Y288F, Y288G, Y288H, Y288I, Y288K, Y288L, Y288M, Y288N, Y288P, Y288Q, Y288R, Y288S, Y288T, Y288V, Y288W, V294A, V294F, V294N, Q295A, Q295C, Q295D, Q295E, Q295F, Q295G, Q295H, Q295I, Q295K, Q295L, Q295M, Q295N, Q295P, Q295R, Q295S, Q295T, Q295V, Q295W, Q295Y, L298A, L298Q, and L298W; or
(b) A53T_S214R, S177W_Q295A, S214R_Q295F, Q16S_S214R, S177W_S214R, Q161S_Q295L, Q161S_Q295F, V49A_S214R, A53T_Q295F, Q161S_S177W, Q161S_V294A_Q295A, A53T_Q161S_Q295W, A53T_S177W, A53T_V294A_Q295W, A53T_Q295W, Q161S_V294A_Q295W, V49A_Q295L, A53T_Q161S_V294A_Q295W A53T_Q161S_V294N_Q295A, A53T_Q161S_Q295A, A53T_Q161S_V294A_Q295A A53T_Q161S_V294N_Q295W, A53T_Q161S, A53T_Q161S_V294A, A53T_Q161S_V294N, and A53T_V294A_Q295A. In some cases, said amino acid sequence is selected from the group consisting of SEQ ID NOs: 272-274, 276-277, and 279-300.
In some cases, said recombinant polypeptide converts DVA and geranyl diphosphate (GPP) to CBGVA and 5-GDVA, and wherein said recombinant polypeptide has a 5-GDVA production potential that is higher than wild-type ORF2 enzyme under the same condition. In some cases, said 5-GDVA production potential is 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 150%, or 200% higher than wild-type ORF2 enzyme under the same condition. In some cases, said at least one amino acid substitution to SEQ ID NO: 1 is selected from the group consisting of Q295D and Q295Y. In some cases, said amino acid sequence is selected from the group consisting of SEQ ID NOs: 140 and 157. In some cases, said at least one amino acid substitution to SEQ ID NO: 1 is selected from the group consisting of Q161C, Q161M, Q161T, Q161L, Q161F, and Q161I. In some cases, said amino acid sequence is selected from the group consisting of SEQ ID NOs: 119, 122, 125, 127, 128, and 134. In some cases, said at least one amino acid substitution to SEQ ID NO: 1 comprises two or more amino acid substitutions.
In some cases, said two or more amino acid substitutions are selected from the group consisting of: A17T, C25V, Q38G, V49A, V49L, V49S, A53C, A53D, A53E, A53F, A53G, A53H, A53I, A53K, A53L, A53M, A53N, A53P, A53Q, A53R, A53S, A53T, A53V, A53W, A53Y, M106E, A108G, E112D, E112G, K118N, K118Q, K119A, K119D, Y121W, F123A, F123H, F123W, Q161A, Q161C, Q161D, Q161E, Q161F, Q161G, Q161H, Q161I, Q161K, Q161L, Q161M, Q161N, Q161P, Q161R, Q161S, Q161T, Q161V, Q161W, Q161Y, M162A, M162F, D166E, N173D, L174V, S177E, S177W, S177Y, G205L, G205M, C209G, F213M, S214A, S214C, S214D, S214E, S214F, S214G, S214H, 52141, S214K, S214L, S214M, S214N, S214P, S214Q, S214R, S214T, S214V, S214W, S214Y, Y216A, L219F, D227E, R228E, R228Q, C230N, C230S, A232S, V271E, L274V, Y283L, G286E, Y288A, Y288C, Y288D, Y288E, Y288F, Y288G, Y288H, Y288I, Y288K, Y288L, Y288M, Y288N, Y288P, Y288Q, Y288R, Y288S, Y288T, Y288V, Y288W, V294A, V294F, V294N, Q295A, Q295C, Q295D, Q295E, Q295F, Q295G, Q295H, Q295I, Q295K, Q295L, Q295M, Q295N, Q295P, Q295R, Q295S, Q295T, Q295V, Q295W, Q295Y, L298A, L298Q, and L298W.
In some cases, said two or more amino acid substitutions are selected from the group consisting of: Q161S_Q295F, A53T_Q295F, S214R_Q295F, Q161S_S177W, Q16S_S214R, A53T_S177W, A53T_V294A_Q295W, A53T_Q161S_V294A_Q295W, A53T_Q295W, Q161S_V294A_Q295A, A53T_Q161S_V294N_Q295W, A53T_Q161S, A53T_Q161S_Q295W, S177W_S214R, A53T_Q161S_V294A_Q295A, A53T_Q161S_V294N_Q295A, A53T_Q161S_Q295A, A53T_Q161S_V294A, Q161S_V294A_Q295W, and A53T_Q161S_V294N. In some cases, said amino acid sequence is selected from the group consisting of SEQ ID NOs: 273, 274, 276, 277, 279-290, and 293-300.
In some cases, said recombinant polypeptide converts Olivetol (O) and geranyl diphosphate (GPP) to CBG and 5-GO, and wherein said recombinant polypeptide produces a ratio of an amount of CBG to a combined amount of CBG and 5-GO that is higher than wild-type ORF2 enzyme under the same condition. In some cases, said ratio is 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 150%, or 200% higher than wild-type ORF2 enzyme under the same condition. In some cases, said at least one amino acid substitution to SEQ ID NO: 1 is selected from the group consisting of Q295G, Q295M, Q295H, Q295F, Q295C, Q295L, and Q295S. In some cases, said amino acid sequence is selected from the group consisting of SEQ ID NOs: 139, 142, 143, 144, 147, 148, and 153. In some cases, said at least one amino acid substitution to SEQ ID NO: 1 is selected from the group consisting of Q161F, Q161Y, Q161L, Q161H, Q161M, Q161I, Q161C, Q161S, Q161T, Q161D, Q161G, Q161W, and Q161N. In some cases, said amino acid sequence is selected from the group consisting of SEQ ID NOs: 119, 120, 122, 123, 124, 125, 127, 128, 129, 133, 134, 136, and 137. In some cases, said at least one amino acid substitution to SEQ ID NO: 1 is selected from the group consisting of S214Q, S214H, S214E, S214N, S214F, S214L, S214I, S214M, S214K, and S214D. In some cases, said amino acid sequence is selected from the group consisting of SEQ ID NOs: 160, 161, 162, 164, 165, 166, 167, 168, 169, and 171. In some cases, said at least one amino acid substitution to SEQ ID NO: 1 is selected from the group consisting of Y288F, Y288L, Y288I, Y288M, Y288T, Y288C, Y288A, Y288W, Y288P, Y288S, Y288G, and Y288R. In some cases, said amino acid sequence is selected from the group consisting of SEQ ID NOs: 98, 99, 102, 103, 105, 107, 108, and 109-117.
In some cases, said at least one amino acid substitution to SEQ ID NO: 1 comprises two or more amino acid substitution to SEQ ID NO: 1 selected from the group consisting of:
(a) A17T, C25V, Q38G, V49A, V49L, V49S, A53C, A53D, A53E, A53F, A53G, A53H, A53I, A53K, A53L, A53M, A53N, A53P, A53Q, A53R, A53S, A53T, A53V, A53W, A53Y, M106E, A108G, E112D, E112G, K118N, K118Q, K119A, K119D, Y121W, F123A, F123H, F123W, Q161A, Q161C, Q161D, Q161E, Q161F, Q161G, Q161H, Q161I, Q161K, Q161L, Q161M, Q161N, Q161P, Q161R, Q161S, Q161T, Q161V, Q161W, Q161Y, M162A, M162F, D166E, N173D, L174V, S177E, S177W, S177Y, G205L, G205M, C209G, F213M, S214A, S214C, S214D, S214E, S214F, S214G, S214H, 52141, S214K, 5214L, S214M, S214N, S214P, S214Q, S214R, S214T, S214V, S214W, S214Y, Y216A, L219F, D227E, R228E, R228Q, C230N, C230S, A232S, V271E, L274V, Y283L, G286E, Y288A, Y288C, Y288D, Y288E, Y288F, Y288G, Y288H, Y288I, Y288K, Y288L, Y288M, Y288N, Y288P, Y288Q, Y288R, Y288S, Y288T, Y288V, Y288W, V294A, V294F, V294N, Q295A, Q295C, Q295D, Q295E, Q295F, Q295G, Q295H, Q295I, Q295K, Q295L, Q295M, Q295N, Q295P, Q295R, Q295S, Q295T, Q295V, Q295W, Q295Y, L298A, L298Q, and L298W; or
(b) A53T, Q161S, V294A, and Q295A; A53T, Q161S, and Q295A; A53T, Q161S, and V294A; A53T, Q161S, V294N, and Q295A; A53T, Q161S, and V294N; Q161S, V294A, and Q295A; A53T and V294A; A53T, Q161S, and Q295W; A53T, V294A, and Q295A; A53T, Q161S, V294N, and Q295W; A53T, Q161S, V294A, and Q295W; Q161S and Q295L; A53T and Q161S; Q161S, V294A, and Q295W; Q161S and Q295F; A53T and Q295W; A53T and Q295A; A53T, V294A, and Q295W; A53T and Q295F; A53T and S177W; S177W and Q295A; Q161S and S177W; S214R and Q295F; A53T and 5214R; Q165 and 5214R; Q161S and Q295L; A53T and Q161S; Q161S and Q295F; A53T and Q295F; and A53T and V294A. In some cases, said amino acid sequence is selected from the group consisting of SEQ ID NOs: 275-294; and 296-299.
In some cases, said recombinant polypeptide converts O and geranyl diphosphate (GPP) to CBG and 5-GO, and wherein said recombinant polypeptide produces a ratio of an amount of 5-GO to a combined amount of CBG and 5-GO that is higher than wild-type ORF2 enzyme under the same condition. In some cases, said ratio is 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 150%, or 200% higher than wild-type ORF2 enzyme under the same condition. In some cases, said at least one amino acid substitution to SEQ ID NO: 1 is selected from the group consisting of Q295Y, Q295N, Q295D, Q295W, Q295T, Q295V, Q295P, Q295I, Q295R, Q295K, and Q295E. In some cases, said amino acid sequence is selected from the group consisting of SEQ ID NOs: 140, 141, 145, 146, 149, 150, 152, 154, 155, 156, and 157. In some cases, said at least one amino acid substitution to SEQ ID NO: 1 is selected from the group consisting of Q161R, Q161K, and Q161P. In some cases, said amino acid sequence is selected from the group consisting of SEQ ID NOs: 126, 130, and 132. In some cases, said at least one amino acid substitution to SEQ ID NO: 1 is selected from the group consisting of S214W, S214V, S214T, S214R, S214C, S214P, and S214G. In some cases, said amino acid sequence is selected from the group consisting of SEQ ID NOs: 159, 163, 170, 172, 174, 175, and 176. In some cases, said at least one amino acid substitution to SEQ ID NO: 1 is selected from the group consisting of Y288N, Y288H, Y288D, Y288K, and Y288E. In some cases, said amino acid sequence is selected from the group consisting of SEQ ID NOs: 100, 101, 104, 106, and 109. In some cases, said at least one amino acid substitution to SEQ ID NO: 1 is selected from the group consisting of V49A and Q295L; V49A and S214R; and S177W and S214R. In some cases, said amino acid sequence is selected from the group consisting of SEQ ID NOs: 288, 291, and 292.
In some cases, said recombinant polypeptide converts O and geranyl diphosphate (GPP) to CBG and 5-GO, and wherein said recombinant polypeptide produces a combined amount of CBG and 5-GO that is higher than wild-type ORF2 enzyme under the same condition. In some cases, said combined amount is 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 150%, or 200% higher than wild-type ORF2 enzyme under the same condition. In some cases, said at least one amino acid substitution to SEQ ID NO: 1 is selected from the group consisting of Q295H, Q295M, Q295G, Q295F, Q295W, and Q295D. In some cases, said amino acid sequence is selected from the group consisting of SEQ ID NOs: 140, 142, 143, 144, 148, and 156. In some cases, said at least one amino acid substitution to SEQ ID NO: 1 is selected from the group consisting of Q161F, Q161Y, Q161L, Q161H, Q161I, Q161M, Q161C, Q161T, Q161S, Q161D, Q161G, Q161R, Q161N, and Q161W. In some cases, said amino acid sequence is selected from the group consisting of SEQ ID NOs: 119, 120, 122, 123, 124, 125, 127, 128, 129, 132, 133, 134, 136, and 137. In some cases, said at least one amino acid substitution to SEQ ID NO: 1 is selected from the group consisting of S214Q, S214H, S214E, S214N, S214F, S214I, S214L, and S214M. In some cases, said amino acid sequence is selected from the group consisting of SEQ ID NOs: 161, 162, 164, 165, 167, 168, 169, and 171. In some cases, said at least one amino acid substitution to SEQ ID NO: 1 is selected from the group consisting of Y288F, Y288L, Y288I, Y288M, Y288T, Y288C, and Y288P. In some cases, said amino acid sequence is selected from the group consisting of SEQ ID NOs: 99, 102, 105, 107, 108, 110, and 114.
In some cases, said at least one amino acid substitution to SEQ ID NO: 1 comprises two or more amino acid substitutions selected from the group consisting of:
(a) A17T, C25V, Q38G, V49A, V49L, V49S, A53C, A53D, A53E, A53F, A53G, A53H, A53I, A53K, A53L, A53M, A53N, A53P, A53Q, A53R, A53S, A53T, A53V, A53W, A53Y, M106E, A108G, E112D, E112G, K118N, K118Q, K119A, K119D, Y121W, F123A, F123H, F123W, Q161A, Q161C, Q161D, Q161E, Q161F, Q161G, Q161H, Q161I, Q161K, Q161L, Q161M, Q161N, Q161P, Q161R, Q161S, Q161T, Q161V, Q161W, Q161Y, M162A, M162F, D166E, N173D, L174V, S177E, S177W, S177Y, G205L, G205M, C209G, F213M, S214A, S214C, S214D, S214E, S214F, S214G, S214H, S214I, S214K, S214L, S214M, S214N, S214P, S214Q, S214R, S214T, S214V, S214W, S214Y, Y216A, L219F, D227E, R228E, R228Q, C230N, C230S, A232S, V271E, L274V, Y283L, G286E, Y288A, Y288C, Y288D, Y288E, Y288F, Y288G, Y288H, Y288I, Y288K, Y288L, Y288M, Y288N, Y288P, Y288Q, Y288R, Y288S, Y288T, Y288V, Y288W, V294A, V294F, V294N, Q295A, Q295C, Q295D, Q295E, Q295F, Q295G, Q295H, Q295I, Q295K, Q295L, Q295M, Q295N, Q295P, Q295R, Q295S, Q295T, Q295V, Q295W, Q295Y, L298A, L298Q, and L298W; or
(b) Q161S and Q295L; A53T and Q161S; Q161S and Q295F; A53T, Q161S, V294A, and Q295A; A53T, Q161S, and Q295A; A53T and Q295F; A53T, Q161S, and V294A; A53T, Q161S, V294N, and Q295A; A53T, Q161S, and V294N; Q161S, V294A, and Q295A; A53T and V294A; A53T, Q161S, and Q295W; A53T, V294A, and Q295A; A53T, Q161S, V294N, and Q295W; A53T, Q161S, V294A, and Q295W; Q161S, V294A, and Q295W; A53T and S177W; A53T and Q295W; A53T and Q295W; A53T and Q295A; A53T, V294A, and Q295W; S177W and Q295A; and Q161S and S177W.
In some cases, said amino acid sequence is selected from the group consisting of SEQ ID NOs: 214, 224, 272, 273, 274, 275, 276, 277, 278, 279, 280, 281, 282, 283, 284, 285, 286, 287, 289, 290, 293, 296, and 297. In some cases, said recombinant polypeptide converts O and geranyl diphosphate (GPP) to CBG and 5-GO, and wherein said recombinant polypeptide has a CBG production potential that is higher than wild-type ORF2 enzyme under the same condition. In some cases, said CBGVA production potential is 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 150%, or 200% higher than wild-type ORF2 enzyme under the same condition. In some cases, said at least one amino acid substitution to SEQ ID NO: 1 is selected from the group consisting of Q295H, Q295M, Q295G, Q295F, Q295W, and Q295C. In some cases, said amino acid sequence is selected from the group consisting of SEQ ID NOs: 139, 142, 143, 144, 148, and 156. In some cases, said at least one amino acid substitution to SEQ ID NO: 1 is selected from the group consisting of Q161F, Q161Y, Q161L, Q161H, Q161M, Q161I, Q161C, Q161S, Q161T, Q161D, Q161G, and Q161W. In some cases, said amino acid sequence is selected from the group consisting of SEQ ID NOs: 119, 120, 122, 123, 124, 125, 127, 128, 133, 134, 136, and 137. In some cases, said at least one amino acid substitution to SEQ ID NO: 1 is selected from the group consisting of S214Q, S214H, S214E, S214N, S214F, 52141, S214L, and S214M. In some cases, said amino acid sequence is selected from the group consisting of SEQ ID NOs: 161, 162, 164, 165, 167, 168, 169, and 171. In some cases, said at least one amino acid substitution to SEQ ID NO: 1 is selected from the group consisting of Y288F, Y288L, Y288I, Y288M, Y288T, Y288C, and Y288W. In some cases, said amino acid sequence is selected from the group consisting of SEQ ID NOs: 99, 102, 105, 107, 108, 114, and 116.
In some cases, said at least one amino acid substitution to SEQ ID NO: 1 comprises two or more amino acid substitutions selected from the group consisting of:
(a) A17T, C25V, Q38G, V49A, V49L, V49S, A53C, A53D, A53E, A53F, A53G, A53H, A53I, A53K, A53L, A53M, A53N, A53P, A53Q, A53R, A53S, A53T, A53V, A53W, A53Y, M106E, A108G, E112D, E112G, K118N, K118Q, K119A, K119D, Y121W, F123A, F123H, F123W, Q161A, Q161C, Q161D, Q161E, Q161F, Q161G, Q161H, Q161I, Q161K, Q161L, Q161M, Q161N, Q161P, Q161R, Q161S, Q161T, Q161V, Q161W, Q161Y, M162A, M162F, D166E, N173D, L174V, S177E, S177W, S177Y, G205L, G205M, C209G, F213M, S214A, S214C, S214D, S214E, S214F, S214G, S214H, 52141, S214K, S214L, S214M, S214N, S214P, S214Q, S214R, S214T, S214V, S214W, S214Y, Y216A, L219F, D227E, R228E, R228Q, C230N, C230S, A232S, V271E, L274V, Y283L, G286E, Y288A, Y288C, Y288D, Y288E, Y288F, Y288G, Y288H, Y288I, Y288K, Y288L, Y288M, Y288N, Y288P, Y288Q, Y288R, Y288S, Y288T, Y288V, Y288W, V294A, V294F, V294N, Q295A, Q295C, Q295D, Q295E, Q295F, Q295G, Q295H, Q295I, Q295K, Q295L, Q295M, Q295N, Q295P, Q295R, Q295S, Q295T, Q295V, Q295W, Q295Y, L298A, L298Q, and L298W; or
(b) A53T_Q161S_V294A_Q295W, A53T_Q161S_V294A, Q161S_Q295L, A53T_Q295F, A53T_Q295A, A53T_Q161S_V294N, Q161S_Q295F, A53T_Q161S_Q295W, A53T_Q161S, A53T_S177W, Q161S_V294A_Q295W, A53T_V294A, A53T_V294A_Q295W, S177W_Q295A, Q161S_S177W, and A53T_S214R.
In some cases, said amino acid sequence is selected from the group consisting of SEQ ID NOs: 224, 272, 273, 275, 280, 282, 283, 284, 285, 287, 289, 290, 293, 294, 296, and 297. In some cases, said recombinant polypeptide converts O and geranyl diphosphate (GPP) to CBG and 5-GO, and wherein said recombinant polypeptide has a 5-GO production potential that is higher than wild-type ORF2 enzyme under the same condition. In some cases, said 5-GO production potential is 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 150%, or 200% higher than wild-type ORF2 enzyme under the same condition. In some cases, said at least one amino acid substitution to SEQ ID NO: 1 is selected from the group consisting of Q295H, Q295W, and Q295F. In some cases, said amino acid sequence is selected from the group consisting of SEQ ID NOs: 142, 144, and 156. In some cases, said at least one amino acid substitution to SEQ ID NO: 1 is selected from the group consisting of Q161L, Q161Y, Q161F, Q161I, Q161C, Q161M, Q161T, Q161H, Q161S, Q161R, Q161G, Q161N, and Q161D. In some cases, said amino acid sequence is selected from the group consisting of SEQ ID NOs: 119, 120, 122, 123, 124, 125, 127, 128, 129, 132, 133, 134, and 137. In some cases, said at least one amino acid substitution to SEQ ID NO: 1 is S214E, 52141, S214T, S214F, S214M, S214L, S214G, S214N, S214V, S214C, S214D, and S214Y. In some cases, said amino acid sequence is SEQ ID NO: 159, 160, 161, 162, 163, 165, 167, 168, 169, 174, 175, and 177. In some cases, said at least one amino acid substitution to SEQ ID NO: 1 is Y288F and Y288L. In some cases, said amino acid sequence is SEQ ID NO: 102 and 107.
In another aspect, the disclosure provides recombinant polypeptide comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 1, wherein said recombinant polypeptide converts olivetolic acid (OA) and geranyl diphosphate (GPP) to cannabigerolic acid (CBGA) and 5-geranyl olivetolate (5-GOA), and wherein said recombinant polypeptide produces a ratio of an amount of CBGA to a combined amount of CBGA and 5-GOA that is higher than wild-type ORF2 enzyme under the same condition.
In one aspect, the disclosure provides a recombinant polypeptide comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 1, wherein said recombinant polypeptide converts olivetolic acid (OA) and geranyl diphosphate (GPP) to cannabigerolic acid (CBGA) and 5-geranyl olivetolate (5-GOA), and wherein said recombinant polypeptide produces a ratio of an amount of 5-GOA to a combined amount of CBGA and 5-GOA that is higher than wild-type ORF2 enzyme under the same condition.
In one aspect, the disclosure provides a recombinant polypeptide comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 1, wherein said recombinant polypeptide converts olivetolic acid (OA) and geranyl diphosphate (GPP) to cannabigerolic acid (CBGA) and 5-geranyl olivetolate (5-GOA), and wherein said recombinant polypeptide produces a combined amount of CBGA and 5-GOA that is higher than wild-type ORF2 enzyme under the same condition.
In one aspect, the disclosure provides a recombinant polypeptide comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 1, wherein said recombinant polypeptide converts olivetolic acid (OA) and geranyl diphosphate (GPP) to cannabigerolic acid (CBGA) and 5-geranyl olivetolate (5-GOA), and wherein said recombinant polypeptide has a CBGA production potential that is higher than wild-type ORF2 enzyme under the same condition.
In one aspect, the disclosure provides a recombinant polypeptide comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 1, wherein said recombinant polypeptide converts olivetolic acid (OA) and geranyl diphosphate (GPP) to cannabigerolic acid (CBGA) and 5-geranyl olivetolate (5-GOA), and wherein said recombinant polypeptide has a 5-GOA production potential that is higher than wild-type ORF2 enzyme under the same condition.
In one aspect, the disclosure provides a recombinant polypeptide comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 1, wherein said recombinant polypeptide converts DVA and geranyl diphosphate (GPP) to CBGVA and 5-GDVA, and wherein said recombinant polypeptide produces a ratio of an amount of CBGVA to a combined amount of CBGVA and 5-GDVA that is higher than wild-type ORF2 enzyme under the same condition.
In another aspect, the disclosure provides a recombinant polypeptide comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 1, wherein said recombinant polypeptide converts DVA and geranyl diphosphate (GPP) to CBGVA and 5-GDVA, and wherein said recombinant polypeptide produces a ratio of an amount of 5-GDVA to a combined amount of CBGVA and 5-GDVA that is higher than wild-type ORF2 enzyme under the same condition.
In one aspect, the disclosure provides a recombinant polypeptide comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 1, wherein said recombinant polypeptide converts DVA and geranyl diphosphate (GPP) to CBGVA and 5-GDVA, and wherein said recombinant polypeptide produces a combined amount of CBGVA and 5-GDVA that is higher than wild-type ORF2 enzyme under the same condition.
In another aspect, the disclosure provides a recombinant polypeptide comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 1, wherein said recombinant polypeptide converts DVA and geranyl diphosphate (GPP) to CBGVA and 5-GDVA, and wherein said recombinant polypeptide has a CBGVA production potential that is higher than wild-type ORF2 enzyme under the same condition.
In one aspect, the disclosure provides a recombinant polypeptide comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 1, wherein said recombinant polypeptide converts DVA and geranyl diphosphate (GPP) to CBGVA and 5-GDVA, and wherein said recombinant polypeptide has a 5-GDVA production potential that is higher than wild-type ORF2 enzyme under the same condition.
In another aspect, the disclosure provides a recombinant polypeptide comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 1, wherein said recombinant polypeptide converts Olivetol (O) and geranyl diphosphate (GPP) to CBG and 5-GO, and wherein said recombinant polypeptide produces a ratio of an amount of CBG to a combined amount of CBG and 5-GO that is higher than wild-type ORF2 enzyme under the same condition.
In one aspect, the disclosure provides a recombinant polypeptide comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 1, wherein said recombinant polypeptide converts Olivetol (O) and geranyl diphosphate (GPP) to CBG and 5-GO, and wherein said recombinant polypeptide produces a ratio of an amount of 5-GO to a combined amount of CBG and 5-GO that is higher than wild-type ORF2 enzyme under the same condition.
In one aspect, the disclosure provides a recombinant polypeptide comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 1, wherein said recombinant polypeptide converts Olivetol (O) and geranyl diphosphate (GPP) to CBG and 5-GO, and wherein said recombinant polypeptide produces a combined amount of CBG and 5-GO that is higher than wild-type ORF2 enzyme under the same condition.
In one aspect, the disclosure provides a recombinant polypeptide comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 1, wherein said recombinant polypeptide converts Olivetol (O) and geranyl diphosphate (GPP) to CBG and 5-GO, and wherein said recombinant polypeptide has a CBG production potential that is higher than wild-type ORF2 enzyme under the same condition.
In one aspect, the disclosure provides a recombinant polypeptide comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 1, wherein said recombinant polypeptide converts Olivetol (O) and geranyl diphosphate (GPP) to CBG and 5-GO, and wherein said recombinant polypeptide has a 5-GO production potential that is higher than wild-type ORF2 enzyme under the same condition.
In some cases, said amino acid sequence has at least 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 1. In some cases, said at least one amino acid substitution comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 amino acid substitutions to SEQ ID NO: 1. In some cases, said at least one amino acid substitution is located on a position chosen from the group consisting of 1-46, 48-63, 65-109, 111-120, 122, 124, 125, 127-160, 162-174, 176, 178-213, 215, 217-287, 289-293, and 296-307. In some cases, said amino acid sequence comprises said at least one amino acid substitution to SEQ ID NO: 1 at a position selected from the group consisting of A17, C25, Q38, V49, A53, M106, A108, E112, K118, K119, Y121, F123, Q161, M162, D166, N173, L174, S177, G205, C209, F213, S214, Y216, L219, D227, R228, C230, A232, V271, L274, Y283, G286, Y288, V294, Q295, and L298. In some cases, said at least one amino acid substitution to SEQ ID NO: 1 is selected from the group consisting of A17T, C25V, Q38G, V49A, V49L, V49S, A53C, A53D, A53E, A53F, A53G, A53H, A53I, A53K, A53L, A53M, A53N, A53P, A53Q, A53R, A53S, A53T, A53V, A53W, A53Y, M106E, A108G, E112D, E112G, K118N, K118Q, K119A, K119D, Y121W, F123A, F123H, F123W, Q161A, Q161C, Q161D, Q161E, Q161F, Q161G, Q161H, Q161I, Q161K, Q161L, Q161M, Q161N, Q161P, Q161R, Q161S, Q161T, Q161V, Q161W, Q161Y, M162A, M162F, D166E, N173D, L174V, S177E, S177W, S177Y, G205L, G205M, C209G, F213M, S214A, S214C, S214D, S214E, S214F, S214G, S214H, S214I, S214K, S214L, S214M, S214N, S214P, S214Q, S214R, S214T, S214V, S214W, S214Y, Y216A, L219F, D227E, R228E, R228Q, C230N, C230S, A232S, V271E, L274V, Y283L, G286E, Y288A, Y288C, Y288D, Y288E, Y288F, Y288G, Y288H, Y288I, Y288K, Y288L, Y288M, Y288N, Y288P, Y288Q, Y288R, Y288S, Y288T, Y288V, Y288W, V294A, V294F, V294N, Q295A, Q295C, Q295D, Q295E, Q295F, Q295G, Q295H, Q295I, Q295K, Q295L, Q295M, Q295N, Q295P, Q295R, Q295S, Q295T, Q295V, Q295W, Q295Y, L298A, L298Q, and L298W. In some cases, the recombinant polypeptide comprises any one of the recombinant polypeptides disclosed herein.
In some aspects, the disclosure provides a nucleic acid molecule, comprising a nucleotide sequence encoding any one of the recombinant polypeptides disclosed herein, or a codon degenerate nucleotide sequence thereof.
In one aspect, the disclosure provides a nucleic acid molecule, comprising a nucleotide sequence with at least 95% sequence identity to a sequence selected from the group consisting of SEQ ID NO: 1-198 or a codon degenerate nucleotide sequence thereof. In some cases, the nucleotide sequence comprises at least 500, 600, 700, 800, or 900 nucleotides. In some cases, said nucleic acid molecule is isolated and purified.
In one aspect, the disclosure provides a cell vector, construct or expression system comprising any or more of the nucleic acid molecules disclosed herein.
In one aspect, the disclosure provides a cell, comprising said cell vector, construct or expression system disclosed herein. In some cases, said cell is a bacteria, yeast, insect, mammalian, fungi, vascular plant, or non-vascular plant cell. In some cases, said cell is a microalgae cell. In some cases, said cell is an E. coli cell.
In one aspect, the disclosure provides a plant comprising any cell disclosed herein. In some cases, the plant is a terrestrial plant.
In one aspect, the disclosure provides a method of producing a prenylation product, comprising: contacting said recombinant polypeptide of any one of claims 1-198 with a substrate and a donor, thereby producing said prenylation product. In some cases, said substrate comprises olivetolic acid (OA), divarinolic acid (DVA), olivetol (O), resveratrol, piceattanol and related stilbenes, naringenin, apigenin and related flavanones and flavones, respectively, Isoliquiritigenin, 2′-O-methylisoliquiritigenin and related chalcones, catechins and epi-catechins of all possible stereoisomers, biphenyl compounds such as 3,5-dihydroxy-biphenyl, benzophenones such as phlorobenzophenone, isoflavones such as biochanin A, genistein, daidzein, 2,4-dihydroxybenzoic acid, 1,3-benzenediol, 2,4-dihydroxy-6-methylbenzoic acid; 1,3-Dihydroxy-5-methylbenzene; 2,4-Dihydroxy-6-aethyl-benzoesaeure; 5-ethylbenzene-1,3-diol 2,4-dihydroxy-6-propylbenzoic acid; 5-propylbenzene-1,3-diol; 2-butyl-4,6-dihydroxybenzoic acid; 5-butylbenzene-1,3-diol; 2,4-dihydroxy-6-pentyl-benzoic acid; 5-pentylbenzene-1,3-diol; 5-hexylbenzene-1,3-diol; 2-heptyl-4,6-dihydroxy-benzoic acid; 5-heptylbenzene-1,3-diol; 5-Dodecylbenzene-1,3-diol; 5-nonadecylbenzene-1,3-diol; 1,3-Benzenediol; 3,4′,5-Trihydroxystilbene; 4′5-Tetrahydroxystilbene; 1,2-Diphenylethylene; 2-Phenylbenzopyran-4-one; 2-Phenylchroman-4-one; 1,3-benzenediol; 5,7,4′-Trihydroxyflavone; (E)-1-(2,4-dihydroxyphenyl)-3-(4-hydroxyphenyl)prop-2-en-1-one; 4,4′-dihydroxy-2′-methoxychalcone; 1,3-Diphenylpropenone; (2R,3 S)-2-(3,4-Dihydroxyphenyl)chroman-3,5,7-triol; (2R,3R)-2-(3,4-Dihydroxyphenyl)-3,5,7-chromanetriol; Phenylbenzene; 5-Phenylresorcinol; diphenylmethanone; 3-phenyl-4H-chromen-4-one; 5,7-Dihydroxy-3-(4-methoxyphenyl)-4H-chromen-4-one; 4′,5,7-Trihydroxyisoflavone; 4′,7-Dihydroxyisoflavone; 4-Hydroxy-6-methyl-2H-pyran-2-one; 1,6-DHN; or any combination thereof. In some cases, said substrate comprises olivetolic acid (OA), olivetol (O), or divarinic acid (DVA). In some cases, said donor comprises dimethylallyl pyrophosphate (DMAPP), geranyl pyrophosphate (GPP), or farnesyl pyrophosphate (FPP). In some cases, said substrate comprises olivetolic acid (OA) and said donor comprises geranyl pyrophosphate (GPP), and wherein said prenylation product comprises CBGA, 5-GOA, or a prenylated product on any other position on an aromatic ring of said substrate.
In some cases, said prenylation product comprises CBGA and 5-GOA, and wherein said recombinant polypeptide produces a ratio of an amount of CBGA to a combined amount of CBGA and 5-GOA that is higher than wild-type ORF2 enzyme under the same acid condition. In some cases, said prenylation product comprises CBGA and 5-GOA, and wherein said recombinant polypeptide produces a ratio of an amount of 5-GOA to a combined amount of CBGA and 5-GOA that is higher than wild-type ORF2 enzyme under the same acid condition. In some cases, said prenylation product comprises CBGA and 5-GOA, and wherein said recombinant polypeptide produces a combined amount of CBGA and 5-GOA that is higher than wild-type ORF2 enzyme under the same acid condition. In some cases, said prenylation product comprises CBGA and 5-GOA, and wherein said recombinant polypeptide has a CBGA production potential that is higher than wild-type ORF2 enzyme under the same acid condition.
In some cases, said acid condition has a pH of less than 7. In some cases, the method of claim 219, wherein said acid condition has a pH of less than 6. In some cases, said acid condition has a pH of about 5.3. In some cases, said substrate comprises olivetol (O) and said donor comprises geranyl pyrophosphate (GPP), and wherein said prenylation product comprises CBG, 5-GO or a prenylated product on any other position on an aromatic ring of said substrate. In some cases, said substrate comprises divarinic acid (DVA) and said donor comprises geranyl pyrophosphate (GPP), and wherein said prenylation product comprises CBGVA, 5-GDVA or a prenylated product on any other position on an aromatic ring of said substrate.
In some cases, said contacting comprises contacting said substrate and said donor with said recombinant polypeptide in a cell. In some cases, said cell is a bacteria, yeast, insect, mammalian, fungi, microalgae, or plant cell. In some cases, said contacting comprises contacting said substrate and said donor with said recombinant polypeptide in a cell-free reaction. In some cases, said contacting comprises contacting said substrate and said donor with said recombinant polypeptide in a reaction, wherein said recombinant polypeptide is secreted by a cell into a media before the contacting.
In yet another aspect, the disclosure provides a composition, comprising a prenylation product and any one recombinant polypeptide disclosed herein. In some cases, said prenylation product comprises CBGA, CBG, CBGVA or any combination thereof. In some cases, said prenylation product comprises 5-GOA, 5-GO. In some cases, said prenylation product comprises 5-GDVA.
In one aspect, the disclosure provides a method of screening for an ORF2 mutant, comprising: providing two or more ORF2 mutants; contacting each of said two or more ORF2 mutants with a substrate and a donor, thereby producing at least two prenylation products under the same condition; measuring a ratio of an amount of one of said at least two prenylation products to a combined amount of said at least two prenylation products for each of said two or more ORF2 mutants; and identifying said ORF2 mutant with the highest ratio among said two or more ORF2 mutants. In some cases, each of said two or more ORF2 mutants comprises an amino acid sequence with at least 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 1. In some cases, said two or more ORF2 mutants comprises different amino acid sequences. In some cases, each of said amino acid sequences comprises at least one amino acid substitution to SEQ ID NO: 1. In some cases, each of said at least one amino acid substitution is located at a different amino acid position. In some cases, said at least one amino acid substitution is located on a position chosen from the group consisting of amino acid positions 1-46, 48-63, 65-109, 111-120, 122, 124, 125, 127-160, 162-174, 176, 178-213, 215, 217-287, 289-293, and 296-307. In some cases, said at least one amino acid substitution is located on a position chosen from the group consisting of amino acid positions 17, 25, 38, 49, 53, 106, 108, 112, 118, 119, 121, 123, 161, 162, 166, 173, 174, 177, 205, 209, 213, 214, 216, 219, 227, 228, 230, 232, 271, 274, 283, 286, 288, 294, 295, and 298.
In some cases, said substrate comprises olivetolic acid (OA), divarinolic acid (DVA), olivetol (O), resveratrol, piceattanol and related stilbenes, naringenin, apigenin and related flavanones and flavones, respectively, Isoliquiritigenin, 2′-O-methylisoliquiritigenin and related chalcones, catechins and epi-catechins of all possible stereoisomers, biphenyl compounds such as 3,5-dihydroxy-biphenyl, benzophenones such as phlorobenzophenone, isoflavones such as biochanin A, genistein, daidzein, 2,4-dihydroxybenzoic acid, 1,3-benzenediol, 2,4-dihydroxy-6-methylbenzoic acid; 1,3-Dihydroxy-5-methylbenzene; 2,4-Dihydroxy-6-aethyl-benzoesaeure; 5-ethylbenzene-1,3-diol 2,4-dihydroxy-6-propylbenzoic acid; 5-propylbenzene-1,3-diol; 2-butyl-4,6-dihydroxybenzoic acid; 5-butylbenzene-1,3-diol; 2,4-dihydroxy-6-pentyl-benzoic acid; 5-pentylbenzene-1,3-diol; 5-hexylbenzene-1,3-diol; 2-heptyl-4,6-dihydroxy-benzoic acid; 5-heptylbenzene-1,3-diol; 5-Dodecylbenzene-1,3-diol; 5-nonadecylbenzene-1,3-diol; 1,3-Benzenediol; 3,4′,5-Trihydroxystilbene; 4'S-Tetrahydroxystilbene; 1,2-Diphenylethylene; 2-Phenylbenzopyran-4-one; 2-Phenylchroman-4-one; 1,3-benzenediol; 5,7,4′-Trihydroxyflavone; (E)-1-(2,4-dihydroxyphenyl)-3-(4-hydroxyphenyl)prop-2-en-1-one; 4,4′-dihydroxy-2′-methoxychalcone; 1,3-Diphenylpropenone; (2R,3 S)-2-(3,4-Dihydroxyphenyl)chroman-3,5,7-triol; (2R,3R)-2-(3,4-Dihydroxyphenyl)-3,5,7-chromanetriol; Phenylbenzene; 5-Phenylresorcinol; diphenylmethanone; 3-phenyl-4H-chromen-4-one; 5,7-Dihydroxy-3-(4-methoxyphenyl)-4H-chromen-4-one; 4′,5,7-Trihydroxyisoflavone; 4′,7-Dihydroxyisoflavone; 4-Hydroxy-6-methyl-2H-pyran-2-one; 1,6-DHN; or any combination thereof.
In some cases, said substrate comprises olivetolic acid (OA), olivetol (O), or divarinic acid (DVA). In some cases, said donor comprises dimethylallyl pyrophosphate (DMAPP), geranyl pyrophosphate (GPP), or farnesyl pyrophosphate (FPP). In some cases, said substrate comprises olivetolic acid (OA) and said donor comprises geranyl pyrophosphate (GPP), and wherein said at least two prenylation products comprises CBGA and 5-GOA. In some cases, said substrate comprises olivetol (O) and said donor comprises geranyl pyrophosphate (GPP), and wherein said at least two prenylation products comprises CBG and 5-GO. In some cases, said substrate comprises divarinic acid (DVA) and said donor comprises geranyl pyrophosphate (GPP), and said at least two prenylation products comprises CBGVA and 5-GDVA.
In one aspect, the disclosure provides methods of making a prenyltransferase mutant, comprising: aligning a crystal structure of a prenyltransferase with a crystal structure of wild-type ORF2 enzyme; identifying an amino acid position on said prenyltransferase corresponding to any one of amino acid positions 17, 25, 38, 49, 53, 106, 108, 112, 118, 119, 162, 166, 173, 174, 205, 209, 213, 219, 227, 228, 230, 232, 271, 274, 283, 286, 288, and 298 on said wild-type ORF2 enzyme, based on alignment of said crystal structures; and substitute said amino acid position on said prenyltransferase with a different amino acid, thereby generating said prenyltransferase mutant.
In one aspect, the disclosure provides a method of making a prenyltransferase mutant, comprising: aligning an amino acid sequence of a prenyltransferase with an amino acid sequence of wild-type ORF2 enzyme; identifying an amino acid position on said prenyltransferase corresponding to any one of amino acid positions 17, 25, 38, 49, 53, 106, 108, 112, 118, 119, 162, 166, 173, 174, 205, 209, 213, 219, 227, 228, 230, 232, 271, 274, 283, 286, 288, and 298 on said wild-type ORF2 enzyme, based on alignment of said amino acid sequences; and substitute said amino acid position on said prenyltransferase with a different amino acid, thereby generating said prenyltransferase mutant.
In some cases, said at least one amino acid substitution to SEQ ID NO: 1 is selected from the group consisting of A53T, A53I, A53V, A53L, A53M, A53K, A53S, A53C, and A53Q. In some cases, said amino acid sequence is selected from the group consisting of SEQ ID NOs: 179, 185, 186, 187, 188, 191, 193, 194, and 195. In some cases, said at least one amino acid substitution to SEQ ID NO: 1 is selected from the group consisting of A53L, A53K, A53V, A53M, A53Q, A53T, A53S, A53C, and A53I. In some cases, said amino acid sequence is selected from the group consisting of SEQ ID NOs: 179, 185, 186, 187, 188, 191, 193, 194, and 195. In some cases, said at least one amino acid substitution to SEQ ID NO: 1 is selected from the group consisting of A53L, A53T, A53V, A53I, A53M, A53Q, A53K, and A53S. In some cases, said amino acid sequence is selected from the group consisting of SEQ ID NOs: 185, 186, 187, 188, 191, 193, 194, and 195.
BRIEF DESCRIPTION OF THE FIGURES FIG. 1 shows the total nMol of prenylated products produced by ORF2 triple mutants using OA as substrate and GPP as donor.
FIG. 2 shows % CBGA produced by ORF2 triple mutants using OA as substrate and GPP as donor.
FIG. 3 shows % enzymatic activity of ORF2 triple mutants using OA as substrate and GPP as donor
FIG. 4 shows CBGA production potential of ORF2 triple mutants using OA as substrate and GPP as donor.
FIG. 5 shows a cluster map of ORF2 triple mutants clustered based on CBGA production potential and %5-GOA produced, using OA as substrate and GPP as donor
FIG. 6 shows the analysis of ORF2 enzymatic function including total nMols of prenylated products produced (FIG. 6A) and % CBGA produced (FIG. 6B) by mutants derived from the breakdown of ORF2 triple mutant clone E09.
FIG. 7 shows the analysis of ORF2 enzymatic function including total nMols of prenylated products produced (FIG. 7A) and % CBGA produced (FIG. 7B) by mutants derived from the breakdown of ORF2 triple mutant clone H02.
FIG. 8 shows the analysis of ORF2 enzymatic function including total nMols of prenylated products produced (FIG. 8A) and % CBGA produced (FIG. 8B) by mutants derived from the breakdown of ORF2 triple mutant clone A04.
FIG. 9 shows the analysis of ORF2 enzymatic function including total nMols of prenylated products produced (FIG. 9A) and % CBGA produced (FIG. 9B) by mutants derived from the breakdown of ORF2 triple mutant clone D04.
FIG. 10 shows the analysis of ORF2 enzymatic function including total nMols of prenylated products produced (FIG. 10A) and % CBGA produced (FIG. 10B) by mutants derived from the breakdown of ORF2 triple mutant clone A09.
FIG. 11 shows the analysis of ORF2 enzymatic function including total nMols of prenylated products produced (FIG. 11A) and % CBGA produced (FIG. 11B) by mutants derived from the breakdown of ORF2 triple mutant clone C05.
FIG. 12 shows the analysis of ORF2 enzymatic function including total nMols of prenylated products produced (FIG. 12A) and % CBGA produced (FIG. 12B) by mutants derived from the breakdown of ORF2 triple mutant clone H011.
FIG. 13 shows the analysis of ORF2 enzymatic function including total nMols of prenylated products produced (FIG. 13A) and % CBGA produced (FIG. 13B) by mutants derived from the breakdown of ORF2 triple mutant clone F09.
FIG. 14 shows the analysis of ORF2 enzymatic function including total nMols of prenylated products produced (FIG. 14A) and % CBGA produced (FIG. 14B) by mutants derived from the breakdown of ORF2 triple mutant clone D011.
FIG. 15 shows the analysis of ORF2 enzymatic function including total nMols of prenylated products produced (FIG. 15A) and % CBGA produced (FIG. 15B) by mutants derived from the breakdown of ORF2 triple mutant clone G05.
FIG. 16 shows the analysis of enzymatic activity of site-saturated ORF2 mutants of Q295 using OA as substrate and GPP as donor, including total nMols of prenylated products produced (FIG. 16A); CBGA production potential (FIG. 16B); and 5-GOA production potential (FIG. 16C).
FIG. 17 shows the analysis of enzymatic activity of site-saturated ORF2 mutants of Q161 using OA as substrate and GPP as donor, including total nMols of prenylated products produced (FIG. 17A); CBGA production potential (FIG. 17B); and 5-GOA production potential (FIG. 17C).
FIG. 18 shows the analysis of enzymatic activity of site-saturated ORF2 mutants of 5214 using OA as substrate and GPP as donor, including total nMols of prenylated products produced (FIG. 18A); CBGA production potential (FIG. 18B); and 5-GOA production potential (FIG. 18C).
FIG. 19 shows the total nMol of prenylated products produced (using OA as substrate and GPP as donor) by ORF2 stacking mutants.
FIG. 20 shows the % enzymatic activity (using OA as substrate and GPP as donor) of ORF2 stacking mutants.
FIG. 21 shows the % CBGA produced (using OA as substrate and GPP as donor) by ORF2 stacking mutants.
FIG. 22 shows the CBGA production potential (using OA as substrate and GPP as donor) of ORF2 stacking mutants.
FIG. 23 shows the % ORF2 activity (using OA as substrate and GPP as donor) of A53 site saturation ORF2 mutants.
FIG. 24 shows % ORF2 activity (using OA as substrate and GPP as donor) of Q161 site saturation ORF2 mutants.
FIG. 25 shows the total nMol of prenylated products produced by ORF2 mutants using DVA as substrate and GPP as donor.
FIG. 26 shows % CBGVA and %5-GDVA produced by ORF2 mutants using DVA as substrate and GPP as donor.
FIG. 27 shows the % enzymatic activity of ORF2 mutants using DVA as substrate and GPP as donor.
FIG. 28 shows the CBGVA production potential of ORF2 mutants using DVA as substrate and GPP as donor.
FIG. 29 shows a cluster map of ORF2 mutants clustered based on CBGVA production potential and % GDVA produced, using DVA as substrate and GPP as donor
FIG. 30 shows the total amount of prenylated products (FIG. 30A) and % CBGVA in prenylated products (FIG. 30B) of A09 ORF2 mutant using DVA as substrate and GPP as donor.
FIG. 31 shows the total amount of prenylated products (FIG. 31A) and % CBGVA in prenylated products (FIG. 1B) of H02 ORF2 mutant using DVA as substrate and GPP as donor.
FIG. 32 shows the total amount of prenylated products (FIG. 32A) and % CBGVA in prenylated products (FIG. 32B) of C05 ORF2 mutant using DVA as substrate and GPP as donor.
FIG. 33 shows the total amount of prenylated products (FIG. 33A) and % CBGVA in prenylated products (FIG. 33B) of E09 ORF2 mutant using DVA as substrate and GPP as donor.
FIG. 34 shows the total amount of prenylated products (FIG. 34A) and % CBGVA in prenylated products (FIG. 34B) of D04 ORF2 mutant using DVA as substrate and GPP as donor.
FIG. 35 shows the total amount of prenylated products (FIG. 35A) and % CBGVA in prenylated products (FIG. 35B) of H11 ORF2 mutant using DVA as substrate and GPP as donor.
FIG. 36 shows the total amount of prenylated products (FIG. 36A) and % CBGVA in prenylated products (FIG. 36B) of A04 ORF2 mutant using DVA as substrate and GPP as donor.
FIG. 37 shows the total amount of prenylated products (FIG. 37A) and % CBGVA in prenylated products (FIG. 37B) of G05 ORF2 mutant using DVA as substrate and GPP as donor.
FIG. 38 shows the total amount of prenylated products (FIG. 38A) and % CBGVA in prenylated products (FIG. 38B) of F09 ORF2 mutant using DVA as substrate and GPP as donor.
FIG. 39 shows the total amount of prenylated products (FIG. 39A) and % CBGVA in prenylated products (FIG. 39B) of D11 ORF2 mutant using DVA as substrate and GPP as donor.
FIG. 40 shows the total amount of prenylated products (FIG. 40A); CBGVA production potential (FIG. 40B); and %5-GDVA production potential (FIG. 40C) of site saturated mutants of Q295.
FIG. 41 shows the total amount of prenylated products (FIG. 41A); CBGVA production potential (FIG. 41B); and %5-GDVA production potential (FIG. 41C) of site saturated mutants of Q161.
FIG. 42 shows the total amount of prenylated products (FIG. 42A); CBGVA production potential (FIG. 42B); and %5-GDVA production potential (FIG. 42C) of site saturated mutants of S214.
FIG. 43 shows total nMol of prenylated products produced (using DVA as substrate and GPP as donor) by ORF2 stacking mutants.
FIG. 44 shows the % enzymatic activity using DVA substrate and GPP as donor of ORF2 stacking mutants.
FIG. 45 shows the % CBGVA produced (using DVA as substrate and GPP as donor) by ORF2 stacking mutants
FIG. 46 shows that CBGVA production potential of the new ORF2 mutants generated by stacking the selected mutations.
FIG. 47 shows % enzymatic activity (using DVA as substrate and GPP as donor) of ORF2 mutants derived from breakdown of A09 mutant clone.
FIG. 48 shows % enzymatic activity (using DVA as substrate and GPP as donor) of ORF2 mutants derived from breakdown of E09 mutant clone.
FIG. 49 shows % enzymatic activity (using DVA as substrate and GPP as donor) of Q295 site-saturated ORF2 mutants.
FIG. 50 shows % enzymatic activity (using DVA as substrate and GPP as donor) of A53 site-saturated ORF2 mutants.
FIG. 51 shows % enzymatic activity (using DVA as substrate and GPP as donor) of Q161 site-saturated ORF2 mutants.
FIG. 52 shows % enzymatic activity (using DVA as substrate and GPP as donor) of stacking ORF2 mutants.
FIG. 53 shows the total nMol of prenylated products produced by ORF2 triple mutants using O as substrate and GPP as donor.
FIG. 54 shows % CBG and % GO produced by ORF2 triple mutants using O as substrate and GPP as donor.
FIG. 55 shows % enzymatic activity (using O as substrate and GPP as donor) of ORF2 triple mutants.
FIG. 56 shows the CBG production potential (using O as substrate and GPP as donor) of ORF2 triple mutants.
FIG. 57 shows a cluster graph of CBG production potential of ORF2 triple mutants vs. % GO produced by ORF2 triple mutants (using O as substrate and GPP as donor).
FIG. 58 shows the analysis of ORF2 enzymatic function (using O as substrate and GPP as donor)—including total nMol of prenylated products (FIG. 58A) and % CBG produced (FIG. 58B)—of ORF2 mutants derived from breakdown of E09 triple mutant.
FIG. 59 shows the analysis of ORF2 enzymatic function (using O as substrate and GPP as donor)—including total nMol of prenylated products (FIG. 59A) and % CBG produced (FIG. 59B)—of ORF2 mutants derived from breakdown of A09 triple mutant.
FIG. 60 shows the analysis of ORF2 enzymatic function (using O as substrate and GPP as donor)—including total nMol of prenylated products (FIG. 60A) and % CBG produced (FIG. 60B)—of ORF2 mutants derived from breakdown of A04 triple mutant.
FIG. 61 shows the analysis of ORF2 enzymatic function (using O as substrate and GPP as donor)—including total nMol of prenylated products (FIG. 61A) and % CBG produced (FIG. 61B)—of ORF2 mutants derived from breakdown of H011 triple mutant.
FIG. 62 shows the analysis of ORF2 enzymatic function (using O as substrate and GPP as donor)—including total nMol of prenylated products (FIG. 62A) and % CBG produced (FIG. 62B)—of ORF2 mutants derived from breakdown of H02 triple mutant.
FIG. 63 shows the analysis of ORF2 enzymatic function (using O as substrate and GPP as donor)—including total nMol of prenylated products (FIG. 63A) and % CBG produced (FIG. 63B)—of ORF2 mutants derived from breakdown of D04 triple mutant.
FIG. 64 shows the analysis of ORF2 enzymatic function (using O as substrate and GPP as donor)—including total nMol of prenylated products (FIG. 64A) and % CBG produced (FIG. 64B)—of ORF2 mutants derived from breakdown of C05 triple mutant.
FIG. 65 shows the analysis of ORF2 enzymatic function (using O as substrate and GPP as donor)—including total nMol of prenylated products (FIG. 65A) and % CBG produced (FIG. 65B)—of ORF2 mutants derived from breakdown of F09 triple mutant.
FIG. 66 shows the analysis of ORF2 enzymatic function (using O as substrate and GPP as donor)—including total nMol of prenylated products (FIG. 66A) and % CBG produced (FIG. 66B)—of ORF2 mutants derived from breakdown of D11 triple mutant.
FIG. 67 shows the analysis of ORF2 enzymatic function (using O as substrate and GPP as donor)—including total nMol of prenylated products (FIG. 67A) and % CBG produced (FIG. 67B)—of ORF2 mutants derived from breakdown of G05 triple mutant.
FIG. 68 shows the analysis of ORF2 enzymatic function (using O as substrate and GPP as donor)—including total nMol of prenylated products (FIG. 68A) and CBG production potential (FIG. 68B) —of ORF2 mutants carrying site saturation Q295 mutations.
FIG. 69 shows 5-GO production potential (using O as substrate and GPP as donor) of ORF2 mutants carrying site saturation Q295 mutations.
FIG. 70 shows the analysis of ORF2 enzymatic function (using O as substrate and GPP as donor)—including total nMol of prenylated products (FIG. 70A) and CBG production potential (FIG. 70B) —of ORF2 mutants carrying site saturation Q161 mutations.
FIG. 71 shows 5-GO production potential (using O as substrate and GPP as donor) of ORF2 mutants carrying site saturation Q161 mutations.
FIG. 72 shows the analysis of ORF2 enzymatic function (using O as substrate and GPP as donor)—including total nMol of prenylated products (FIG. 72A) and CBG production potential (FIG. 72B) —of ORF2 mutants carrying site saturation 5214 mutations.
FIG. 73 shows 5-GO production potential (using O as substrate and GPP as donor) of ORF2 mutants carrying site saturation 5214 mutations.
FIG. 74 shows the total nMol of prenylated products produced (using O as substrate and GPP as donor) by ORF2 stacking mutants.
FIG. 75 shows the % enzymatic activity (using O as substrate and GPP as donor) of ORF2 stacking mutants.
FIG. 76 shows the % CBG produced (using O as substrate and GPP as donor) by ORF2 stacking mutants.
FIG. 77 shows the CBG production potential (using O as substrate and GPP as donor) of ORF2 stacking mutants.
FIG. 78 shows % activity (using O as substrate and GPP as donor) of ORF2 mutants resulting from breakdown of tripe mutant clone E09.
FIG. 79 shows % activity (using O as substrate and GPP as donor) of ORF2 mutants resulting from breakdown of tripe mutant clone A09.
FIG. 80 shows % ORF2 activity (using O as substrate and GPP as donor) of Q161 site saturation ORF2 mutants.
FIG. 81: % ORF2 activity (using O as substrate and GPP as donor) of A53 site saturation ORF2 mutants.
FIG. 82 shows ORF-2 activity (using OA as substrate and GPP as donor) of A53-S214R ORF2 double mutants.
FIG. 83 shows ORF-2 activity (using OA as substrate and GPP as donor) of Q161S-Q295L ORF2 double mutants.
FIG. 84: ORF-2 activity (using OA as substrate and GPP as donor) of Q161S-Q295F ORF2 double mutants.
FIG. 85: ORF-2 activity (using OA as substrate and GPP as donor) of S177W-Q295A ORF2 double mutants.
FIG. 86: ORF-2 activity (using DVA as substrate and GPP as donor) of A53T-S214R ORF2 double mutants.
FIG. 87: ORF-2 activity (using DVA as substrate and GPP as donor) of Q161S-Q295L ORF2 double mutants.
FIG. 88: ORF-2 activity (using DVA as substrate and GPP as donor) of S177W-Q295A ORF2 double mutants.
FIG. 89: ORF-2 activity (using DVA as substrate and GPP as donor) of Q161S-Q295F ORF2 double mutants.
FIG. 90: ORF-2 activity (using DVA as substrate and GPP as donor) of A53T-Q295F ORF2 double mutants.
FIG. 91: ORF-2 activity (using DVA as substrate and GPP as donor) of Q161S-S177W ORF2 double mutants.
FIG. 92: ORF-2 activity (using O as substrate and GPP as donor) of Q161S-Q295L ORF2 double mutants.
FIG. 93: ORF-2 activity (using O as substrate and GPP as donor) of A53T-Q161S ORF2 double mutants.
FIG. 94: ORF-2 activity (using O as substrate and GPP as donor) of Q161S-Q295F ORF2 double mutants.
FIG. 95: ORF-2 activity (using O as substrate and GPP as donor) of A53T-Q295F ORF2 double mutants.
FIG. 96: ORF-2 activity (using O as substrate and GPP as donor) of A53T-V294A ORF2 double mutants.
FIG. 97. Analysis of enzymatic activity of site-saturated ORF2 mutants of Y288 using OA as substrate and GPP as donor—including total prenylated products (FIG. 97A); CBGA production potential (FIG. 97B); and 5GOA production potential (FIG. 97C).
FIG. 98. Analysis of enzymatic activity of site-saturated ORF2 mutants of Y288 using DVA as substrate and GPP as donor—including total prenylated products (FIG. 98A); CBGVA production potential (FIG. 98B); and 5-GDVA production potential (FIG. 98C).
FIG. 99. Analysis of enzymatic activity of site-saturated ORF2 mutants of Y288 using O as substrate and GPP as donor—including total prenylated products (FIG. 99A); CBG production potential (FIG. 99B); and 5-GO production potential (FIG. 99C).
FIG. 100: Analysis of ORF-2 enzymatic function of single mutants derived from the breakdown of ORF-2 triple mutant clone G12—including total prenylated products (FIG. 100A); and % CBGA produced (FIG. 100B).
FIG. 101: Analysis of ORF-2 enzymatic function of single mutants derived from the breakdown of ORF-2 triple mutant clone C06—including total prenylated products (FIG. 101A); and % CBGA produced (FIG. 101B).
FIG. 102: Analysis of ORF-2 enzymatic function of single mutants derived from the breakdown of ORF-2 triple mutant clone F08—including total prenylated products (FIG. 102A); and % CBGA produced (FIG. 102B).
FIG. 103: Analysis of ORF-2 enzymatic function of single mutants derived from the breakdown of ORF-2 triple mutant clone G12—including total prenylated products (FIG. 103A); and % CBGVA produced (FIG. 103B).
FIG. 104: Analysis of ORF-2 enzymatic function of single mutants derived from the breakdown of ORF-2 triple mutant clone C06—including total prenylated products (FIG. 104A); and % CBGVA produced (FIG. 104B).
FIG. 105: Analysis of ORF-2 enzymatic function of single mutants derived from the breakdown of ORF-2 triple mutant clone F08—including total prenylated products (FIG. 105A); and % CBGVA produced (FIG. 105B).
FIG. 106: Analysis of ORF-2 enzymatic function of single mutants derived from the breakdown of ORF-2 triple mutant clone G12—including total prenylated products (FIG. 106A); and % CBG produced (FIG. 106B).
FIG. 107: Analysis of ORF-2 enzymatic function of single mutants derived from the breakdown of ORF-2 triple mutant clone C06—including total prenylated products (FIG. 107A); and % CBG produced (FIG. 107B).
FIG. 108: Examples of benzoic acids and benzenediols applicable for prenylation by Orf2 mutants
FIG. 109: Structures of benzoic acids and benzenediols applicable for prenylation by Orf2 mutants
FIG. 110: Examples of other substrate molecules applicable for prenylation with Orf2 variants
FIG. 111: Total nMol of prenylated products produced by ORF2 triple mutants using OA as substrate and GPP as donor at pH 5.3
FIG. 112: % CBGA produced by ORF2 triple mutants using OA as substrate and GPP as donor pH 5.3
FIG. 113: % enzymatic activity of ORF2 triple mutants using OA as substrate and GPP as donor pH 5.3
FIG. 114: CBGA production potential of ORF2 triple mutants using OA as substrate and GPP as donor pH 5.3
FIG. 115: Cluster map of ORF2 triple mutants clustered based on CBGA production potential and %5-GOA produced, using OA as substrate and GPP as donor pH 5.3
FIG. 116: Analysis of ORF2 enzymatic activity (using OA as substrate and GPP as donor) for ORF2 mutants having either Q161H, S214H, or Q295H mutation.
DETAILED DESCRIPTION Definitions As used herein, and in the appended claims, the singular forms “a”, “an”, and “the” include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to “a protein” can refer to one protein or to mixtures of such protein, and reference to “the method” includes reference to equivalent steps and/or processes known to those skilled in the art, and so forth.
As used herein, the term “about” or “approximately” when preceding a numerical value indicates the value plus or minus a range of 10%. For example, “about 100” encompasses 90 and 110.
The term “wild type”, abbreviated as “WT”, is a term of the art understood by skilled persons and means the typical form of an organism, strain, gene, protein, or characteristic as it occurs in nature as distinguished from mutant or variant forms. For example, a WT protein is the typical form of that protein as it occurs in nature.
The term “mutant protein” is a term of the art understood by skilled persons and refers to a protein that is distinguished from the WT form of the protein on the basis of the presence of amino acid modifications, such as, for example, amino acid substitutions, insertions and/or deletions.
Amino acid modifications may be amino acid substitutions, amino acid deletions and/or amino acid insertions. Amino acid substitutions may be conservative amino acid substitutions or non-conservative amino acid substitutions. A conservative replacement (also called a conservative mutation, a conservative substitution or a conservative variation) is an amino acid replacement in a protein that changes a given amino acid to a different amino acid with similar biochemical properties (e.g. charge, hydrophobicity and size). As used herein, “conservative variations” refer to the replacement of an amino acid residue by another, biologically similar residue. Examples of conservative variations include the substitution of one hydrophobic residue such as isoleucine, valine, leucine or methionine for another; or the substitution of one polar residue for another, such as the substitution of arginine for lysine, glutamic for aspartic acids, or glutamine for asparagine, and the like. Other illustrative examples of conservative substitutions include the changes of: alanine to serine; arginine to lysine; asparagine to glutamine or histidine; aspartate to glutamate; cysteine to serine; glutamine to asparagine; glutamate to aspartate; glycine to praline; histidine to asparagine or glutamine; isoleucine to leucine or valine; leucine to valine or isoleucine; lysine to arginine, glutamine, or glutamate; methionine to leucine or isoleucine; phenylalanine to tyrosine, leucine or methionine; serine to threonine; threonine to serine; tryptophan to tyrosine; tyrosine to tryptophan or phenylalanine; valine to isoleucine or leucine, and the like.
Amino acid substitution, interchangeably referred to as amino acid replacement, at a specific position on the protein sequence is denoted herein in the following manner: “one letter code of the WT amino acid residue-amino acid position-one letter code of the amino acid residue that replaces this WT residue”. For example, an ORF2 polypeptide which is a Q295F mutant refers to an ORF2 polypeptide in which the wild type residue at the 295th amino acid position (Q or glutamine) is replaced with F or phenylalanine. Some mutants have more than one amino acid substitutions, for example, mutant L174V_S177E refers to an ORF2 polypeptide in which the wild type residue at the 174th amino acid position (L or leucine) is replaced with V or valine; and the wild type residue at the 177th amino acid position (S or serine) is replaced with E or glutamic acid.
The modified peptides can be chemically synthesized, or the isolated gene can be site-directed mutagenized, or a synthetic gene can be synthesized and expressed in bacteria, yeast, baculovirus, tissue culture, and the like.
As used herein, “total prenylated products” produced refers to the sum of nMols of the various prenylated products produced by an enzyme in a set period of time. For instance, when OA is used as a substrate and GPP is used as a donor, then the “total prenylated products” refers to a sum of the nMol of CBGA and the nMol of 5-GOA produced by the prenyltranferase enzyme ORF2 in a set period of time.
As used herein, “% prenylated product 1” within total prenylated products is calculated using the equation: nMol of prenylated product 1/[nMol of total prenylated products]. For example, “% CBGA” is calculated using the equation: nMol of CBGA/[nMol of CBGA+5-GOA]. Also, as an example, “%5-GOA” within prenylated products is calculated using the equation: nMol of 5-GOA/[nMol of CBGA+5-GOA].
As used herein, % enzymatic activity of an ORF2 mutant is calculated using the equation: total prenylated products produced by a mutant/total prenylated products produced by wild-type ORF2. For example, wild-type ORF2 has 100% enzyme activity.
As used herein, the production or production potential of a prenylated product 1 is calculated using the formula: % product 1 among total prenylated products * % enzymatic activity. For example, “CBGA production potential” (used interchangeably with “CBGA production”) is calculated using the equation: % CBGA among total prenylated products * % enzymatic activity. Also, as an example, “5-GOA production potential” (used interchangeably with “5-GOA production”) is calculated using the equation: %5-GOA among total prenylated products * enzymatic activity.
A “vector” is used to transfer genetic material into a target cell. Vectors include, but are not limited to, nucleic acid molecules that are single-stranded, double-stranded, or partially double-stranded; nucleic acid molecules that comprise one or more free ends, no free ends (e.g. circular); nucleic acid molecules that comprise DNA, RNA, or both; and other varieties of polynucleotides known in the art. One type of vector is a “plasmid,” which refers to a circular double stranded DNA loop into which additional DNA segments can be inserted, such as by standard molecular cloning techniques. Another type of vector is a viral vector, wherein virally-derived DNA or RNA sequences are present in the vector for packaging into a virus (e.g., retroviruses, adenoviruses, lentiviruses, and adeno-associated viruses). In embodiments, a viral vector may be replication incompetent. Viral vectors also include polynucleotides carried by a virus for transfection into a host cell. Certain vectors are capable of autonomous replication in a host cell into which they are introduced (e.g. bacterial vectors having a bacterial origin of replication and episomal mammalian vectors). Other vectors (e.g., non-episomal mammalian vectors) are integrated into the genome of a host cell upon introduction into the host cell, and thereby are replicated along with the host genome. Moreover, certain vectors are capable of directing the expression of genes to which they are operatively-linked. Such vectors are referred to herein as “expression vectors.” Common expression vectors of utility in recombinant DNA techniques are often in the form of plasmids.
As used herein “sequence identity” refers to the extent to which two optimally aligned polynucleotides or polypeptide sequences are invariant throughout a window of alignment of components, e.g. nucleotides or amino acids. An “identity fraction” for aligned segments of a test sequence and a reference sequence is the number of identical components which are shared by the two aligned sequences divided by the total number of components in the reference sequence segment, i.e. the entire reference sequence or a smaller defined part of the reference sequence. “Percent identity” is the identity fraction times 100. Comparison of sequences to determine percent identity can be accomplished by a number of well-known methods, including for example by using mathematical algorithms, such as, for example, those in the BLAST suite of sequence analysis programs.
Cannabinoid Synthesis The biosynthesis of cannabinoids often starts with the short-chain fatty acid, hexanoic acid. Initially, the fatty acid is converted to its coenzyme A (CoA) form by the activity of an acyl activating enzyme. Subsequently, olivetolic acid (OA) is biosynthesized by the action of a type III polyketide synthase (PKS), and, in some cases, a polyketide cyclase (olivetolic acid cyclase [OAC]).
A geranyl diphosphate:olivetolate geranyltransferase, named cannabigerolic acid synthase (CBGAS), is responsible for the C-alkylation by geranyl diphosphate (GPP) to CBGA. Subsequently, the monoterpene moiety of CBGA is often stereoselectively cyclized by three different enzymes cannabichromenic acid synthase (CBCAS), cannabidiolic acid synthase (CBDAS) and tetrahydrocannabinolic acid synthase (THCAS) to synthesize cannabichromenic acid (CBCA), cannabidiolic acid (CBDA) and Δ9-THCA, respectively.
The central precursor for cannabinoid biosynthesis, CBGA, is synthesized by the aromatic prenyltransferase CBGAS by the condensation of GPP and OA. In considering the biosynthesis of cannabinoids in a heterologous system, one major challenge is that CBGAS (e.g. CsPT1 and CsPT4) is an integral membrane protein, making high titer of functional expressed protein in E. coli and other heterologous systems unlikely. Besides the integral membrane prenyltransferases found in plants, soluble prenyltransferases are found in fungi and bacteria. For instance, Streptomyces sp. strain CL190 produces a soluble prenyltransferase NphB or ORF2, which is specific for GPP as a prenyl donor and exhibits broad substrate specificity towards aromatic substrates. When expressed in E. coli, ORF2 of SEQ ID NO:2 is as a 33 kDa soluble, monomeric protein having 307 residues. Further details about ORF2 and other aromatic prenyltransferases may be found in U.S. Pat. Nos. 7,361,483; 7,544,498; and 8,124,390, each of which is incorporated herein by reference in its entirety for all purposes.
ORF2 is a potential alternative to replace the native CBGAS in a biotechnological production of cannabinoids and other prenylated aromatic compounds. However, the wild type ORF2 enzyme produces a large amount of 5-geranyl olivetolate (5-GOA) and only a minor amount of CBGA, the latter of which is the desired product for cannabinoid biosynthesis.
This disclosure provides ORF2 mutants, engineered by the inventors to produce higher ratios of CBGA to 5-GOA (and other compounds), as compared to WT ORF2; as well as ORF2 mutants which have been engineered to catalyze reactions using a desired substrate and to produce higher amounts of a desired product.
ORF2 Mutants The disclosure provides recombinant polypeptides comprising an amino acid sequence with at least about 70% identity to the amino acid sequence of WT ORF2 (set forth in SEQ ID NO: 1). In some aspects, the polypeptides disclosed herein may have a sequence identity of about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 96%, about 97%, about 98%, about 99%, or about 99.5% identity to the amino acid sequence of WT ORF2 (set forth in SEQ ID NO: 1). In some aspects, the mutant ORF2 polypeptides disclosed herein may comprise a modification at one or more amino acids, as compared to the WT ORF2 sequence. In some aspects, the mutant ORF2 polypeptides disclosed herein may comprise a modification at 1 amino acid, 2 amino acids, 3 amino acids, 4 amino acids, 5 amino acids, 6 amino acids, 7 amino acids, 8 amino acids, 9 amino acids, 10 amino acids, 11 amino acids, 12 amino acids, 13 amino acids, 14 amino acids, 15 amino acids, 16 amino acids, 17 amino acids, 18 amino acids, 19 amino acids, 20 amino acids, 21 amino acids, 22 amino acids, 23 amino acids, 24 amino acids, 25 amino acids, 26 amino acids, 27 amino acids, 28 amino acids, 29 amino acids, 30 amino acids, 31 amino acids, 32 amino acids, 33 amino acids, 34 amino acids, 35 amino acids, or 36 amino acids, as compared to the WT ORF2 sequence.
In some aspects, mutant ORF2 polypeptides disclosed herein comprise a modification in one or more amino acid residues selected from the group consisting of the following amino acid residues, A17, C25, Q38, V49, A53, M106, A108, E112, K118, K119, Y121, F123, Q161, M162, D166, N173, L174, S177, G205, C209, F213, S214, Y216, L219, D227, R228, C230, A232, V271, L274, Y283, G286, Y288, V294, Q295, and L298 of the WT ORF2 polypeptide. For instance, the mutant ORF2 polypeptides disclosed herein may comprise an amino acid modification at 1 amino acid, 2 amino acids, 3 amino acids, 4 amino acids, 5 amino acids, 6 amino acids, 7 amino acids, 8 amino acids, 9 amino acids, 10 amino acids, 11 amino acids, 12 amino acids, 13 amino acids, 14 amino acids, 15 amino acids, 16 amino acids, 17 amino acids, 18 amino acids, 19 amino acids, 20 amino acids, 21 amino acids, 22 amino acids, 23 amino acids, 24 amino acids, 25 amino acids, 26 amino acids, 27 amino acids, 28 amino acids, 29 amino acids, 30 amino acids, 31 amino acids, 32 amino acids, 33 amino acids, 34 amino acids, 35 amino acids, or 36 amino acids selected from the group consisting of the following amino acid residues, A17, C25, Q38, V49, A53, M106, A108, E112, K118, K119, Y121, F123, Q161, M162, D166, N173, L174, S177, G205, C209, F213, S214, Y216, L219, D227, R228, C230, A232, V271, L274, Y283, G286, Y288, V294, Q295, and L298 of the WT ORF2 polypeptide.
In some aspects, the mutant ORF2 polypeptides disclosed herein may comprise an amino acid substitution of at least one amino acid residue selected from the group consisting of A17, C25, Q38, V49, A53, M106, A108, E112, K118, K119, Y121, F123, Q161, M162, D166, N173, L174, S177, G205, C209, F213, S214, Y216, L219, D227, R228, C230, A232, V271, L274, Y283, G286, Y288, V294, Q295, and L298. For instance, the mutant ORF2 polypeptides disclosed herein may comprise an amino acid substitution of 1 amino acid, 2 amino acids, 3 amino acids, 4 amino acids, 5 amino acids, 6 amino acids, 7 amino acids, 8 amino acids, 9 amino acids, 10 amino acids, 11 amino acids, 12 amino acids, 13 amino acids, 14 amino acids, 15 amino acids, 16 amino acids, 17 amino acids, 18 amino acids, 19 amino acids, 20 amino acids, 21 amino acids, 22 amino acids, 23 amino acids, 24 amino acids, 25 amino acids, 26 amino acids, 27 amino acids, 28 amino acids, 29 amino acids, 30 amino acids, 31 amino acids, 32 amino acids, 33 amino acids, 34 amino acids, 35 amino acids, or 36 amino acids selected from the group consisting of A17, C25, Q38, V49, A53, M106, A108, E112, K118, K119, Y121, F123, Q161, M162, D166, N173, L174, S177, G205, C209, F213, S214, Y216, L219, D227, R228, C230, A232, V271, L274, Y283, G286, Y288, V294, Q295, and L298.
In some aspects, the mutant ORF2 polypeptides disclosed herein comprise an amino acid sequence comprising at least one amino acid substitution, as compared to the amino acid sequence of WT ORF2, wherein the at least one amino acid substitution does not comprise an alanine substitution on an amino acid residue selected from the group consisting of 47, 64, 110, 121, 123, 126, 161, 175, 177, 214, 216, 288, 294 and 295.
In some aspects, the mutant ORF2 polypeptides disclosed herein comprise an amino acid sequence comprising at least one amino acid substitution, as compared to the amino acid sequence of WT ORF2, wherein at least one amino acid substitution is at a position selected from the group consisting of 1-46, 48-63, 65-109, 111-120, 122, 124, 125, 127-160, 162-174, 176, 178-213, 215, 217-287, 289-293, 296-307, on WT-ORF2.
In some aspects, the mutant ORF2 polypeptides disclosed herein comprise an amino acid sequence with at least about 70% identity (for instance, about 75%, about 80%, about 85%, about 90%, about 95%, about 96%, about 97%, about 98%, about 99%, or about 99.5% identity, inclusive of all values and subranges therebetween) to the amino acid sequence of SEQ ID Nos 2-300. In some aspects, the mutant ORF2 polypeptides disclosed herein comprise the amino acid sequence of SEQ ID Nos 2-300. In some aspects, the mutant ORF2 polypeptides disclosed herein consist of the amino acid sequence of SEQ ID Nos 2-300.
In some aspects, the mutant ORF2 polypeptides disclosed herein catalyze a reaction using GPP as a donor of the prenyl group. In some aspects, the mutant ORF2 polypeptides disclosed herein catalyze a reaction using any known substrate of ORF2, or a homologue thereof. In some aspects, the substrate is selected from the group consisting of olivetolic acid (OA), divarinolic acid (DVA) and olivetol (O). In some aspects, the substrate is any natural or synthetic phenolic acids with a 1,3-dihydroxyl motif, alternatively a resorcinol ring including but not limited to resveratrol, piceattanol and related stilbenes, naringenin, apigenin and related flavanones and flavones, respectively, Isoliquiritigenin, 2′-O-methylisoliquiritigenin and related chalcones, catechins and epi-catechins of all possible stereoisomers, biphenyl compounds such as 3,5-dihydroxy-biphenyl, benzophenones such as phlorobenzophenone, isoflavones such as biochanin A, genistein, and daidzein. For instance, the substrate may be any substrate listed in Tables A and B; and FIGS. 117-119.
TABLE A
Examples of ORF2 substrates which are benzoic acids and benzenediols
Tail Chain
IUPAC Chemical Name Common Name Length CAS#
2,4-dihydroxybenzoic acid β-Resorcylic acid 0-carbon 89-86-1
1,3-benzenediol resorcinol 0-carbon 108-46-3
2,4-dihydroxy-6-methylbenzoic o-orsellinic Acid 1-carbon 480-64-8
acid
1,3-Dihydroxy-5-methylbenzene Orcinol 1-carbon 504-15-4
2,4-Dihydroxy-6-aethyl- 2-carbon 4299-73-4
benzoesaeure
5-ethylbenzene-1,3-diol 2-carbon 4299-72-3
2,4-dihydroxy-6-propylbenzoic Divarinic Acid 3-carbon 4707-50-0
acid
5-propylbenzene-1,3-diol Divarin 3-carbon 500-49-2
2-butyl-4,6-dihydroxybenzoic 4-carbon 173324-41-9
acid
5-butylbenzene-1,3-diol 4-carbon 46113-76-2
2,4-dihydroxy-6-pentyl-benzoic Olivetolic Acid 5-carbon 491-72-5
acid;
5-pentylbenzene-1,3-diol Olivetol 5-carbon 500-66-3
5-hexylbenzene-1,3-diol 6-carbon 5465-20-3
2-hepty1-4,6-dihydroxy-benzoic sphaerophorolcarboxylic 7-carbon 6121-76-2
acid acid
5-heptylbenzene-1,3-diol Sphaerophorol 7-carbon 500-67-4
5-Dodecylbenzene-1,3-diol 12-carbon 72707-60-9
5-nonadecylbenzene-1,3-diol 19-carbon 35176-46-6
TABLE B
Examples of other aromatic compounds which are ORF2 substrates
IUPAC Chemical Name Common Name CAS#
1,3-Benzenediol resorcinol 108-46-3
3,4′,5-Trihydroxystilbene resveratrol 89-86-1
4′5-Tetrahydroxystilbene Piceatannol 4339-71-3
1,2-Diphenylethylene stilbene 103-30-0
2-Phenylbenzopyran-4-one flavone 525-82-6
2-Phenylchroman-4-one flavanone 487-26-3
1,3-benzenediol naringenin 108-46-3
5,7,4′-Trihydroxyflavone apigenin 8002-66-2
(E)-1-(2,4- Isoliquiritigenin 961-29-5
dihydroxyphenyl)-3-(4-
hydroxyphenyl)prop-2-en-1-
one
4,4′-dihydroxy-2′- 2′-O-Methylisoliquiritigenin 112408-67-0
methoxychalcone
1,3-Diphenylpropenone chalcone 94-41-7
(2R,3S)-2-(3,4- catechin 7295-85-4
Dihydroxyphenyl)chroman-
3,5,7-triol
(2R,3R)-2-(3,4- epi-catechin 7295-85-4
Dihydroxyphenyl)-3,5,7-
chromanetriol
Phenylbenzene biphenyl 92-52-4
5-Phenylresorcinol 3,5-Dihydroxybiphenyl 7028-41-3
diphenylmethanone benzophenone 119-61-9
3-phenyl-4H-chromen-4-one isoflavone 574-12-9
5,7-Dihydroxy-3-(4- biochanin A 491-80-5
methoxyphenyl)-4H-
chromen-4-one
4′,5,7-Trihydroxyisoflavone Genistein 690224-00-1
4′,7-Dihydroxyisoflavone Diadzein 486-66-8
4-Hydroxy-6-methyl-2H- Triacetic acid lactone 675-10-5
pyran-2-one
1,6-DHN 575-44-0
In some aspects, the products of ORF2 prenylation may further serve as substrates for ORF2. Therefore, the substrate may also be any product of an ORF2 prenylation reaction.
In some aspects, the mutant ORF2 polypeptides disclosed herein produce a higher amount of total nMol of prenylated products than WT ORF2. In some aspects, the mutant ORF2 polypeptides disclosed herein produce an amount of total nMol of prenylated products that is about 1% to about 1000% (for example, about 1%, about 5%, about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 100%, about 150%, about 200%, about 300%, about 400%, about 500%, about 600%, about 700%, about 800%, or about 900%), inclusive all the values and subranges that lie therebetween, higher than the amount of total nMol of prenylated products produced by WT ORF2.
In some aspects, the mutant ORF2 polypeptides disclosed herein have an enzymatic activity higher than WT ORF2. In some aspects, the mutant ORF2 polypeptides disclosed herein have an activity that is about 1% to about 1000% (for example, about 1%, about 5%, about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 100%, about 150%, about 200%, about 300%, about 400%, about 500%, about 600%, about 700%, about 800%, or about 900%), inclusive all the values and subranges that lie therebetween, higher than the enzymatic activity of WT ORF2.
ORF2 Mutants Catalyzing a Reaction Using OA as Substrate In some aspects the mutant ORF2 polypeptides disclosed herein catalyze the reaction converting OA and GPP to CBGA and 5-GOA.
In some aspects, the mutant ORF2 polypeptides disclosed herein produce a higher amount of total nMol of prenylated products than WT ORF2, wherein the mutant ORF2 polypeptides use OA as substrate and GPP as donor; and the prenylated products are CBGA and 5-GOA. In some aspects, the mutant ORF2 polypeptides disclosed herein produce an amount of total nMol of prenylated products that is about 1% to about 1000% (for example, about 1%, about 5%, about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 100%, about 150%, about 200%, about 300%, about 400%, about 500%, about 600%, about 700%, about 800%, or about 900%), inclusive all the values and subranges that lie therebetween, higher than the amount of total nMol of prenylated products produced by WT ORF2, wherein the prenylated products are CBGA and 5-GOA.
The ratio of CBGA to 5-GOA produced by WT ORF2 is about 20:80. In some aspects, the mutant ORF2 polypeptides disclosed herein produce a ratio of CBGA to 5-GOA that is higher than the ratio of CBGA to 5-GOA produced by WT ORF2 under the same conditions. In some aspects, the mutant ORF2 polypeptides disclosed herein produce a ratio of CBGA to 5-GOA that is lower than the ratio of CBGA to 5-GOA produced by WT ORF2 under the same conditions. In some embodiments, the ratio of CBGA to 5-GOA produced by the mutant ORF2 proteins of this disclosure may range from about 1:99 to up to 20:80; or more than about 20:80 to 99:1, for example about 5:95, about 10:90, about 15:85, about 25:75, about 30:70, about 35:65, about 40:60, about 45:55, about 50:50, about 55:45, about 60:40, about 65:35, about 70:30, about 75:25, about 80:20, about 85:15, about 90:10, about 95:5, or about 99:1, including all values and subranges that lie therebetween.
In some aspects, the mutant ORF2 polypeptides disclosed herein have an enzymatic activity higher than WT ORF2, wherein the mutant ORF2 polypeptides use OA as substrate and GPP as donor. In some aspects, the mutant ORF2 polypeptides disclosed herein have an activity that is about 1% to about 1000% (for example, about 1%, about 5%, about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 100%, about 150%, about 200%, about 300%, about 400%, about 500%, about 600%, about 700%, about 800%, or about 900%), inclusive all the values and subranges that lie therebetween, higher than the enzymatic activity of WT ORF2, wherein the mutant ORF2 polypeptides use OA as substrate and GPP as donor.
In some aspects, the mutant ORF2 polypeptides disclosed herein produce a higher % of CBGA than the WT ORF2. In some aspects, the mutant ORF2 polypeptides disclosed herein produce about 1% to about 1000% (for example, about 1%, about 5%, about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 100%, about 150%, about 200%, about 300%, about 400%, about 500%, about 600%, about 700%, about 800%, or about 900%), inclusive all the values and subranges that lie therebetween, higher % of CBGA than WT ORF2.
In some aspects, the mutant ORF2 polypeptides disclosed herein produce a lower % of CBGA than the WT ORF2. In some aspects, the mutant ORF2 polypeptides disclosed herein produce about 1% to about 100% (for example, about 1%, about 5%, about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, or about 100%), inclusive all the values and subranges that lie therebetween, lower % of CBGA than WT ORF2.
In some aspects, the mutant ORF2 polypeptides disclosed herein produce a higher % of 5-GOA than WT ORF2. In some aspects, the mutant ORF2 polypeptides disclosed herein produce about 1% to about 1000% (for example, about 1%, about 5%, about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 100%, about 150%, about 200%, about 300%, about 400%, about 500%, about 600%, about 700%, about 800%, or about 900%), inclusive all the values and subranges that lie therebetween, higher % of 5-GOA than the WT ORF2.
In some aspects, the mutant ORF2 polypeptides disclosed herein produce a lower % of 5-GOA than WT ORF2. In some aspects, the mutant ORF2 polypeptides disclosed herein produce about 1% to about 100% (for example, about 1%, about 5%, about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 100%), inclusive all the values and subranges that lie therebetween, lower % of 5-GOA than the WT ORF2.
In some aspects, the mutant ORF2 polypeptides disclosed herein have a CBGA production potential that is higher than that of WT ORF2. As used herein, CBGA production potential of an enzyme is calculated by multiplying the % CBGA in total prenylated products by the % activity calculated when the mutant ORF-2 uses OA as substrate and GPP as donor. In some aspects, the mutant ORF2 polypeptides disclosed herein have a CBGA production potential that is 1% to about 1000% (for example, about 1%, about 5%, about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 100%, about 150%, about 200%, about 300%, about 400%, about 500%, about 600%, about 700%, about 800%, or about 900%), inclusive all the values and subranges that lie therebetween, higher than that of WT ORF2.
In some aspects, the mutant ORF2 polypeptides disclosed herein produce a lower % of 5-GOA than WT ORF2; and have a CBGA production potential that is higher than that of WT ORF2.
In some aspects, the mutant ORF2 polypeptides disclosed herein have a 5-GOA production potential that is higher than that of WT ORF2. As used herein, 5-GOA production potential of an enzyme is calculated by multiplying the %5-GOA in total prenylated products by the % activity calculated when the mutant ORF-2 uses OA as substrate and GPP as donor. In some aspects, the mutant ORF2 polypeptides disclosed herein has a 5-GOA production potential that is 1% to about 1000% (for example, about 1%, about 5%, about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 100%, about 150%, about 200%, about 300%, about 400%, about 500%, about 600%, about 700%, about 800%, or about 900%), inclusive all the values and subranges that lie therebetween, higher than that of WT ORF2.
ORF2 Mutants Catalyzing a Reaction Using DVA as Substrate In some aspects, the mutant ORF2 polypeptides disclosed herein catalyze the reaction of converting DVA and GPP to CBGVA and 5-GDVA.
In some aspects, the mutant ORF2 polypeptides disclosed herein produce a higher amount of total nMol of prenylated products than WT ORF2, wherein the mutant ORF2 polypeptides use DVA as substrate and GPP as donor; and the prenylated products are CBGVA and 5-GDVA. In some aspects, the mutant ORF2 polypeptides disclosed herein produce an amount of total nMol of prenylated products that is about 1% to about 1000% (for example, about 1%, about 5%, about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 100%, about 150%, about 200%, about 300%, about 400%, about 500%, about 600%, about 700%, about 800%, or about 900%), inclusive all the values and subranges that lie therebetween, higher than the amount of total nMol of prenylated products produced by WT ORF2, wherein the prenylated products are CBGVA and 5-GDVA.
The ratio of CBGVA to 5-GDVA produced by WT ORF2 is about 16:84. In some aspects, the mutant ORF2 polypeptides disclosed herein produce a ratio of CBGVA to 5-GDVA that is higher than the ratio of CBGVA to 5-GDVA produced by WT ORF2 under the same conditions. In some aspects, the mutant ORF2 polypeptides disclosed herein produce a ratio of CBGVA to 5-GDVA that is lower than the ratio of CBGVA to 5-GDVA produced by WT ORF2 under the same conditions. In some embodiments, the ratio of CBGVA to 5-GDVA produced by the mutant ORF2 proteins of this disclosure may range from about 1:99 to up to about 16:84; or more than about 16:84 to about 99:1, for example about 5:95, about 10:90, about 15:85, about 20:80, about 25:75, about 30:70, about 35:65, about 40:60, about 45:55, about 50:50, about 55:45, about 60:40, about 65:35, about 70:30, about 75:25, about 80:20, about 85:15, about 90:10, about 95:5, or about 99:1, including all values and subranges that lie therebetween.
In some aspects, the mutant ORF2 polypeptides disclosed herein have an enzymatic activity higher than WT ORF2, wherein the mutant ORF2 polypeptides use DVA as substrate and GPP as donor. In some aspects, the mutant ORF2 polypeptides disclosed herein have an activity that is about 1% to about 1000% (for example, about 1%, about 5%, about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 100%, about 150%, about 200%, about 300%, about 400%, about 500%, about 600%, about 700%, about 800%, or about 900%), inclusive all the values and subranges that lie therebetween, higher than the enzymatic activity of WT ORF2, wherein the mutant ORF2 polypeptides use DVA as substrate and GPP as donor.
In some aspects, the mutant ORF2 polypeptides disclosed herein produce a higher % of CBGVA than the WT ORF2. In some aspects, the mutant ORF2 polypeptides disclosed herein produce about 1% to about 1000% (for example, about 1%, about 5%, about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 100%, about 150%, about 200%, about 300%, about 400%, about 500%, about 600%, about 700%, about 800%, or about 900%), inclusive all the values and subranges that lie therebetween, higher % of CBGVA than WT ORF2.
In some aspects, the mutant ORF2 polypeptides disclosed herein produce a lower % of CBGVA than the WT ORF2. In some aspects, the mutant ORF2 polypeptides disclosed herein produce about 1% to about 100% (for example, about 1%, about 5%, about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, or about 100%), inclusive all the values and subranges that lie therebetween, lower % of CBGVA than WT ORF2.
In some aspects, the mutant ORF2 polypeptides disclosed herein produce a higher % of 5-GDVA than WT ORF2. In some aspects, the mutant ORF2 polypeptides disclosed herein produce about 1% to about 1000% (for example, about 1%, about 5%, about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 100%, about 150%, about 200%, about 300%, about 400%, about 500%, about 600%, about 700%, about 800%, or about 900%), inclusive all the values and subranges that lie therebetween, higher % of 5-GDVA than the WT ORF2.
In some aspects, the mutant ORF2 polypeptides disclosed herein produce a lower % of 5-GDVA than WT ORF2. In some aspects, the mutant ORF2 polypeptides disclosed herein produce about 1% to about 100% (for example, about 1%, about 5%, about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 100%), inclusive all the values and subranges that lie therebetween, lower % of 5-GDVA than the WT ORF2.
In some aspects, the mutant ORF2 polypeptides disclosed herein have a CBGVA production potential is higher than that of WT ORF2. As used herein, CBGVA production potential of an enzyme is calculated by multiplying the % CBGVA in total prenylated products by the % activity calculated when the mutant ORF-2 uses DVA as substrate and GPP as donor. In some aspects, the mutant ORF2 polypeptides disclosed herein have a CBGVA production potential that is 1% to about 1000% (for example, about 1%, about 5%, about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 100%, about 150%, about 200%, about 300%, about 400%, about 500%, about 600%, about 700%, about 800%, or about 900%), inclusive all the values and subranges that lie therebetween, higher than that of WT ORF2.
In some aspects, the mutant ORF2 polypeptides disclosed herein produce a lower % of 5-GDVA than WT ORF2; and have a CBGVA production potential that is higher than that of WT ORF2.
In some aspects, the mutant ORF2 polypeptides disclosed herein have a 5-GDVA production potential that is higher than that of WT ORF2. As used herein, 5-GDVA production potential of an enzyme is calculated by multiplying the %5-GDVA in total prenylated products by the % activity calculated when the mutant ORF-2 uses DVA as substrate and GPP as donor. In some aspects, the mutant ORF2 polypeptides disclosed herein has a 5-GDVA production potential that is 1% to about 1000% (for example, about 1%, about 5%, about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 100%, about 150%, about 200%, about 300%, about 400%, about 500%, about 600%, about 700%, about 800%, or about 900%), inclusive all the values and subranges that lie therebetween, higher than that of WT ORF2.
ORF2 Mutants Catalyzing a Reaction Using O as Substrate In some aspects, the mutant ORF2 polypeptides disclosed herein catalyze the reaction of converting O and GPP to CBG and 5-GO. See FIG. 118, showing that ORF2 substrates include benzoic acids and benzenediols.
In some aspects, the mutant ORF2 polypeptides disclosed herein produce a higher amount of total nMol of prenylated products than WT ORF2, wherein the mutant ORF2 polypeptides use O as substrate and GPP as donor; and the prenylated products are CBG and 5-GO. In some aspects, the mutant ORF2 polypeptides disclosed herein produce an amount of total nMol of prenylated products that is about 1% to about 1000% (for example, about 1%, about 5%, about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 100%, about 150%, about 200%, about 300%, about 400%, about 500%, about 600%, about 700%, about 800%, or about 900%), inclusive all the values and subranges that lie therebetween, higher than the amount of total nMol of prenylated products produced by WT ORF2, wherein the prenylated products are CBG and 5-GO.
The ratio of CBG to 5-GO produced by WT ORF2 is about 62:38. In some aspects, the mutant ORF2 polypeptides disclosed herein produce a ratio of CBG to 5-GO that is higher than the ratio of CBG to 5-GO produced by WT ORF2 under the same conditions. In some aspects, the mutant ORF2 polypeptides disclosed herein produce a ratio of CBG to 5-GO that is lower than the ratio of CBG to 5-GO produced by WT ORF2 under the same conditions. In some embodiments, the ratio of CBG to 5-GO produced by the mutant ORF2 proteins of this disclosure may range from about 99:1 to up to about 62:38; or less than about 62:38 to about 1:99, for example about 5:95, about 10:90, about 15:85, about 20:80, about 25:75, about 30:70, about 35:65, about 40:60, about 45:55, about 50:50, about 55:45, about 60:40, about 65:35, about 70:30, about 75:25, about 80:20, about 85:15, about 90:10, about 95:5, or about 99:1, including all values and subranges that lie therebetween.
In some aspects, the mutant ORF2 polypeptides disclosed herein have an enzymatic activity higher than WT ORF2, wherein the mutant ORF2 polypeptides use O as substrate and GPP as donor. In some aspects, the mutant ORF2 polypeptides disclosed herein have an activity that is about 1% to about 1000% (for example, about 1%, about 5%, about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 100%, about 150%, about 200%, about 300%, about 400%, about 500%, about 600%, about 700%, about 800%, or about 900%), inclusive all the values and subranges that lie therebetween, higher than the enzymatic activity of WT ORF2, wherein the mutant ORF2 polypeptides use O as substrate and GPP as donor.
In some aspects, the mutant ORF2 polypeptides disclosed herein produce a higher % of CBG than the WT ORF2. In some aspects, the mutant ORF2 polypeptides disclosed herein produce about 1% to about 1000% (for example, about 1%, about 5%, about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 100%, about 150%, about 200%, about 300%, about 400%, about 500%, about 600%, about 700%, about 800%, or about 900%), inclusive all the values and subranges that lie therebetween, higher % of CBG than WT ORF2.
In some aspects, the mutant ORF2 polypeptides disclosed herein produce a lower % of CBG than the WT ORF2. In some aspects, the mutant ORF2 polypeptides disclosed herein produce about 1% to about 100% (for example, about 1%, about 5%, about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, or about 100%), inclusive all the values and subranges that lie therebetween, lower % of CBG than WT ORF2.
In some aspects, the mutant ORF2 polypeptides disclosed herein produce a higher % of 5-GO than WT ORF2. In some aspects, the mutant ORF2 polypeptides disclosed herein produce about 1% to about 1000% (for example, about 1%, about 5%, about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 100%, about 150%, about 200%, about 300%, about 400%, about 500%, about 600%, about 700%, about 800%, or about 900%), inclusive all the values and subranges that lie therebetween, higher % of 5-GO than the WT ORF2.
In some aspects, the mutant ORF2 polypeptides disclosed herein produce a lower % of 5-GO than WT ORF2. In some aspects, the mutant ORF2 polypeptides disclosed herein produce about 1% to about 100% (for example, about 1%, about 5%, about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 100%), inclusive all the values and subranges that lie therebetween, lower % of 5-GO than the WT ORF2.
In some aspects, the mutant ORF2 polypeptides disclosed herein have a CBG production potential is higher than that of WT ORF2. As used herein, CBG production potential of an enzyme is calculated by multiplying the % CBG in total prenylated products by the % activity calculated when the mutant ORF-2 uses O as substrate and GPP as donor. In some aspects, the mutant ORF2 polypeptides disclosed herein have a CBG production potential that is 1% to about 1000% (for example, about 1%, about 5%, about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 100%, about 150%, about 200%, about 300%, about 400%, about 500%, about 600%, about 700%, about 800%, or about 900%), inclusive all the values and subranges that lie therebetween, higher than that of WT ORF2.
In some aspects, the mutant ORF2 polypeptides disclosed herein produce a lower % of 5-GO than WT ORF2; and have a CBG production potential that is higher than that of WT ORF2.
In some aspects, the mutant ORF2 polypeptides disclosed herein have a 5-GO production potential is higher than that of WT ORF2. As used herein, 5-GO production potential of an enzyme is calculated by multiplying the %5-GO in total prenylated products by the % activity calculated when the mutant ORF-2 uses O as substrate and GPP as donor. In some aspects, the mutant ORF2 polypeptides disclosed herein has a 5-GO production potential that is 1% to about 1000% (for example, about 1%, about 5%, about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 100%, about 150%, about 200%, about 300%, about 400%, about 500%, about 600%, about 700%, about 800%, or about 900%), inclusive all the values and subranges that lie therebetween, higher than that of WT ORF2.
ORF2 Mutants Catalyzing a Reaction at pH 5.3 In some aspects, the ORF2 mutants disclosed herein have improved function at a pH range of about 3 to about 8, for instance, about 3.5, about 4, about 4.5 about 5, about 5.5, about 6, about 6.5, about 7, about 7.5, about 8, including all values and subranges therebetween. In some aspects, the ORF2 mutants disclosed herein which have improved function at acidic pH may be used in combination with at least one other enzyme which has optimal function at acidic pH. In some aspects, the at least one enzyme is CBD synthase.
In some aspects, the mutant ORF2 polypeptides disclosed herein have improved enzymatic function (increased amount of prenylated products, increased % CBGA or 5-GOA produced, increased % activity and/or increased CBGA or 5-GOA production potential) at pH 5.3 compared to WT ORF2 at the same pH. WT ORF2 has little to no activity at this acidic pH. Therefore, in some aspects, the ORF2 mutants disclosed herein which have improved function at pH 5.3 may be used in situations where a low pH is needed to be maintained. For instance, CBD synthase has optimal pH at around 5.3. Therefore, the ORF2 mutants disclosed herein which have improved function at pH 5.3 may be used for carrying out single-pot reactions along with CBD synthase.
Polynucleotides, Vectors and Methods The disclosure provides isolated or purified polynucleotides that encode any one of the ORF2 mutant polypeptides disclosed herein. The disclosure provides polynucleotides comprising a nucleic acid sequence with at least about 80% identity (for instance, about 85%, about 90%, about 95%, about 96%, about 97%, about 98%, or about 99%, and inclusive of all values and subranges therebetween) to the nucleic acid sequence set forth in [[all the DNA SEQ ID Nos.]].
The disclosure provides a vector comprising any one of the ORF2 mutant polynucleotide sequences disclosed herein.
The disclosure further provides a host cell comprising any one of the vectors disclosed herein; any one of the polynucleotides disclosed herein; or any one of the polynucleotides encoding the ORF2 mutants disclosed herein. Non-limiting examples of host cells include microbial host cells, such as, for example, bacteria, E. coli, yeast, microalgae; non-microbial hosts, such as, for example, insect cells, mammalian cell culture, plant cultures; and whole terrestrial plants. In some aspects, expression of any one of the vectors disclosed herein; any one of the polynucleotides disclosed herein; or any one of the polynucleotides encoding the ORF2 mutants disclosed herein may be done ex vivo or in vitro. In some aspects, expression of any one of the vectors disclosed herein; any one of the polynucleotides disclosed herein; or any one of the polynucleotides encoding the ORF2 mutants disclosed herein may be done in cell-free systems.
The disclosure provides methods of producing any one of the mutant ORF2 polypeptides disclosed herein, comprising culturing the host cell comprising any one of the vectors disclosed herein, in a medium permitting expression of the mutant ORF2 polypeptide, and isolating or purifying the mutant ORF2 polypeptide from the host cell.
The disclosure provides methods of producing CBGA in a host cell, comprising expressing a mutant ORF2 polypeptide, wherein the mutant ORF2 polypeptide has a CBGA production potential that is higher than that of WT ORF2, and wherein the mutant ORF2 polypeptide produces a lower % of 5-GOA than WT ORF2. In some aspects, the CBGA is produced ex vivo or in vitro. In some aspects, CBGA is produced in cell-free systems. In some aspects, the CBGA is produced in host cells, such as, for example, microbial host cells, such as, for example, bacteria, E. coli, yeast, microalgae; non-microbial hosts, such as, for example, insect cells, mammalian cells, plant cells; and whole terrestrial plants. In some aspects, the CBGA is produced in the medium of host cell culture.
The disclosure provides methods of producing 5-GOA in a host cell, comprising expressing a mutant ORF2 polypeptide, wherein the mutant ORF2 polypeptide has a 5-GOA production potential that is higher than that of WT ORF2, and wherein the mutant ORF2 polypeptide produces a lower % of CBGA than WT ORF2. In some aspects, the 5-GOA is produced ex vivo or in vitro. In some aspects, 5-GOA is produced in cell-free systems. In some aspects, the 5-GOA is produced in host cells, such as, for example, microbial host cells, such as, for example, bacteria, E. coli, yeast, microalgae; non-microbial hosts, such as, for example, insect cells, mammalian cells, plant cells; and whole terrestrial plants. In some aspects, the 5-GOA is produced in the medium of host cell culture.
The disclosure provides methods of producing CBGVA in a host cell, comprising expressing a mutant ORF2 polypeptide, wherein the mutant ORF2 polypeptide has a CBGVA production potential that is higher than that of WT ORF2, and wherein the mutant ORF2 polypeptide produces a lower % of 5-GDVA than WT ORF2. In some aspects, the CBGVA is produced ex vivo or in vitro. In some aspects, CBGVA is produced in cell-free systems. In some aspects, the CBGVA is produced in host cells, such as, for example, microbial host cells, such as, for example, bacteria, E. coli, yeast, microalgae; non-microbial hosts, such as, for example, insect cells, mammalian cells, plant cells; and whole terrestrial plants. In some aspects, the CBGVA is produced in the medium of host cell culture.
The disclosure provides methods of producing 5-GDVA in a host cell, comprising expressing a mutant ORF2 polypeptide, wherein the mutant ORF2 polypeptide has a 5-GDVA production potential that is higher than that of WT ORF2, and wherein the mutant ORF2 polypeptide produces a lower % of CBGVA than WT ORF2. In some aspects, the 5-GDVA is produced ex vivo or in vitro. In some aspects, 5-GDVA is produced in cell-free systems. In some aspects, the 5-GDVA is produced in host cells, such as, for example, microbial host cells, such as, for example, bacteria, E. coli, yeast, microalgae; non-microbial hosts, such as, for example, insect cells, mammalian cells, plant cells; and whole terrestrial plants. In some aspects, the 5-GDVA is produced in the medium of host cell culture.
The disclosure provides methods of producing CBG in a host cell, comprising expressing a mutant ORF2 polypeptide, wherein the mutant ORF2 polypeptide has a CBG production potential that is higher than that of WT ORF2, and wherein the mutant ORF2 polypeptide produces a lower % of 5-GO than WT ORF2. In some aspects, the CBG is produced ex vivo or in vitro. In some aspects, CBG is produced in cell-free systems. In some aspects, the CBG is produced in host cells, such as, for example, microbial host cells, such as, for example, bacteria, E. coli, yeast, microalgae; non-microbial hosts, such as, for example, insect cells, mammalian cells, plant cells; and whole terrestrial plants. In some aspects, the CBG is produced in the medium of host cell culture.
The disclosure provides methods of producing 5-GO in a host cell, comprising expressing a mutant ORF2 polypeptide, wherein the mutant ORF2 polypeptide has a 5-GO production potential that is higher than that of WT ORF2, and wherein the mutant ORF2 polypeptide produces a lower % of CBG than WT ORF2. In some aspects, the 5-GO is produced ex vivo or in vitro. In some aspects, 5-GO is produced in cell-free systems. In some aspects, the 5-GO is produced in host cells, such as, for example, microbial host cells, such as, for example, bacteria, E. coli, yeast, microalgae; non-microbial hosts, such as, for example, insect cells, mammalian cells, plant cells; and whole terrestrial plants. In some aspects, the 5-GO is produced in the medium of host cell culture.
It is to be understood that the description above as well as the examples that follow are intended to illustrate, and not limit, the scope of the invention. Other aspects, advantages and modifications within the scope of the invention will be apparent to those skilled in the art to which the invention pertains.
All patents, patent applications, references, and journal articles cited in this disclosure are expressly incorporated herein by reference in their entireties for all purposes.
EXAMPLES Example 1: Methods for Generating and Studying ORF2 Variants A. Construction of a Synthesized Gene Library of n=96 Orf2 Variants with Select Amino Acid Substitutions and Other Orf2 Variants.
DNA plasmids encoding the 96 “tripleton” variants of orf2 (orf2 variants) were ordered and delivered in the background of the T5 expression vector pD441-SR from DNA2.0 (now ATUM, catalog pD441-SR). The sequences for the 96 variants are described as SEQ ID NO: DNA_150247-DNA_150342. Each Orf2 variant contains a unique combination of three amino acid substitutions relative to the base construct (SEQ ID NO: DNA_consensus).
All variants aside from the tripleton parental variants were created using site directed mutagenesis with QuikChange II Site-Directed Mutagenesis Kit (Agilent catalog #200523). Standard manufacturer protocols were employed.
B. Expression and Purification of Proteins from the Synthesized Orf2 Gene Library of Orf2 Variants.
DNA plasmids containing each of the Orf2 variants were individually transformed into OneShot BL21(DE3) chemically competent E. coli cells (Invitrogen catalog C600003) according to the chemically competent cell transformation protocol provided by Invitrogen. This resulted in 96 individual E. coli cell lines, each containing one plasmid encoding an Orf2 variant.
To induce protein expression, individual cell lines encoding each of the “orf2 variants” was individually inoculated into 2 milliliters LB media with 50 micrrograms per milliliter of Kanamycin sulfate in 15 milliliter culture tubes and grown at 37 degrees Celsius for 16 hours with vigorous shaking. After 16 hours, each culture was diluted into 38 milliliters LB media with 50 micrograms per milliliter of Kanamycin sulfate for a total of 40 milliliters. The absorbance at 600 nm (OD600) was monitored until it reached a value of 0.6 absorbance units. When the OD600 reached a value of 0.6, then IPTG was added to each culture to a final concentration of 500 micrograms per milliliter, resulting in an “induced culture.” Each “induced culture” was grown at 20 degrees Celsius with vigorous shaking for 20 hours.
After the cultures were grown under protein induction conditions, the target protein was extracted following a standard protein purification protocol. Each “induced culture” was spun at 4,000G for 5 minutes. The supernatant was discarded, leaving only a cell pellet. Each individual cell pellet was resuspended in 25 milliliters of a solution containing 20 millimolar Tris-HCL, 500 millimolar sodium chloride, 5 millimolar imidazole, and 10% glycerol (“lysis buffer”), resulting in a “cell slurry.” To each individual “cell slurry”, 30 microliters of 25 units per microliter Benzonase (Millipore, Benzonase, catalog number 70664-1), as well as 300 microliters of phosphatase and protease inhibitor (Thermo-Fisher, Halt Protease and Phosphatase Inhibitor Cocktail, EDTA-free, catalog number 78441) was added. Each individual “cell slurry” was then subjected to 30 second pulses of sonication, 4 times each, for a total of 120 seconds, using the Fisher Scientific Sonic Dismembrator Model 500 under 30% amplitude conditions. In between each 30 second pulse of sonication, the “cell slurry” was placed on ice for 30 seconds. After sonication, each individual “cell slurry” was centrifuged for 45 minutes at 14,000 times gravity.
Protein purification columns (Bio-Rad, Econo-Pac Chromotography Columns, catalog number 7321010) were prepared by adding 1.5 milliliters His60 resin slurry (Takara, His60 nickel superflow resin, catalog number 635660). 5 milliliters deionized water was added to resin slurry, to agitate and rinse the resin. The columns were then uncapped and the resulting flow-through was discarded. Then, 5 milliliters deionized water was added a second time, and the resulting flow-through was discarded. Then, 10 milliliters “lysis buffer” was added to the resin, completely disturbing the resin bed, and the flow-through was discarded.
The protein purification columns were capped, and the supernatant from the “cell slurry” was added to the resin bed without disturbing the resin bed. The columns were uncapped, allowing the supernatant to pass over the resin bed. The resin was then washed 2 times with 10 milliliters of a solution containing 20 millimolar Tris-HCl, 500 millimolar sodium chloride, and 20 millimolar imidazole (“wash buffer”). The flow-through from the wash steps was discarded. The protein was then eluted off the column with 10 milliliters of a solution containing 20 millimolar Tris-HCl, 200 millimolar sodium chloride, and 250 millimolar imidazole. The eluted protein was collected and dialyzed overnight in 4 liters of a solution containing 200 millimolar Tris-HCl and 800 millimolar sodium chloride in 3.5-5.0 kilodalton dialysis tubing (Spectrum Labs, Spectra/Por dialysis tubing, catalog number 133198). After overnight dialysis, protein was concentrated to approximately 10 milligrams per milliliter using centrifugal protein filters (Millipore Amicon Ultra-15 Ultracel 10K, catalog number UFC901024).
C. Screening of the Orf2 Protein Variants for Protein Activity and Phenotypes.
The library of orf2 variants were screened for protein expression by western blot with an anti-HIS antibody (Cell Signaling Technologies, anti-his monoclonal antibody, catalog number 23655) according to the protocol provided by Cell Signaling Technologies for the antibody. The variants that had detectable levels of protein expression as determined by western blot were used in a prenylation assay.
Proteins that exhibited detectable expression by Western blot were assayed for prenylation activity using a substrate (e.g. olivetolic acid, olivetol, divarinic acid, etc.) and a donor molecule (e.g. GPP, FPP, DMAPP, etc.). Unless otherwise stated, each prenylation reaction assay was performed in a volume of 20 microliters and contained 20 millimolar magnesium chloride (MgCl2), 2 millimolar donor molecule (e.g. GPP), 100 millimolar HEPES buffer at a pH of 7.5, 2 millimolar substrate (e.g. olivetolic acid), and 20 micrograms Orf2 protein or Orf2 variant protein. These reactions were incubated for 16 hours at 30° C.
For those prenylation reactions carried out at pH 5.3, the same prenylation reaction assay protocol above was carried out with the exception of replacing HEPES buffer with sodium citrate at pH 5.3.
To assess prenylation activity by HPLC the prenylation products were extracted from the assay reaction with the following protocol: 40 microliters of ethyl acetate was added to each reaction and vortexed thoroughly. After vortexing, each reaction was centrifuged for 10 minutes at 14,000G. The top layer (“organic layer”) was collected. This was repeated twice. The collected organic layer was evaporated, and the resulting residue was resuspended in 40 microliters of 100% methanol. After resuspending in methanol, 40 microliters of 100% HPLC grade water was added to bring the final solution to 50% methanol. These will be referred to as the “variant reactions with GPP.”
The final 50% methanol solutions were run on a Thermo Fisher UltiMate 3000 UHPLC with an Acclaim RSLC 120 angstrom C18 column with a 4 millimeter Phenomonex Securityguard guard column (54 millimeter total column length).
The wild type Orf2 standard prenylation assay produces two products as determined by HPLC, with retention times of approximately 6.9 minutes (an “early product” which is cannabigerolic acid [CBGA]) and 7.2 minutes (a “late product” 5-GOA) in a ratio of −20% CBGA to 80% 5-GOA.
Example 2: Generation of ORF2 Variants which Synthesize an Altered Amount of CBGA and/or 5-GOA, Compared to WT ORF2 A rational design approach was used to generate a library of 96 ORF2 triple mutants in which each triple mutant carried amino acid substitutions at 3 of 36 selected residues following the methods described in Example 1. These triple mutants may be interchangeably referred to as tripleton variants or tripleton mutants. Each amino acid substitution was employed 3-5 times in the library. From 66 of the 96 clones each carrying a unique tripleton ORF2 variant, ORF2 mutant proteins were expressed and their activity was analyzed as described in Example 1.
Table 1 provides a summary of the analysis performed on the enzymatic activity of the ORF2 variants to produce CBGA and 5-GOA using Olivetolic Acid (OA) as substrate and Geranyl pyrophosphate (GPP) as donor. Table 1 lists the mutations within each of the tripleton mutants as well the nMol of CBGA produced, nMol of 5-GOA produced, total prenylated products produced (nMol of CBGA+5-GOA), % CBGA within total prenylated products (nMol of CBGA/[nMol of CBGA+5-GOA]), % enzymatic activity (total prenylated products produced by a mutant/total prenylated products produced by wild-type ORF2), CBGA production (% CBGA among total prenylated products * % enzymatic activity), and %5-GOA within prenylated products (nMol of 5-GOA/[nMol of CBGA+5-GOA]) for each of the ORF2 variants.
TABLE 1
Analysis of ORF2 mutants and WT ORF2 based on production of CBGA from OA and GPP
CBGA
Production
Ser. nMol nMol Total % % (% CBGA %
No. CLONE Mutations CBGA 5-GOA Product CBGA Activity *% activity) 5-GOA
n/a WT WT 1.693073479 6.906720359 8.599793838 19.7% 100.0% 0.2 80.31%
1 F08 A53T_N173D_S214R 0.696655254 0.019606549 0.716261803 97.3% 8.6% 0.1 2.7%
2 G12 A17T_Q161W_A232S 6.109902555 0.614932664 6.724835219 90.9% 78.2% 0.7 9.1%
3 G11 S177W_Y288H_V294N 0.145430603 0.029376815 0.174807419 83.2% 2.0% 0.0 16.8%
5 H02 A53Q_S177W_L219F 3.42156966 1.080670716 4.502240376 76.0% 52.4% 0.4 24.0%
6 H06 V49L_E112D_G286E 0.070345009 0.025481912 0.095826921 73.4% 1.1% 0.0 26.6%
8 A04 L219F_V294N_Q295A 3.234527258 1.296738843 4.531266102 71.4% 40.6% 0.3 28.6%
10 C03 V49L_S214R_V271E 0.048617329 0.024425667 0.073042996 66.6% 0.8% 0.0 33.4%
11 F09 Q38G_D166E_Q295A 0.915880959 0.48336414 1.399245099 65.5% 16.8% 0.1 34.5%
13 H05 S177E_S214R_R228E 0.039715565 0.022115131 0.061830696 64.2% 0.7% 0.0 35.8%
14 G09 M106E_G205L_C209G 0.026257572 0.016437814 0.042695385 61.5% 0.5% 0.0 38.5%
15 G07 V49S_Y216A_V294N 0.503397419 0.325323475 0.828720894 60.7% 9.6% 0.1 39.3%
16 A11 V49L_D166E_L274V 0.028022123 0.018946396 0.046968518 59.7% 0.4% 0.0 40.3%
17 C10 A53Q_L274V_Q295A 0.157229392 0.108397148 0.26562654 59.2% 2.8% 0.0 40.8%
18 D11 F123H_L174V_S177E 0.573031341 0.397874307 0.970905647 59.0% 11.7% 0.1 41.0%
19 B11 V49S_K119D_F213M 0.029022913 0.020200687 0.049223599 59.0% 0.5% 0.0 41.0%
20 B10 M106E_Y121W_D166E 0.032314986 0.023237391 0.055552377 58.2% 0.6% 0.0 41.8%
21 G10 V49A_Y121W_C230S 0.0186463 0.013731186 0.032377485 57.6% 0.4% 0.0 42.4%
22 C05 A53Q_S177Y_Y288H 2.329839347 1.716266174 4.046105521 57.6% 42.6% 0.2 42.4%
23 C07 V49L_K119D_G205M 0.031761917 0.025613942 0.05737586 55.4% 0.6% 0.0 44.6%
24 A08 C25V_A232S_V271E 0.027785093 0.022445207 0.050230301 55.3% 0.5% 0.0 44.7%
25 C02 K118N_K119A_V271E 0.029312615 0.024755743 0.054068359 54.2% 0.6% 0.0 45.8%
26 D03 D227E_C230N_Q295W 0.28775349 0.249801954 0.537555444 53.5% 5.7% 0.0 46.5%
27 B05 A53Q_Y121W_A232S 0.01948907 0.017097967 0.036587037 53.3% 0.4% 0.0 46.7%
28 F10 K119D_Q161W_L298Q 0.044745852 0.039807235 0.084553087 52.9% 1.0% 0.0 47.1%
29 C06 Q161A_M162F_Q295A 0.926020543 0.852983892 1.779004435 52.1% 18.7% 0.1 47.9%
30 E09 A53T_M106E_Q161S 5.187648143 5.017560074 10.20520822 50.8% 122.8% 0.6 49.2%
31 A03 F123A_Y283L 0.051777719 0.05017164 0.101949359 50.8% 0.9% 0.0 49.2%
32 A10 V49S_K118Q_S177E 0.382933895 0.46547399 0.848407885 45.1% 7.6% 0.0 54.9%
33 A12 Y121W_S177Y_G286E 0.011983145 0.014853446 0.026836591 44.7% 0.2% 0.0 55.3%
34 B02 V49A_S177Y_C209G 0.025283118 0.031621336 0.056904454 44.4% 0.6% 0.0 55.6%
35 B06 D166E_S177Y_S214F 0.024782723 0.033997888 0.058780611 42.2% 0.6% 0.0 57.8%
36 H11 A108G_Q161S_G205M 2.068317092 2.915368365 4.983685458 41.5% 58.0% 0.2 58.5%
37 D10 V49A_F123A_Y288H 0.010034238 0.015117507 0.025151745 39.9% 0.3% 0.0 60.1%
38 A09 V49A_Q161S_V294A 3.196576244 4.907578558 8.104154803 39.4% 72.7% 0.3 60.6%
39 D04 A53T_D166E_Q295W 2.748485647 4.298125165 7.046610812 39.0% 74.2% 0.3 61.0%
40 E10 E112D_K119A_N173D 0.066631551 0.108199102 0.174830653 38.1% 2.1% 0.0 61.9%
41 F12 A17T_V49A_C230N 0.043402686 0.087998416 0.131401102 33.0% 1.5% 0.0 67.0%
42 C09 A108G_K119D_L298A 0.03605478 0.074993398 0.111048179 32.5% 1.2% 0.0 67.5%
43 H01 K119A_Q161A_R228Q 0.057018699 0.121006073 0.178024772 32.0% 2.1% 0.0 68.0%
44 H04 M162A_C209G_Y288H 0.061917303 0.154673884 0.216591187 28.6% 2.5% 0.0 71.4%
45 G06 K118Q_F123A_R228E 0.020437187 0.051491946 0.071929133 28.4% 0.8% 0.0 71.6%
46 B04 A53E_A108G_K118N 0.020779563 0.052812252 0.073591815 28.2% 0.8% 0.0 71.8%
47 D01 K118Q_Q161W_S214F 0.095601791 0.258515976 0.354117767 27.0% 3.7% 0.0 73.0%
48 A07 G205L_R228E_C230N 0.020700553 0.064562979 0.085263532 24.3% 0.8% 0.0 75.7%
49 D09 K118N_C209G_R228Q 0.008954438 0.028122524 0.037076962 24.2% 0.4% 0.0 75.8%
50 B09 C25V_F213M_Y216A 0.07951014 0.260364405 0.339874544 23.4% 3.6% 0.0 76.6%
51 G04 D227E_R228E_L298Q 0.015591256 0.056443095 0.072034351 21.6% 0.8% 0.0 78.4%
52 B08 K118Q_L174V_R228Q 0.044903871 0.167216794 0.212120666 21.2% 2.2% 0.0 78.8%
54 G05 A53T_K118N_S214F 0.581906769 2.595590177 3.177496945 18.3% 36.9% 0.1 81.7%
55 B12 A17T_F123W_L298A 0.125256782 0.602323739 0.727580521 17.2% 7.7% 0.0 82.8%
56 A05 A17T_C25V_E112G 0.964577298 6.867309216 7.831886514 12.3% 70.3% 0.1 87.7%
57 C08 V49S_S214G_V294A 0.390782196 3.025481912 3.416264108 11.4% 36.0% 0.0 88.6%
58 C11 E112D_L219F_V294F 0.971214116 7.558225508 8.529439625 11.4% 89.8% 0.1 88.6%
59 A02 Q38G_E112D_F123H 0.271319463 3.467916557 3.739236019 7.3% 33.5% 0.0 92.7%
60 D08 E112D_K119A_N173D 0.003976824 0.051557961 0.055534785 7.2% 0.7% 0.0 92.8%
61 D12 A53T_E112D_G205M 1.387332104 23.71798257 25.10531468 5.5% 302.1% 0.2 94.5%
62 G03 L219F_Y283L_L298W 0.286120622 5.898930552 6.185051173 4.6% 71.9% 0.0 95.4%
63 H10 M162A_N173D_S214F 0.189544377 3.951874835 4.141419212 4.6% 48.2% 0.0 95.4%
64 H09 E112G_G205M_L298W 0.422149065 13.32116451 13.74331358 3.1% 159.8% 0.0 96.9%
65 C01 V49S_M162A_Y283L 0.08967606 2.97900713 3.06868319 2.9% 32.3% 0.0 97.1%
66 D06 A53E_Q161A_V294N 0.453779299 16.8069052 17.2606845 2.6% 181.7% 0.0 97.4%
67 H07 F123A_M162F_S214G 0.049749802 1.903023501 1.952773304 2.5% 22.7% 0.0 97.5%
68 G02 A53E_F213M_R228Q 0.056544641 2.991418009 3.047962649 1.9% 35.4% 0.0 98.1%
69 C12 N173D_F213M_V294F 0.051988412 6.831726961 6.883715373 0.8% 72.4% 0.0 99.2%
70 D07 K119A_S214G_L298A 0.037266263 6.776340111 6.813606374 0.5% 82.0% 0.0 99.5%
The amount of CBGA or 5-GOA (in nMols) generated by each of the ORF2 triple mutant clones was measured using HPLC. FIG. 1 shows the total nMols of prenylated products generated using OA as substrate and GPP as donor by each of the ORF2 triple mutants, and the proportion of CBGA and 5-GOA within the total amount of prenylated products. An exemplary Wild Type ORF2 replicate is included in the graph for comparison purposes.
FIG. 2 shows the % CBGA within the total prenylated products produced by each of the ORF2 triple mutant clones using OA as substrate and GPP as donor. In this graph, the mutant clones are ordered based on decreasing % CBGA (from left to right) they produce, with the %5-GOA depicted in red. The black threshold line on the graph indicates the % CBGA that is produced by the wild type enzyme.
FIG. 3 shows the ORF2 enzymatic activity (using OA as substrate and GPP as donor) of each of the triple mutant ORF2 clones relative to the wild type enzyme. % activity was calculated by dividing the nMols of total prenylated products produced by a mutant by the nMols of total prenylated products produced by the wild type control, and expressed as a percentage. The red threshold line is the wild type Orf2% activity.
FIG. 4 shows the CBGA production potential of each of the ORF2 triple mutant clones when using OA as substrate and GPP as donor. CBGA production potential (interchangeably referred to herein as CBGA production quotient) represents the improvement in CBGA production vs. the wild type enzyme. CBGA production potential was calculated by multiplying the % CBGA by the % activity of each mutant. For instance, a wild type ORF2, which makes ˜20% CBGA, and has an activity of 100%, would have a CBGA Production Potential of 0.2. The red threshold line on the graph represents this wild type value of 0.2.
While the CBGA production potential analysis shown in FIG. 4 is useful to rank ORF2 mutant clones based on the amount of CBGA produced, such an analysis would not differentiate between a mutant that made 100% CBGA but was 20% as active as wild-type ORF2; or a mutant that made 10% CBGA and was 200% as active as wild type ORF2. Therefore, we employed a cluster analysis by plotting the CBGA Production Potential vs. %5-GOA (FIG. 5). %5-GOA was calculated in a similar manner as % CBGA. We used the top 16 mutants ranked based on their CBGA production potential for this analysis. High 5-GOA producing mutants cluster together towards the right of the graph and high CBGA producing mutants cluster towards the left of the graph.
Based on the analysis performed in FIG. 5, 13 mutants which cluster to the left of the graph were selected (Table 2). These clones were targeted for “breakdown” analysis. Breakdown analysis involves breaking a parent triple mutant into all pair wise doubleton combinations of mutations as well as all singleton mutations that make up the parental clone. For each parental clone targeted six unique mutants are generated (3 doubles and 3 singles).
TABLE 2
CBGA
Production Clone
Rank ID Mutations
1 G12 A17T_Q161W_A232S
2 E09 A53T_M106E_Q161S
3 H02 A53Q_S177W_L219F
4 A04 L219F_V294N_Q295A
5 D04 A53T_D166E_Q295W
6 A09 V49A_Q161S_V294A
7 C05 A53Q_S177Y_Y288H
8 H11 A108G_Q161S_G205M
10 F09 Q38G_D166E_Q295A
12 C06 Q161A_M162F_Q295A
14 F08 A53T_N173D_S214R
15 D11 F123H_L174V_S177E
16 G05 A53T_K118N_S214F
For the singleton and doubleton mutants resulting from the breakdown of triple mutants—E09, H02, A04, D04, A09, C05, H011, F09, D011, and G05—the total amount of prenylated products (and the respective proportion of CBGA and 5-GOA); and % CBGA within the prenylated products was calculated. FIGS. 6-15 depict the total amount of prenylated products and % CBGA produced using OA as substrate and GPP as donor for the mutants derived from E09 (FIG. 6); H02 (FIG. 7); A04 (FIG. 8); D04 (FIG. 9); A09 (FIG. 10); C05 (FIG. 11); H011 (FIG. 12); F09 (FIG. 13); D011 (FIG. 14); and G05 (FIG. 15). The % CBGA; % enzymatic activity; and the CBGA production potential for these clones, along with the mutations they carry, are listed in Table 3.
In a similar manner, the triple mutants, G12, C06 and F08, will also be subjected to “breakdown” analysis. Further, the singleton and double mutants resulting from the breakdown of G12, C06 and FO will be analyzed to determine the total amount of prenylated products (and the respective proportion of CBGA and 5-GOA); and % CBGA within the prenylated products produced by these mutants, as described above.
TABLE 3
Summary table of enzymatic activity of triple mutants; and the double and
single mutants resulting from the breakdown thereof
% CBGA
CBGA/5-G Production
nMol nMol Total (nMol % (% CBGA
CBGA 5GOA Product Calculation) Activity *% activity)
E9 A53T_M106E_Q161S 5.18764814 5.01756007 10.2052082 50.8% 122.8% 0.6
025 A53T_M106E 2.25762444 8.84592025 11.1035447 20.3% 163.7% 0.3
026 A53T_Q_161S 5.06402423 20.7614206 25.8254449 19.6% 190.3% 0.37
027 M106E_Q_161S 1.97260996 1.70913652 3.68174647 53.6% 27.1% 0.15
033 A53T 2.53858309 22.0230393 24.5616224 10.3% 228.1% 0.2
040 M106E 1.13455359 2.15698442 3.29153802 34.5% 30.6% 0.1
041 Q161S 5.90281801 11.3076974 17.2105154 34.3% 166.7% 0.6
H2 A53Q_S177W_L219F 3.42156966 1.08067072 4.50224038 76.0% 52.4% 0.4
007.2 A53Q_S177W 4.76897551 1.45933457 6.22831007 76.6% 57.8% 0.4
008 A53Q_L219F 1.86626284 9.95471349 11.8209763 15.8% 109.7% 0.2
009 S177W_L219F 1.5588886 0.40797465 1.96686325 79.3% 29.0% 0.2
032 A53Q 1.79288912 11.8606417 13.6535308 13.1% 126.8% 0.2
039.2 L219F 1.93410587 6.27917877 8.21328464 23.5% 76.3% 0.2
046 S177W 3.56125889 0.68108001 4.2423389 83.9% 41.1% 0.3
A4 L219F_V294N_Q295A 3.23452726 1.29673884 4.5312661 71.4% 40.6% 0.3
004 L219F_V294N 2.50545167 15.4131899 17.9186415 14.0% 166.3% 0.2
005.1 L219F_Q295A 2.25069792 0.63585952 2.88655744 78.0% 26.8% 0.2
006 V294N_Q295A 2.49960495 16.7207552 19.2203602 13.0% 178.4% 0.2
039.1 L219F 1.56952858 5.33575389 6.90528247 22.7% 64.1% 0.1
042 Q295A 3.49144061 1.05584896 4.54728957 76.8% 44.0% 0.3
050 V294N 2.00010535 12.8829548 14.8830602 13.4% 144.2% 0.2
D4 A53T_D166E_Q295W 2.74848565 4.29812517 7.04661081 39.0% 74.2% 0.3
016 A53T_D166E 0.82704767 12.2768022 13.1038499 6.3% 193.2% 0.1
017 A53T_Q295W 5.82552015 4.01524954 9.84076969 59.2% 145.1% 0.9
018 D166E_Q295W 3.43766131 1.92018748 5.3578488 64.2% 79.0% 0.5
033 A53T 2.53858309 22.0230393 24.5616224 10.3% 228.1% 0.2
034 D166E 0.93471161 4.71177713 5.64648875 16.6% 52.4% 0.1
043 Q295W 11.4932578 3.57948244 15.0727403 76.3% 146.0% 1.1
A9 V49A_Q161S_V294A 3.19657624 4.90757856 8.1041548 39.4% 72.7% 0.3
022 V49A_Q161S 1.74851198 2.17764721 3.9261592 44.5% 57.9% 0.3
023 V49A_V294A 0.78309191 2.6874835 3.47057541 22.6% 51.2% 0.1
024 Q161S_V294A 4.87521728 12.8399129 17.7151301 27.5% 261.2% 0.7
041 Q161S 5.90281801 11.3076974 17.2105154 34.3% 166.7% 0.6
049 V294A 2.4024493 16.2872987 18.689748 12.9% 181.0% 0.2
051 V49A 0.78854359 1.76458938 2.55313297 30.9% 24.7% 0.1
C5 A53Q_S177Y_Y288H 2.32983935 1.71626617 4.04610552 57.6% 42.6% 0.2
019 A53Q_S177Y 0.44303397 4.4004489 4.84348288 9.1% 71.4% 0.1
020 A53Q_Y288H 3.49228338 1.37476895 4.86705233 71.8% 71.8% 0.5
021 S177Y_Y288H 1.27124045 0.5814629 1.85270335 68.6% 27.3% 0.2
032 A53Q 1.79288912 11.8606417 13.6535308 13.1% 126.8% 0.2
047 S177Y 0.52881222 2.78901505 3.31782727 15.9% 32.1% 0.1
052 Y288H 2.15583355 0.63321891 2.78905246 77.3% 27.0% 0.2
H11 A108G_Q161S_G205M 2.06831709 2.91536837 4.98368546 41.5% 58.0% 0.2
010 A108G_Q161S 1.07729787 1.38302086 2.46031873 43.8% 36.3% 0.2
011 A108G_G205M 0.189202 1.1392263 1.3284283 14.2% 19.6% 0.0
012 Q161S_G205M 2.73157756 6.75105625 9.48263381 28.8% 139.8% 0.4
031 A108G 0.26987095 1.27515184 1.54502279 17.5% 14.4% 0.0
036 G205M 2.02009481 10.8182598 12.8383546 15.7% 119.2% 0.2
041 Q161S 5.90281801 11.3076974 17.2105154 34.3% 166.7% 0.6
F9 Q38G_D166E_Q295A 0.91588096 0.48336414 1.3992451 65.5% 16.8% 0.1
001 Q38G_D166E 0.44666842 2.19118035 2.63784878 16.9% 24.5% 0.0
002 Q38G_Q295A 2.20134317 7.38011619 9.58145935 23.0% 88.9% 0.2
003 D166E_Q295A 1.57469055 0.7702667 2.34495725 67.2% 21.8% 0.1
034 D166E 0.93471161 4.71177713 5.64648875 16.6% 52.4% 0.1
042 Q295A 3.49144061 1.05584896 4.54728957 76.8% 44.0% 0.3
044 Q38G 1.09681327 4.18807763 5.28489091 20.8% 51.2% 0.1
D11 F123H_L174V_S177E 0.57303134 0.39787431 0.97090565 59.0% 11.7% 0.1
013 F123H_L174V 0.1767448 1.15110906 1.32785386 13.3% 19.6% 0.0
014 F123H_S177E 0.51964709 0.37740956 0.89705665 57.9% 13.2% 0.1
015 L174V_S177E 4.34427179 1.80921574 6.15348753 70.6% 90.7% 0.6
035 F123H 0.28346063 2.26571165 2.54917227 11.1% 23.7% 0.0
038 L174V 1.91411641 6.27865065 8.19276705 23.4% 80.0% 0.2
045 S177E 6.41430076 2.15579614 8.57009691 74.8% 83.0% 0.6
G5 A53T_K118N_S214F 0.58190677 2.59559018 3.17749695 18.3% 36.9% 0.1
028 A53T_K118N 0.2830129 4.06093214 4.34394504 6.5% 64.1% 0.0
029.2 A53T_S214F 1.51042929 4.76392923 6.27435852 24.1% 92.5% 0.2
030 K118N_S214F 0.14572031 0.33747029 0.4831906 30.2% 7.1% 0.0
033 A53T 2.53858309 22.0230393 24.5616224 10.3% 228.1% 0.2
037 K118N 0.28854359 1.24042778 1.52897137 18.9% 14.2% 0.0
048 S214F 1.11524888 2.08588593 3.20113481 34.8% 31.0% 0.1
G12 A17T_Q161W_A232S 6.10990255 0.61493266 6.72483522 90.86% 78.20% 0.7
q423-1 A17T 1.06431393 5.17593082 6.24024475 17.06% 50.31% 0.1
EE09 Q161W 0.37682381 1.63856615 2.01538996 18.70% 19.52% 0.0
424-1 A232S 8.72878588 1.57004225 10.2988281 84.76% 83.03% 0.7
C06 Q161A_M162F_Q295A 0.92602054 0.85298389 1.77900443 52.05% 18.72% 0.10
DD01 Q161A 0.94174348 12.9347769 13.8765203 6.79% 192.32% 0.13
431-1 M162F 0.94661575 0.40150515 1.3481209 70.22% 10.87% 0.08
042 Q295A 3.49144061 1.05584896 4.54728957 76.78% 46.74% 0.36
F08 A53T_N173D_S214R 0.69665525 0.01960655 0.7162618 97.26% 8.62% 0.08
033 A53T 2.53858309 22.0230393 24.5616224 10.34% 220.33% 0.23
427-4 N173D 0.07008164 0.44725376 0.51733541 13.55% 4.17% 0.01
BB05 S214R 4.81951541 0.02554793 4.84506333 99.47% 49.81% 0.50
This analysis provided important insights into which positions on ORF2, when mutated, are likely to give rise to significant effects on the enzymatic activity of ORF2 in the reaction using Olivetolic Acid (OA) as substrate and Geranyl pyrophosphate (GPP) as donor. Based on this analysis, the following amino acid sites were selected for targeted amino acid site saturation mutagenesis—M106, Q161, 5177, Q295, Y288, 5214, M162 and A232.
Site saturated mutagenesis was done for Q295, Q161, 5214 and Y288 by replacing the wild type residue with each of the other 19 standard amino acids. The amount of total prenylated products, the CBGA production potential and GOA production potential was measured for each of the site saturated mutants. These results are depicted in FIGS. 16, 17, 18 and 97; and Tables 4, 5, 6 and 7.
TABLE 4
Q295 site saturated mutants
CBGA
% Production
CBGA/5-G potential 5-GOA
nMol nMol Total (nMol % (% CBGA* % 5- Production
Mutations CBGA 5GOA Product Calculation) Activity % activity) GOA Potential
Q295H 4.22220174 22.7255743 26.9477761 15.7% 277.0% 0.43 84.33% 2.34
Q295Y 2.00331841 8.06476102 10.0680794 19.9% 103.5% 0.21 80.10% 0.83
Q295D 4.84511456 7.65619224 12.5013068 38.8% 128.5% 0.50 61.24% 0.79
Q295R 0.10247564 4.60489834 4.70737398 2.2% 48.4% 0.01 97.82% 0.47
Q295W 11.4932578 3.57948244 15.0727403 76.3% 154.9% 1.18 23.75% 0.37
Q295N 1.44316566 2.64978875 4.09295441 35.3% 42.1% 0.15 64.74% 0.27
Q295G 5.17814064 2.21283338 7.39097401 70.1% 76.0% 0.53 29.94% 0.23
Q295S 1.59531209 1.62780565 3.22311774 49.5% 33.1% 0.16 50.50% 0.17
Q295K 0.44076903 1.42738315 1.86815218 23.6% 19.2% 0.05 76.41% 0.15
Q295C 5.74598367 1.17837338 6.92435705 83.0% 71.2% 0.59 17.02% 0.12
Q295M 13.1925467 1.1651043 14.3576511 91.9% 147.6% 1.36 8.11% 0.12
Q295P 1.31361601 1.10628466 2.41990067 54.3% 24.9% 0.14 45.72% 0.11
Q295A 3.49144061 1.05584896 4.54728957 76.8% 46.7% 0.36 23.22% 0.11
Q295L 11.1453516 0.5400713 11.6854229 95.4% 120.1% 1.15 4.62% 0.06
Q2951 6.45699236 0.44982836 6.90682072 93.5% 71.0% 0.66 6.51% 0.05
Q295F 16.5020543 0.44533932 16.9473936 97.4% 174.2% 1.70 2.63% 0.05
Q295E 1.93961022 0.39358331 2.33319353 83.1% 24.0% 0.20 16.87% 0.04
Q295T 0.605399 0.37734354 0.98274254 61.6% 10.1% 0.06 38.40% 0.04
Q295V 5.16273374 0.35549247 5.51822621 93.6% 56.7% 0.53 6.44% 0.04
TABLE 5
Q161 site saturated mutants
CBGA
Production
potential 5-GOA
nMol nMol Total % % (% CBGA* % Production
Mutations CBGA 5GOA Product CBGA Activity % Activity) 5-GOA potential
Q161C 1.254227021 11.51921 12.77343748 9.82% 208.10% 0.2 90.18% 1.88
Q161A 0.941743482 12.934777 13.87652035 6.79% 192.32% 0.1 93.21% 1.79
Q161L 4.847326837 9.7688144 14.6161412 33.16% 238.12% 0.8 66.84% 1.59
Q161T 3.438504082 9.1266174 12.56512146 27.37% 204.71% 0.6 72.63% 1.49
Q161P 0.405504346 14.929496 15.33499999 2.64% 148.54% 0.0 97.36% 1.45
Q1611 2.557940479 8.7985213 11.35646174 22.52% 185.01% 0.4 77.48% 1.43
Q161S 4.329707664 8.0136652 12.34337283 35.08% 201.09% 0.7 64.92% 1.31
Q161F 7.260284435 4.7907314 12.05101588 60.25% 196.33% 1.2 39.75% 0.78
Q161V 1.213141954 7.0618564 8.274998305 14.66% 0.667157399 0.1 85.34% 0.57
Q161N 1.500052673 4.9663322 6.466384862 23.20% 70.58% 0.2 76.80% 0.54
Q161G 1.023755597 4.8859916 5.909747147 17.32% 64.50% 0.1 82.68% 0.53
Q161M 2.039636555 3.1520993 5.191735842 39.29% 84.58% 0.3 60.71% 0.51
Q161K 1.740242297 1.3897544 3.12999672 55.60% 43.38% 0.2 44.40% 0.19
Q161W 0.376823808 1.6385661 2.015389956 18.70% 19.52% 0.0 81.30% 0.16
Q161Y 16.66784303 0.853644 17.52148708 95.13% 285.45% 2.7 4.87% 0.14
Q161R 12.29307348 0.8494851 13.14255856 93.54% 214.11% 2.0 6.46% 0.14
Q161E 0.711877798 1.1597571 1.871634862 38.04% 0.150897318 0.1 61.96% 0.09
Q161D 1.577482223 0.6375099 2.214992125 71.22% 21.46% 0.2 28.78% 0.06
Q161H 28.94774822 0.1498548 29.09760299 99.48% 474.04% 4.7 0.52% 0.02
TABLE 6
S214 site saturated mutants
CBGA
Production
potential 5-GOA
nMol nMol Total % % (% CBGA* % Production
Mutations CBGA 5GOA Product CBGA Activity % Activity) 5-GOA potential
S214G 0.24445615 20.3160153 20.5604715 1.19% 1.93572414 0.0 98.81% 1.91
S214A 0.25283118 10.2950885 10.5479196 2.40% 0.85040774 0.0 97.60% 0.83
S214T 1.76771135 6.74755743 8.51526878 0.20759314 87.5% 0.2 79.24% 0.69
S214V 1.32546747 5.81905202 7.14451949 0.18552227 73.4% 0.1 81.45% 0.60
S214F 0.49836713 1.06515712 1.56352425 0.31874602 16.1% 0.1 68.13% 0.11
S214C 0.10466157 1.04436229 1.14902387 0.09108738 11.8% 0.0 90.89% 0.11
S214N 0.68001053 0.82479535 1.50480589 0.45189253 15.5% 0.1 54.81% 0.08
S214D 0.45604425 0.81383681 1.26988106 0.35912359 13.1% 0.0 64.09% 0.08
S214W 0.04042665 0.43999208 0.48041873 0.08414879 4.9% 0.0 91.59% 0.05
S214I 0.56523571 0.41107737 0.97631308 0.57894924 10.0% 0.1 42.11% 0.04
S214Y 0.13197261 0.2728413 0.40481391 0.32600809 4.2% 0.0 67.40% 0.03
S214M 0.32538846 0.17685503 0.50224349 0.64786994 5.2% 0.0 35.21% 0.02
S214K 0.3499605 0.14536572 0.49532622 0.70652528 5.1% 0.0 29.35% 0.01
S214Q 9.75167237 0.17533668 9.92700905 98.23% 0.8003479 0.8 1.77% 0.01
S214E 0.92546747 0.1245709 1.05003837 0.88136538 10.8% 0.1 11.86% 0.01
S214L 0.09649723 0.09704251 0.19353975 0.4985913 2.0% 0.0 50.14% 0.01
S214P 0.00397682 0.07123053 0.07520735 5.29% 0.00734429 0.0 94.71% 0.01
S214R 4.81951541 0.02554793 4.84506333 0.99472702 49.8% 0.5 0.53% 0.00
S214H 11.2844614 0.02310536 11.3075668 0.99795665 116.2% 1.2 0.20% 0.00
TABLE 7
Y288 site saturated mutants
CBGA
Production
potential 5-GOA
nMol nMol 5- Total % % (% CBGA* % 5- Production
Mutations CBGA GOA Product Activity CBGA % Activity) GOA potential
Y288A 18.58056 0.080539 18.6611 150.45% 99.57% 1.50 0.43% 0.65%
Y288C 15.67369 0.122326 15.79602 127.35% 99.23% 1.26 0.77% 0.99%
Y288D 0.84335 0.022907 0.866257 6.98% 97.36% 0.07 2.64% 0.18%
Y288E 0.689123 0.020267 0.70939 5.72% 97.14% 0.06 2.86% 0.16%
Y288F 17.65373 0.056641 17.71037 142.79% 99.68% 1.42 0.32% 0.46%
Y288G 3.125836 0.044626 3.170463 25.56% 98.59% 0.25 1.41% 0.36%
Y288H 1.618251 0.489636 2.107887 16.99% 76.77% 0.13 23.23% 3.95%
Y288I 18.85518 0.210655 19.06583 153.71% 98.90% 1.52 1.10% 1.70%
Y288K 0.169213 0.019144 0.188357 1.52% 89.84% 0.01 10.16% 0.15%
Y288L 22.29837 0.154872 22.45324 181.03% 99.31% 1.80 0.69% 1.25%
Y288M 14.31317 0.177779 14.49095 116.83% 98.77% 1.15 1.23% 1.43%
Y288P 20.40645 0.068656 20.47511 165.08% 99.66% 1.65 0.34% 0.55%
Y288R 0.299368 0.050568 0.349936 2.82% 85.55% 0.02 14.45% 0.41%
Y288S 3.383408 0.064497 3.447905 27.80% 98.13% 0.27 1.87% 0.52%
Y288T 14.22997 0.21356 14.44353 116.45% 98.52% 1.15 1.48% 1.72%
Y288W 4.307032 0.107671 4.414703 35.59% 97.56% 0.35 2.44% 0.87%
Y288N 1.164077 0.105625 1.269701 10.24% 91.68% 0.09 8.32% 0.85%
Similarly, site saturated mutagenesis will also be completed for M106, and S177; and the amount of total prenylated products and the CBGA production potential will be measured for each of these site saturated mutants.
From the results described above, multiple mutations of Q295, Q161, Y288 and 5214 that have significantly higher CBGA production potential and/or the total amount of prenylated products, as compared to WT ORF2, were identified. Thus, the ORF2 mutants disclosed herein have unexpectedly superior enzymatic functions, in a reaction using OA as a substrate and GPP as donor, as compared to WT ORF2.
Finally, ORF2 stacking mutants, that carry different novel combinations of the mutations identified by our analysis as being important for ORF2's enzymatic activity, were analyzed to determine the total amount of prenylated products they produce (FIG. 19); % enzymatic activity (FIG. 20), % CBGA (FIG. 21), and CBGA production potential (FIG. 22). Table 8 provides a summary of the enzymatic function, using OA as substrate and GPP as donor, for each of the stacking mutants. The analysis of the stacking mutants shows that multiple stacking mutants have significantly higher % enzymatic activity, % CBGA, and CBGA production potential, compared to the WT ORF2 or either singleton substitution variant on its own, thereby indicating that the ORF2 stacking mutants disclosed herein have synergistically enhanced effects compared to the individual single mutants. Thus, the ORF2 stacking mutants disclosed herein have unexpectedly superior enzymatic functions, in a reaction using OA and GPP, as compared to WT ORF2.
For instance, ORF2 double mutants, A53T-S214R; Q161S-Q295L; Q161S-Q295F; S177W-Q295A have synergistically enhanced CBGA production potential and % activity as compared to either of the single mutants. See FIGS. 82-85.
More stacking mutants will be generated as described above, based on the breakdown analysis of triple mutants, G12, C06 and F08, and planned site saturation mutagenesis experiments described above. These stacking mutants will further be analyzed to determine their % enzymatic activity, % CBGA, %5-GOA and CBGA production potential.
TABLE 8
CBGA
Production
nMol nMol 5- Total % % (% CBGA* %
Mutations OA CBGA 5-GOA CBGA GOA Product Activity CBGA % Activity) 5-GOA
A53T_S214R 30.1449 62.6579 0.1069 16.5019489 0.07057037 16.5725193 229.69% 99.57% 2.29 0.43%
Q161S_Q295L 29.7788 57.0803 0.4619 15.0329997 0.30492474 15.3379245 212.58% 98.01% 2.08 1.99%
Q161S_Q295F 29.5883 55.4403 0.6309 14.6010798 0.41649063 15.0175704 208.14% 97.23% 2.02 2.77%
A53T_Q295F 30.0898 52.6928 1.1825 13.8774822 0.78063111 14.6581133 203.16% 94.67% 1.92 5.33%
A53T_Q161S_Q295W 32.3733 68.7659 4.4902 18.1105873 2.9642197 21.074807 157.59% 85.93% 1.35 14.07%
Q161S_V294A_Q295W 39.2778 68.3285 2.8122 17.9953911 1.8564827 19.8518738 148.45% 90.65% 1.35 9.35%
S214R_Q295F 43.7129 34.6447 0.1255 9.12422965 0.08284922 9.20707888 127.61% 99.10% 1.26 0.90%
A53T_Q161S_ 33.6064 63.9702 4.908 16.8475639 3.24003169 20.0875956 150.21% 83.87% 1.26 16.13%
V294A_Q295W
A53T_Q161S_ 36.0016 58.018 7.059 15.2799579 4.66002112 19.939979 149.11% 76.63% 1.14 23.37%
V294N_Q295W
S177W_Q295A 39.381 56.035 0.1016 14.7577035 0.06707156 14.824775 110.86% 99.55% 1.10 0.45%
A53T_S177W 54.8833 25.6741 1.8121 6.76168027 1.19626353 7.95794381 110.29% 84.97% 0.94 15.03%
Q161S_V294A_Q295A 37.0068 46.9066 9.5266 12.3535949 6.28901505 18.64261 139.41% 66.27% 0.92 33.73%
Q16S_S214R 58.7858 24.2694 0.1169 6.39173031 0.0771719 6.46890222 89.66% 98.81% 0.89 1.19%
A53T_Q295W 44.9864 41.0901 9.6799 10.8217277 6.39021653 17.2119442 128.71% 62.87% 0.81 37.13%
Q161S_S177W 54.4823 21.9926 0.6705 5.79209903 0.44263269 6.23473172 86.41% 92.90% 0.80 7.10%
A53T_V294A_Q295W 48.2017 38.5309 12.362 10.1477219 8.16081331 18.3085352 136.91% 55.43% 0.76 44.57%
A53T_V294A_Q295A 50.4412 37.8163 4.0888 9.95952067 2.69923422 12.6587549 94.66% 78.68% 0.74 21.32%
A53T_Q161S_ 29.5636 36.8247 18.7595 9.69836713 12.3841431 22.0825103 165.13% 43.92% 0.73 56.08%
V294A_Q295A
A53T_Q161S_Q295A 54.602 34.1016 14.121 8.98119568 9.32202271 18.3032184 136.87% 49.07% 0.67 50.93%
A53T_A161S 31.9028 16.0001 24.5577 4.21387938 16.2118431 20.4257225 283.09% 20.63% 0.58 79.37%
A53T_Q161S_ 51.4284 28.9549 15.8073 7.62573084 10.435239 18.0609698 135.06% 42.22% 0.57 57.78%
V294N_Q295A
S177W_S214R 73.1056 14.1529 0.0913 3.72739004 0.06027198 3.78766203 52.50% 98.41% 0.52 1.59%
A53T_Q295A 70.1304 26.0366 3.7553 6.85715038 2.47907314 9.33622353 69.81% 73.45% 0.51 26.55%
A53T_Q161S_V294A 43.4482 15.381 33.5956 4.0508296 22.1782414 26.229071 196.14% 15.44% 0.30 84.56%
A53T_Q161S_V294N 51.6804 11.9604 27.5382 3.1499605 18.1794296 21.3293901 159.50% 14.77% 0.24 85.23%
V49A_S214R 91.0858 3.4446 0.0741 0.90718989 0.04891735 0.95610724 13.25% 94.88% 0.13 5.12%
A53T_V294A 37.5911 5.8761 36.6497 1.54756387 24.1944151 25.741979 192.49% 6.01% 0.12 93.99%
V49A_Q295L 81.26 2.3805 0.277 0.62694232 0.18286242 0.80980475 11.22% 77.42% 0.09 22.58%
Example 3: Generation of ORF2 Variants with Improved Enzymatic Function Using OA as Substrate and GPP as Donor Four triple mutants (D12 carrying A53T_E112D_G205M mutations; D6 carrying A53E_Q161A_V294N mutations; H9 carrying E112G_G205M_L298W mutations; and E9 carrying A53T_M106E_Q161S mutations) that had improved activity vs. the WT ORF2, based on FIG. 3 were analyzed further. A53 (which was present in 3 out of 4 clones) and E112, Q161, and G205 (which were present in 2 out of 4 clones) were identified as target residues for site saturation.
Site-saturated mutagenesis was done for A53 by replacing the wild type residue at this site with each of the 19 other standard amino acids. % of enzymatic activity as compared to WT ORF2 was measured. These results are depicted in FIG. 23 and Table 9.
The site saturated mutants of Q161 described in Example 1 were also analyzed to determine their % activity in a reaction using OA as substrate and GPP as donor. These results are depicted in FIG. 24 and Table 10.
From the results described above, multiple mutations of A53 and Q161 that have significantly higher % activity, as compared to WT ORF2, were identified. Thus, the ORF2 mutants disclosed herein have unexpectedly superior enzymatic functions, in a reaction using OA as a substrate and GPP as donor, as compared to WT ORF2.
Similarly, site saturated mutagenesis will also be completed for E112 and G205 as described above; and % of enzymatic activity, as compared to WT ORF2, will be measured for each of these mutants.
TABLE 9
% activity of A53 site saturation mutants
nMol nMol Total %
Mutations CBGA 5-GOA Product Activity
A53T 2.38941269 19.8774096 22.2668223 228.9%
A53I 3.24627337 17.3253235 20.5715968 211.5%
A53V 3.89741902 15.1585688 19.0559878 195.9%
A53L 3.44464051 14.9749802 18.4196207 189.3%
A53M 2.57000263 12.7041194 15.274122 157.0%
A53K 1.96415591 13.2969369 15.2610928 156.9%
A53S 2.15754543 11.8968841 14.0544295 144.5%
A53Q 1.67719252 10.5953921 12.2725847 126.2%
A53N 0.64851198 6.83212305 7.48063504 76.9%
A53R 0.86465631 5.83100079 6.6956571 68.8%
A53W 0.51358968 5.14932664 5.66291632 58.2%
A53G 0.72675797 3.94533932 4.67209729 48.0%
A53D 0.77882539 3.02079482 3.79962021 39.1%
A53Y 0.19694496 0.82981252 1.02675747 10.6%
A53H 0.10524098 0.3159493 0.42119028 4.3%
A53F 0.14669476 0.12793768 0.27463244 2.8%
A53E 0.0294443 0.16054925 0.18999355 2.0%
A53P 0.0118778 0.25745973 0.26933753 0.02630185
TABLE 10
% activity of Q161 site saturation mutants
nMol nMol 5- Total % % CBGA Production
Mutations CBGA GOA Product CBGA Activity (% CBGA* % Activity)
Q161Y 16.667843 0.85364405 17.5214871 0.95128016 285.5% 2.7
Q161R 12.2930735 0.84948508 13.1425586 0.9353638 214.1% 2.0
Q161D 21.4621807 0.13090837 21.593089 0.99393749 209.2% 2.1
Q161C 1.25422702 11.5192105 12.7734375 0.09819025 208.1% 0.2
Q161T 3.43850408 9.12661738 12.5651215 0.27365466 204.7% 0.6
Q161S 4.32970766 8.01366517 12.3433728 0.35077185 201.1% 0.7
Q161F 7.26028444 4.79073145 12.0510159 0.60246244 196.3% 1.2
Q161A 0.94174348 12.9347769 13.8765203 0.06786597 192.3% 0.1
Q1611 2.55794048 8.79852126 11.3564617 0.22524097 185.0% 0.4
Q161P 0.40550435 14.9294956 15.335 0.02644306 148.5% 0.0
Q161M 2.03963656 3.15209929 5.19173584 0.39286216 84.6% 0.3
Q161Q 1.44664209 5.77105889 7.21770097 0.20042976 78.8% 0.2
Q161N 1.50005267 4.96633219 6.46638486 0.23197702 70.6% 0.2
Q161G 1.0237556 4.88599155 5.90974715 0.17323171 64.5% 0.1
Q161K 1.7402423 1.38975442 3.12999672 0.55598854 43.4% 0.2
Q161W 0.37682381 1.63856615 2.01538996 0.18697315 19.5% 0.0
Example 4: Generation of ORF2 Variants which Synthesize an Altered Amount of CBGVA and/or 5-GDVA Compared to WT ORF2 The library of tripleton mutants described in Example 1, was screened for the ability to produce CBGVA and 5-GDVA using DVA as substrate and Geranyl pyrophosphate (GPP) as donor; and this enzymatic activity was compared to that of the WT ORF2. Table 5 provides the summary of the analysis performed on the enzymatic activity of the ORF2 triple mutants to produce CBGVA and 5-GDVA using DVA as substrate and Geranyl pyrophosphate (GPP) as donor. Table 11 lists the mutations within each of the tripleton mutants as well the nMol CBGVA, nMol of 5-GDVA, total prenylated products, % CBGVA products among prenylated products, % enzymatic activity, CBGVA production potential and %5-GDVA among total prenylated products of each of the ORF2 triple mutants.
TABLE 11
CBGVA
CLONE nMol nMol 5- Total % % Production %
ID Mutations CBGVA GDVA Products CBGVA Activity Potential 5-GOA
G12 A17T_Q161W_A232S 15.44563535 1.780798893 17.22643424 89.66% 202.28% 1.81 10.3375943
A09 V49A_Q161S_V294A 13.27689212 2.418751236 15.69564336 84.59% 206.78% 1.75 15.4103351
H03 A17T_F213M_S214R 4.132448537 0.488308286 4.620756823 89.43% 67.42% 0.60 10.5677123
H02 A53Q_S177W_L219F 3.203799721 2.079716235 5.283515956 60.64% 62.04% 0.38 39.3623536
C05 A53Q_S177Y_Y288H 2.742222566 0.457534111 3.199756677 85.70% 42.15% 0.36 14.2990282
E09 A53T_M106E_Q161S 3.030297168 7.080383627 10.11068079 29.97% 118.73% 0.36 70.0287525
F08 A53T_N173D_S214R 2.498181396 1.595881946 4.094063342 61.02% 48.08% 0.29 38.9803922
D04 A53T_D166E_Q295W 1.938244854 2.469102235 4.407347088 43.98% 58.06% 0.26 56.0224141
H11 A108G_Q161S_G205M 2.059394831 2.31374827 4.3731431 47.09% 51.35% 0.24 52.9081308
D06 A53E_Q161A_V294N 1.045658567 3.092891042 4.138549609 25.27% 54.52% 0.14 74.733695
A10 V49S_K118Q_S177E 0.813341588 0.932123789 1.745465377 46.60% 23.00% 0.11 53.402594
G07 V49S_Y216A_V294N 0.898003405 0.146628436 1.044631841 85.96% 12.27% 0.11 14.0363744
C11 El 12D L219F_V294F 0.771126761 5.599490805 6.370617565 12.10% 83.93% 0.10 87.8955729
C06 Q161A_M162F_Q295A 0.737424547 0.764830927 1.502255475 49.09% 19.79% 0.10 50.9121744
H08 M106E_M162A_Y216A 0.768650364 0.224120032 0.992770395 77.42% 11.41% 0.09 22.575213
A04 L219F_V294N_Q295A 0.652569262 3.246737196 3.899306458 16.74% 51.37% 0.09 83.2644787
A05 A17T_C25V_E112G 0.648467729 6.486380265 7.134847994 9.09% 94.00% 0.09 90.9112608
C08 V49S_S214G_V294A 0.547012846 0.777733834 1.324746681 41.29% 17.45% 0.07 58.7081172
D12 A53T_E112D_G205M 0.548754063 11.45797904 12.0067331 4.57% 140.99% 0.06 95.4296139
H10 M162A_N173D_S214F 0.437741836 1.19524916 1.632990995 26.81% 19.18% 0.05 73.193861
H09 E112G_G205M_L298W 0.332843213 3.759738976 4.092582189 8.13% 48.06% 0.04 91.8671587
G05 A53T_K118N_S214F 0.321157716 0.749060708 1.070218423 30.01% 12.57% 0.04 69.991386
F09 Q38G_D166E_Q295A 0.295774648 0.495624876 0.791399524 37.37% 9.29% 0.03 62.6263804
D11 F123H_L174V_S177E 0.215059588 0.421939885 0.636999474 33.76% 7.48% 0.03 66.2386553
B09 C25V_F213M_Y216A 0.166228138 0.315379672 0.48160781 34.52% 6.34% 0.02 65.4847503
C01 V49S_M162A_Y283L 0.139142548 1.386320941 1.525463489 9.12% 20.10% 0.02 90.8786707
D07 K119A_S214G_L298A 0.128347005 0.796470239 0.924817244 13.88% 10.86% 0.02 86.1219062
G03 L219F_Y283L_L298W 0.1271475 1.259392921 1.386540421 9.17% 16.28% 0.01 90.8298742
H01 K119A_Q161A_R228Q 0.125715833 0.073017599 0.198733433 63.26% 2.33% 0.01 36.7414774
C12 N173D_F213M_V294F 0.109038848 6.163140202 6.27217905 1.74% 82.63% 0.01 98.2615476
D03 D227E_C230N_Q295W 0.079863798 0.046247775 0.126111574 63.33% 1.66% 0.01 36.6721103
A02 Q38G_E112D_F123H 0.078509519 0.064119043 0.142628562 55.04% 1.88% 0.01 44.9552616
G02 A53E_F213M_R228Q 0.063728525 3.011123196 3.074851721 2.07% 44.86% 0.01 97.9274277
A03 V49L_F123A_Y283L 0.079089924 0.065948191 0.145038115 54.53% 1.70% 0.01 45.4695586
G04 D227E_R228E_L298Q 0.064231543 0.496761914 0.560993457 11.45% 6.59% 0.01 88.5503935
A03 V49L_F123A_Y283L 0.054867668 0.61311054 0.667978207 8.21% 8.80% 0.01 91.7860093
B08 K118Q_L174V_R228Q 0.041363566 0.565651572 0.607015138 6.81% 8.00% 0.01 93.1857439
E10 E112D_K119A_N173D 0.041595728 0.173002768 0.214598497 19.38% 2.52% 0.00 80.6169527
F12 A17T_V49A_C230N 0.026234329 0.084017204 0.110251533 23.79% 1.29% 0.00 76.2050211
D09 K118N_C209G_R228Q 0.023951401 0.015300573 0.039251974 61.02% 0.46% 0.00 38.9803922
A08 C25V_A232S_V271E 0.016019192 0.027906862 0.043926054 36.47% 0.58% 0.00 63.5314565
D10 V49A_F123A_Y288H 0.015825724 0.010109749 0.025935472 61.02% 0.30% 0.00 38.9803922
E08 F123H_L274V_L298A 0.010021668 0.090493376 0.100515044 9.97% 1.69% 0.00 90.029683
D08 El 12D_K119A_N173D 0.009557344 0.066590864 0.076148208 12.55% 0.89% 0.00 87.449023
G11 S177W_Y288H_V294N 0.009479957 0.008305319 0.017785276 53.30% 0.21% 0.00 46.6977254
D01 K118Q_Q161W_S214F 0.007080947 0.458745304 0.465826251 1.52% 6.14% 0.00 98.4799167
H07 F123A_M162F_S214G 0.007080947 0.046099466 0.053180413 13.31% 0.62% 0.00 86.6850467
H06 V49L_E112D_G286E 0.006229686 0.006896381 0.013126067 47.46% 0.15% 0.00 52.5395857
H05 S177E_S214R_R228E 0.006152298 0.008503065 0.014655363 41.98% 0.17% 0.00 58.0201581
F10 K119D_Q161W_L298Q 0.006074911 0.032603322 0.038678233 15.71% 0.45% 0.00 84.2937215
A11 V49L_D166E_L274V 0.004527163 0.018316195 0.022843358 19.82% 0.30% 0.00 80.1817101
G08 F123W_M162F_C209G 0.005068875 0.015943247 0.021012122 24.12% 0.25% 0.00 75.8764258
B04 A53E_A108G_K118N 0.003714595 0.042193989 0.045908584 8.09% 0.60% 0.00 91.908713
C10 A53Q_L274V_Q295A 0.003675902 0.187685387 0.191361288 1.92% 2.52% 0.00 98.0790777
G10 V49A_Y121W_C230S 0.003985451 0.011073759 0.01505921 26.47% 0.18% 0.00 73.5347931
G09 M106E_G205L_C209G 0.003908064 0.011123196 0.015031259 26.00% 0.18% 0.00 74.0004235
G06 K118Q_F123A_R228E 0.003288965 0.027288906 0.030577871 10.76% 0.36% 0.00 89.2439714
H04 M162A_C209G_Y288H 0.003095496 0.014435436 0.017530932 17.66% 0.21% 0.00 82.3426613
C07 V49L_K119D_G205M 0.002515091 0.031491003 0.034006093 7.40% 0.45% 0.00 92.6040003
A07 G205L_R228E_C230N 0.001818604 0.101270516 0.10308912 1.76% 1.36% 0.00 98.2358915
C03 V49L_S214R_V271E 0.001625135 0.030205656 0.031830791 5.11% 0.42% 0.00 94.8944548
G08 F123W_M162F_C209G 0.00177991 0.035174016 0.036953926 4.82% 0.44% 0.00 95.1834341
B06 D166E_S177Y_S214F 0.001392973 0.030823611 0.032216584 4.32% 0.42% 0.00 95.6762231
B05 A53Q_Y121W_A232S 0.001392973 0.031688748 0.033081721 4.21% 0.44% 0.00 95.7892964
B02 V49A_S177Y_C209G 0.001083424 0.039351394 0.040434818 2.68% 0.53% 0.00 97.3205676
B10 M106E_Y121W_D166E 0.000812568 0.027807989 0.028620557 2.84% 0.38% 0.00 97.1608948
B11 V49S_K119D_F213M 0.000773874 0.028722563 0.029496437 2.62% 0.39% 0.00 97.3763814
A12 Y121W_S177Y_G286E 0.000464324 0.031589875 0.0320542 1.45% 0.42% 0.00 98.5514397
C09 A108G_K119D_L298A 0.000348243 0.03512458 0.035472823 0.98% 0.47% 0.00 99.0182814
C02 K118N_K119A_V271E 0.000116081 0.018711687 0.018827768 0.62% 0.25% 0.00 99.383458
The amount of CBGVA or 5-GDVA (in nMols) generated by each of the ORF2 triple mutant clones was measured using HPLC. FIG. 25 shows the total nMols of prenylated products generated using DVA as substrate and GPP as donor by each of the ORF2 triple mutants, and the proportion of CBGVA and 5-GDVA within the total prenylated products. An exemplary wild type ORF2 replicate is included in the graph for comparison purposes.
FIG. 26 shows the % CBGVA and %5-GDVA within the total prenylated products produced by each of the ORF2 mutant clones, using DVA as substrate and GPP as donor. In this graph, the mutant clones are ordered based on decreasing % CBGA (from left to right) they produce, with the %5-GDVA depicted in red. The black threshold line on the graph indicates the % CBGVA that is produced by the wild type enzyme.
FIG. 27 shows the % ORF2 enzymatic activity (using DVA as substrate and GPP as donor) of each of the mutant ORF2 clones relative to the wild type enzyme. % activity was calculated by dividing the nMols of total prenylated products produced by a mutant by the nMols of total prenylated products produced by the wild type control, and expressed as a percentage. The red threshold line represents the wild type Orf2% activity.
FIG. 28 shows the CBGVA production potential of each of the ORF2 mutant clones when using DVA as substrate and GPP as donor. CBGVA production potential (interchangeably referred to herein as CBGVA production quotient) represents the improvement in CBGVA production vs. the wild type enzyme. CBGVA production potential was calculated by multiplying the % CBGVA of the Total Products by the % Activity of each mutant. The red threshold line on the graph represents this wild type value.
While the CBGVA production potential analysis shown in FIG. 28 is useful to rank ORF2 mutant clones based on the amount of CBGVA produced, such an analysis would not differentiate between a mutant that made 100% CBGVA but was 20% as active as wildtype ORF2; or a mutant that made 10% CBGVA and was 200% as active as wild type ORF2. Therefore, a cluster analysis was employed by plotting the CBGVA Production Potential vs. %5-GDVA (FIG. 29). %5-GDVA was calculated in a similar manner as % CBGVA. We used the top 25 mutants ranked based on their CBGVA production potential for this analysis. High 5-GDVA producing mutants cluster together towards the right of the graph and high CBGVA producing mutants cluster towards the left of the graph.
Based on the analysis performed in FIG. 29, 24 mutants which cluster to the left of the graph were selected (Table 12). These clones were targeted for “breakdown” analysis.
TABLE 12
CBGVA Targeted
Production Clone for
Rank ID Mutations Breakdown
1 G12 A17T_Q161W_A232S YES
2 A09 V49A_Q161S_V294A YES
3 H03 A17T_F213M_S214R YES
4 H02 A53Q_S177W_L219F YES
5 C05 A53Q_S177Y_Y288H YES
6 E09 A53T_M106E_Q161S YES
7 F08 A53T_N173D_S214R YES
8 D04 A53T_D166E_Q295W YES
9 H11 A108G_Q1615_G205M YES
10 D06 A53E_Q161A_V294N YES
11 A10 V495_K118Q_S177E YES
12 G07 V495_Y216A_V294N YES
14 C06 Q161A_M162F_Q295A YES
15 H08 M106E_M162A_Y216A YES
16 A04 L219F_V294N_Q295A YES
18 C08 V495_S214G_V294A YES
20 H10 M162A_N173D_S214F YES
22 G05 A53T_K118N_S214F YES
23 F09 Q38G_D166E_Q295A YES
24 D11 F123H_L174V_S177E YES
For the singleton and doubleton mutants resulting from the breakdown of triple mutants—A09; H02; C05; E09; D04; H11; A04; G05; F09; and D11—the total amount of prenylated products (and the respective proportion of CBGVA and 5-GDVA); and % CBGVA within the prenylated products was calculated.
For the remaining triple mutants in Table 6, the breakdown of the triple mutants into singletons and doubletons; and the determination of the total amount of prenylated products and % CBGVA produced by these mutants will be determined.
FIGS. 30-39 depict the total amount of prenylated products and % CBGVA produced using DVA as substrate and GPP as donor for the mutants derived from A09 (FIG. 30); H02 (FIG. 31); C05 (FIG. 32); E09 (FIG. 33); D04 (FIG. 34); H11 (FIG. 35); A04 (FIG. 36); G05 (FIG. 37); F09 (FIG. 38); and D11 (FIG. 39). The % CBGVA for these clones, along with the mutations they carry, are listed in Table 13.
TABLE 13
RBP
CLONE %
ID Mutations CBGVA
A09 V49A_Q161S_V294A 84.59%
022 V49A_Q161S 85.83%
023 V49A_V294A 71.18%
024 Q161S_V294A 47.59%
041 Q161S 51.78%
049 V294A 16.99%
051 V49A 64.92%
H02 A53Q_S177W_L219F 60.64%
007.1 A53Q_S177W 62.28%
008 A53Q_L219F 11.13%
009 S177W_L219F 63.48%
032 A53Q 11.45%
039.2 L219F 17.68%
046 S177W 74.01%
C05 A53Q_S177Y_Y288H 85.70%
019 A53Q_S177Y 17.32%
020 A53Q_Y288H 83.61%
021 S177Y_Y288H 88.73%
032 A53Q 11.45%
047.2 S177Y 25.03%
052 Y288H 86.99%
E09 A53T_M106E_Q161S 29.97%
025 A53T_M106E 15.99%
026 A53T_Q161S 32.84%
027 M106E_Q161S 39.28%
033 A53T 8.75%
040 M106E 29.24%
041 Q161S 51.78%
D04 A53T_D166E_Q295W 43.98%
016 A53T_D166E 5.02%
017 A53T_Q295W 70.34%
018 D166E_Q295W 63.59%
033 A53T 8.75%
034 D166E 13.49%
043 Q295W 78.32%
H11 A108G_Q161S_G205M 47.09%
010 A108G_Q161S 42.42%
011 A108G_G205M 17.79%
012 Q161S_G205M 38.12%
031 A108G 18.18%
036 G205M 14.04%
041 Q161S 51.78%
A04 L219F_V294N_Q295A 16.74%
004 L219F_V294N 13.81%
005.1 L219F_Q295A 20.37%
006 V294N_Q295A 14.19%
039.2 L219F 17.68%
042 Q295A 24.51%
050 V294N 14.42%
G05 A53T_K118N_S214F 30.01%
028 A53T_K118N 6.24%
029.1 A53T_S214F 31.46%
030 K118N_S214F 44.77%
033 A53T 8.75%
037 K118N 17.50%
048 S214F 51.33%
F09 Q38G_D166E_Q295A 37.37%
001 Q38G_D166E 14.60%
002 Q38G_Q295A 18.61%
003 D166E_Q295A 17.65%
034 D166E 13.49%
042 Q295A 24.51%
044 Q38G 17.66%
D11 F123H_L174V_S177E 33.76%
013 F123H_L174V 14.04%
014 F123H_S177E 32.66%
015 L174V_S177E 86.04%
035 F123H 12.63%
045 S177E 88.33%
038 L174V 19.90%
G12 A17T_Q161W_A232S 89.66%
q423-1 A17T 13.71%
EE09 Q161W 4.21%
424-1 A232S 80.92%
C06 Q161A_M162F_Q295A 49.09%
DD01 Q161A 26.80%
431-1 M162F 39.68%
042.3 Q295A 23.80%
F08 A53T_N173D_S214R 61.02%
033 A53T 8.75%
427-4 N173D 16.18%
BB05 S214R 100.00%
This analysis provided important insights into which positions on ORF2, when mutated, are likely to give rise to significant effects on the enzymatic activity of ORF2 in the reaction using DVA as substrate and GPP as donor. Based on this analysis, the following amino acid sites were selected for targeted amino acid site saturation mutagenesis—V49, Q161, 5177, Y288, M106, Q295, 5214, M162 and A232.
Site saturated mutants of Q295, Q161, 5214 and Y288, described in Example 1, were analyzed to determine the amount of total prenylated products they produce; their respective CBGVA production potential; and 5-GDVA production potential using DVA as substrate and GPP as donor. These results are depicted in FIG. 40 (Q295), FIG. 41 (Q161), FIG. 42 (S214) and FIG. 98 (Y288); and in Tables 14-17.
TABLE 14
Q295 site saturated mutants
CBGVA 5-GDVA
nMol nMol 5- Total % % Production % 5- Production
Mutations CBGVA GDVA Products CBGVA Activity Potential GDVA Potential
Q295D 1.29140226 9.345758355 10.63716061 12.14% 155.20% 0.19 87.86% 1.36
Q295Y 1.501276892 6.363975677 7.865252569 19.09% 114.76% 0.22 80.91% 0.93
Q295N 0.511492029 5.062710105 5.574202134 9.18% 60.35% 0.06 90.82% 0.55
Q295H 3.540705773 4.739840815 8.280546588 42.76% 89.65% 0.38 57.24% 0.51
Q295G 0.703722334 4.449401819 5.153124153 13.66% 55.79% 0.08 86.34% 0.48
Q295S 0.480420987 3.918108562 4.39852955 10.92% 47.62% 0.05 89.08% 0.42
Q295K 0.249922613 2.476591853 2.726514465 9.17% 39.78% 0.04 90.83% 0.36
Q295A 0.775344374 2.481832114 3.257176488 23.80% 35.27% 0.08 76.20% 0.27
Q295C 1.126799257 2.37638422 3.503183477 32.17% 37.93% 0.12 67.83% 0.26
Q295R 0.038925863 1.778425944 1.817351807 2.14% 19.68% 0.00 97.86% 0.19
Q295W 4.568410463 1.598897568 6.167308031 74.07% 66.77% 0.49 25.93% 0.17
Q295M 3.386047052 1.594695472 4.980742523 67.98% 53.93% 0.37 32.02% 0.17
Q295T 0.343174431 1.441393118 1.78456755 19.23% 19.32% 0.04 80.77% 0.16
Q295L 2.095728215 0.998467471 3.094195686 67.73% 33.50% 0.23 32.27% 0.11
Q295P 0.579322086 0.893439786 1.472761873 39.34% 15.95% 0.06 60.66% 0.10
Q295E 0.68298251 0.480324303 1.163306813 58.71% 12.60% 0.07 41.29% 0.05
Q295F 13.57351803 0.283839233 13.85735726 97.95% 150.03% 1.47 2.05% 0.03
Q295V 0.736921529 0.261963615 0.998885144 73.77% 10.81% 0.08 26.23% 0.03
Q295I 3.401176289 0.248071979 3.649248268 93.20% 39.51% 0.37 6.80% 0.03
TABLE 15
Q161 site saturated mutants
CBGVA 5-GDVA
nMol nMol 5- Total % % Production % 5- Production
Mutations CBGVA GDVA Products CBGVA Activity potential GDVA Potential
Q161C 0.342207089 10.68046767 11.02267476 3.10% 185.25% 0.06 96.90% 1.79495767
Q161M 0.526466491 10.36728792 10.89375441 4.83% 183.08% 0.09 95.17% 1.742324731
Q161T 2.700278595 7.140523037 9.840801632 27.44% 165.38% 0.45 72.56% 1.200035147
Q161L 0.836673889 6.451057939 7.287731829 11.48% 122.48% 0.14 88.52% 1.084163754
Q161F 3.890728989 6.157108958 10.04783795 38.72% 168.86% 0.65 61.28% 1.034762736
Q161I 0.501315586 6.070026696 6.571342281 7.63% 110.44% 0.08 92.37% 1.020127705
Q161S 4.812335552 4.916106387 9.728441939 49.47% 163.50% 0.81 50.53% 0.826200045
Q161N 0.992802972 4.474688551 5.467491522 18.16% 79.77% 0.14 81.84% 0.652878827
Q161A 1.775886086 4.850479533 6.626365619 26.80% 77.10% 0.21 73.20% 0.56439113
Q161W 0.193778053 4.411879573 4.605657626 4.21% 52.93% 0.02 95.79% 0.507033637
Q161V 0.594335242 3.318741349 3.913076591 15.19% 47.18% 0.07 84.81% 0.400147091
Q161G 0.999961306 2.683112517 3.683073824 27.15% 53.74% 0.15 72.85% 0.391479168
Q161P 0.78366352 2.170333202 2.953996721 26.53% 33.95% 0.09 73.47% 0.249424745
Q161R 9.435304132 0.931753016 10.36705715 91.01% 174.23% 1.59 8.99% 0.156590261
Q161E 1.672070887 1.24184299 2.913913877 57.38% 35.13% 0.20 42.62% 0.149731422
Q161D 1.211654543 1.262013051 2.473667594 48.98% 28.43% 0.14 51.02% 0.145036386
Q161K 5.035404736 0.710945224 5.746349961 87.63% 66.86% 0.59 12.37% 0.082724023
Q161Y 8.142121963 0.489148705 8.631270667 94.33% 145.06% 1.37 5.67% 0.082206252
Q161H 31.43685188 0 31.43685188 100.00% 365.79% 3.66 0.00% 0
TABLE 16
S214 site saturated mutants
CBGVA 5-GDVA
nMol nMol 5- Total % % Production % 5- Production
Mutations CBGVA GDVA Products CBGVA Activity Potential GDVA Potential
S214T 1.3722334 5.235391536 6.607624937 20.77% 0.964084621 0.200215528 79.23% 0.763869093
S214C 0.265206624 5.436770813 5.701977437 4.65% 0.758613666 0.03528414 95.35% 0.723329526
S214A 0.24686581 5.326280403 5.573146214 4.43% 0.671055324 0.029724793 95.57% 0.641330531
S214V 0.854859929 3.382539055 4.237398984 20.17% 0.618257125 0.124728222 79.83% 0.493528902
S214G 1.124864572 3.627545976 4.752410548 23.67% 0.561118545 0.132813099 76.33% 0.428305446
S214N 1.656013001 1.869166502 3.525179503 46.98% 0.514340838 0.241620353 53.02% 0.272720485
S214D 1.865500696 1.657010085 3.522510782 52.96% 0.513951458 0.272185626 47.04% 0.241765832
S214E 1.805254605 0.375568519 2.180823123 82.78% 0.318192702 0.263395428 17.22% 0.054797274
S214F 0.415609039 0.364890251 0.78049929 53.25% 0.113878643 0.060639381 46.75% 0.053239263
S214Q 1.803900325 0.430838442 2.234738767 80.72% 0.269082003 0.217205304 19.28% 0.051876699
S214I 0.482239591 0.348279612 0.830519204 58.06% 0.121176792 0.070361102 41.94% 0.05081569
S214P 0.015206624 0.291032233 0.306238857 4.97% 0.040743231 0.00202315 95.03% 0.038720081
S214K 1.887556106 0.314489816 2.202045922 85.72% 0.25622522 0.219631877 14.28% 0.036593343
S214M 0.227093329 0.106115286 0.333208615 68.15% 0.048616758 0.033134021 31.85% 0.015482736
S214W 0.016483516 0.099070595 0.115554112 14.26% 0.016859907 0.002405025 85.74% 0.014454882
S214Y 0.077619564 0.052995847 0.130615411 59.43% 0.019057424 0.011325072 40.57% 0.007732352
S214L 0.054171181 0.021628436 0.075799617 71.47% 0.011059533 0.007903839 28.53% 0.003155694
S214R 13.19528711 0 13.19528711 100.00% 1.925256578 1.925256578 0.00% 0
S214H 10.22125058 0 10.22125058 100.00% 1.491330181 1.491330181 0.00% 0
TABLE 17
Y288 site saturated mutants
nMol nMol 5- Total % % CBGVA % 5- 5-GDVA
Mutations CBGVA GDVA Products CBGVA Activity Production GDVA Prod
Y288F 33.52313883 0.145837453 33.66897629 99.57% 405.95% 4.04 0.43% 0.02
Y288I 25.12084043 0.138421989 25.25926242 99.45% 304.56% 3.03 0.55% 0.02
Y288L 22.54945055 0.325439984 22.87489053 98.58% 275.81% 2.72 1.42% 0.04
Y288M 21.39599133 0.146035199 21.54202653 99.32% 259.74% 2.58 0.68% 0.02
Y288P 12.60462777 0.052674511 12.65730228 99.58% 152.61% 1.52 0.42% 0.01
Y288T 9.939676521 0.141487048 10.08116357 98.60% 121.55% 1.20 1.40% 0.02
Y288W 8.629972141 0.152264188 8.782236329 98.27% 105.89% 1.04 1.73% 0.02
Y288C 6.701865036 0 6.701865036 100.00% 80.81% 0.81 0.00% 0.00
Y288A 5.34870763 0 5.34870763 100.00% 64.49% 0.64 0.00% 0.00
Y288G 5.26087293 0.108587107 5.369460037 97.98% 64.74% 0.63 2.02% 0.01
Y288H 1.234058195 0.204419616 1.438477812 85.79% 17.34% 0.15 14.21% 0.02
Y288S 1.190102151 0.026522642 1.216624793 97.82% 14.67% 0.14 2.18% 0.00
Y288N 0.893437548 0.060806802 0.954244351 93.63% 11.49% 0.11 6.37% 0.01
Y288K 0.856485064 0.547137631 1.403622695 61.02% 16.92% 0.10 38.98% 0.07
Y288D 0.358651911 0.01977457 0.378426481 94.77% 4.56% 0.04 5.23% 0.00
Y288R 0.156786875 0.012878189 0.169665064 92.41% 2.05% 0.02 7.59% 0.00
Y288E 0.139684259 0.017401622 0.157085881 88.92% 1.89% 0.02 11.08% 0.00
In a similar manner, site saturated mutants of V49, 5177, and M106 will also be generated. Mutant ORF2 enzymatic function using DVA as substrate and GPP as donor for each of these site saturated mutants will be analyzed as described above.
From the results described above, multiple mutations of Q295, Q161, Y288 and 5214 that have significantly higher CBGVA production potential and/or the total amount of prenylated products, as compared to WT ORF2, were identified. Thus, the ORF2 mutants disclosed herein have unexpectedly superior enzymatic functions, in a reaction using DVA as a substrate and GPP as donor, as compared to WT ORF2.
Finally, ORF2 stacking mutants that carry different novel combinations of the mutations that our analysis identified as being important for ORF2's enzymatic activity were analyzed to obtain the total amount of prenylated products produced by each of the stacking mutants (FIG. 43); % enzymatic activity (FIG. 44), % CBGVA produced (FIG. 45), and CBGVA production potential (FIG. 46). Table 18 provides a summary of the enzymatic function, using DVA as substrate and GPP as donor, for each of the stacking mutants.
The analysis of the stacking mutants shows that multiple stacking mutants have significantly higher % enzymatic activity, % CBGVA, and CBGVA production potential, compared to the WT ORF2, thereby indicating that the ORF2 stacking mutants disclosed herein have synergistically enhanced effects compared to the individual single mutants. Thus, the ORF2 stacking mutants disclosed herein have unexpectedly superior enzymatic functions, in a reaction using DVA and GPP, as compared to WT ORF2.
For instance, ORF2 double mutants, A53T-S214R; Q161S-Q295L; Q161S-Q295F; S177W-Q295A; A53T-Q295F; and Q161S-S177W have synergistically enhanced CBGVA production potential and % activity as compared to either of the single mutants. See FIGS. 86-91.
More stacking mutants will be generated as described above, based on the planned breakdown analysis of triple mutants described above, and planned site saturation mutagenesis experiments described above. These stacking mutants will further be analyzed to determine their % enzymatic activity, % CBGVA, %5-GDVA and CBGVA production potential.
TABLE 18
CBGVA 5-GDVA nMol nMol 5- Total % % CBGVA
Mutations (6.39) (6.458) CBGVA GDVA Products CBGVA Activity Production
A53T_S214R 63.8414 24.70260022 0 24.70260022 100.00% 287.43% 2.9
Q161S_Q295L 54.3225 1.5682 21.01938554 0.387631007 21.40701655 98.19% 249.09% 2.4
S177W_Q295A 49.6105 19.19613837 0 19.19613837 100.00% 223.36% 2.2
Q161S_Q295F 46.2151 1.8605 17.88233246 0.459882341 18.3422148 97.49% 213.43% 2.1
A53T_Q295F 43.9523 2.8376 17.0067714 0.701403994 17.70817539 96.04% 206.05% 2.0
S214R_Q295F 35.9686 13.91758242 0 13.91758242 100.00% 161.94% 1.6
Q161S_S177W 32.4331 2.5717 12.54956663 0.6350678268 13.1852449 95.18% 153.42% 1.5
Q16S_S214R 29.2403 11.31415416 0 11.31415416 100.00% 131.65% 1.3
A53T_S177W 15.4625 8.8346 5.983013465 2.183755191 8.166768656 73.26% 95.03% 0.7
A53T_V294A_Q295W 14.5967 8.5727 5.648003405 2.119018193 7.767021598 72.72% 84.09% 0.6
A53T_Q161S_V294A_Q295W 13.4626 11.7614 5.209178146 2.907207831 8.116385977 64.18% 87.88% 0.6
A53T_Q295W 12.1532 7.1918 4.702522829 1.777684398 6.480207227 72.57% 70.16% 0.5
Q161S_V294A_Q295A 11.5893 6.1801 4.484329051 1.527610243 6.011939294 74.59% 65.09% 0.5
A53T_Q161S_V294N_Q295W 11.3653 13.5299 4.397655162 3.344349417 7.742004578 56.80% 83.82% 0.5
A53T_Q161S 10.4118 26.798 4.028710726 6.623986553 10.65269728 37.82% 123.95% 0.5
A53T_Q161S_Q295W 10.4226 5.6942 4.032889646 1.407504449 5.440394095 74.13% 58.90% 0.4
S177W_S214R 8.898 3.442965485 0 3.442965485 100.00% 40.06% 0.4
A53T_Q161S_V294A_Q295A 8.3752 9.5435 3.240674818 2.358982598 5.599657417 57.87% 60.63% 0.4
A53T_Q161S_V294N_Q295A 7.7662 7.1111 3.005030181 1.7577368 4.762766982 63.09% 51.57% 0.3
A53T_Q161S_Q295A 6.9677 7.6228 2.696060981 1.884219893 4.580280874 58.86% 49.59% 0.3
A53T_Q161S_V294A 5.7154 18.9083 2.211499768 4.673793751 6.885293519 32.12% 74.55% 0.2
Q161S_V294A_Q295W 5.3676 3.451 2.076923077 0.853025509 2.929948586 70.89% 31.72% 0.2
A53T_Q161S_V294N 4.6934 17.5207 1.816050147 4.330803836 6.146853983 29.54% 66.55% 0.2
V49A_S214R 3.7232 0.2083 1.440643863 0.051488036 1.4921319 96.55% 17.36% 0.2
A53T_V294A 2.2627 34.0135 0.875522365 8.407529167 9.283051532 9.43% 100.51% 0.1
A53T_V294A_Q295A 1.874 10.6083 0.72511995 2.622182124 3.347302074 21.66% 36.24% 0.1
A53T_Q295A 1.7119 12.918 0.662397462 3.193098675 3.855496137 17.18% 41.74% 0.1
V49A_Q295L 1.4388 1.2094 0.556724965 0.298942061 0.855667026 65.06% 9.96% 0.1
Example 5: Generation of ORF2 Variants with Improved Enzymatic Function Using DVA as Substrate and GPP as Donor Four triple mutants (A09 carrying V49A_Q161S_V294A; E9 carrying A53T_M106E_Q161S; G12 carrying A17T_Q161W_A232S; and D12 carrying A53T_E112D_G205M) that had improved activity vs. the WT ORF2, based on FIG. 27 were targeted for “breakdown” analysis. For each parental clone targeted six unique mutants are generated (3 doubles and 3 singles).
The % enzymatic activity when using DVA as substrate and GPP as donor was calculated for each of the single and double mutants derived from the breakdown analysis of A09 and E09. FIGS. 47 and 48 depict the % activity for the mutants derived from the breakdown of A09 (FIG. 47); and E09 (FIG. 48) triple mutant clones.
In a similar fashion, breakdown analysis for the other two triple mutants—G12 and D12—will also be done; and the % activity for the mutants derived from the breakdown will be measured, as described above.
Based on the breakdown analysis of triple mutants A09 and E09: V49, Q161, A53 and Q295 were selected for performing site-saturated mutagenesis. % enzymatic activity (when using DVA as substrate and GPP as donor) of site-saturated mutants of A53, Q295 and Q161 described in Examples 1 and 2 was determined, and was compared to that of WT ORF2. These results are shown in FIG. 49 (A53), FIG. 50 (Q295), and FIG. 51 (Q161); and Tables 19, 20 and 21.
TABLE 19
Site saturated mutants of A53
nMol nMol Total %
Mutations CBGVA 5-GDVA Products Activity
A53L 2.143514936 9.210351987 11.35386692 190.81%
A53K 1.275383068 8.751705557 10.02708862 168.52%
A53V 2.534553475 7.935633775 10.47018725 152.77%
A53M 1.582688438 8.177328456 9.760016894 142.40%
A53Q 1.078548212 7.218162942 8.296711155 139.43%
A53T 1.075065779 10.81335278 11.88841856 136.63%
A53S 1.338492493 7.507118845 8.845611339 129.06%
A53C 1.754449776 7.060955112 8.815404887 1.06289
A53I 2.838724656 6.020367807 8.859092463 101.81%
A53N 0.540164061 4.653129326 5.193293387 75.77%
A53R 0.705811794 5.140523037 5.846334831 67.19%
A53W 0.3875948 5.026596797 5.414191596 62.22%
A53G 0.675475933 3.400830532 4.076306464 59.48%
A53D 0.480691843 2.19952541 2.680217254 45.04%
A53Y 0.493151215 1.538090765 2.03124198 34.14%
A53F 0.470863643 0.246860787 0.71772443 8.35%
A53H 0.167311562 0.30047459 0.467786151 5.44%
A53E 0.032154465 0.156837058 0.188991523 3.18%
A53P 0.000580406 0.021925054 0.02250546 0.002714
TABLE 20
Site saturated mutants of Q295
CBGVA
nMol nMol 5- Total % % Production
Mutations CBGVA GDVA Products CBGVA Activity Potential
Q295D 1.29140226 9.345758355 10.63716061 12.14% 155.20% 0.19
Q295F 13.57351803 0.283839233 13.85735726 97.95% 150.03% 1.47
Q295Y 1.501276892 6.363975677 7.865252569 19.09% 114.76% 0.22
Q295H 3.540705773 4.739840815 8.280546588 42.76% 89.65% 0.38
Q295W 4.568410463 1.598897568 6.167308031 74.07% 66.77% 0.49
Q295N 0.511492029 5.062710105 5.574202134 9.18% 60.35% 0.06
Q295G 0.703722334 4.449401819 5.153124153 13.66% 55.79% 0.08
Q295M 3.386047052 1.594695472 4.980742523 67.98% 53.93% 0.37
Q295S 0.480420987 3.918108562 4.39852955 10.92% 47.62% 0.05
Q295K 0.249922613 2.476591853 2.726514465 9.17% 39.78% 0.04
Q295I 3.401176289 0.248071979 3.649248268 93.20% 39.51% 0.37
Q295C 1.126799257 2.37638422 3.503183477 32.17% 37.93% 0.12
Q295A 0.775344374 2.481832114 3.257176488 23.80% 35.27% 0.08
Q295L 2.095728215 0.998467471 3.094195686 67.73% 33.50% 0.23
Q295R 0.038925863 1.778425944 1.817351807 2.14% 19.68% 0.00
Q295T 0.343174431 1.441393118 1.78456755 19.23% 19.32% 0.04
Q295P 0.579322086 0.893439786 1.472761873 39.34% 15.95% 0.06
Q295E 0.68298251 0.480324303 1.163306813 58.71% 12.60% 0.07
Q295V 0.736921529 0.261963615 0.998885144 73.77% 10.81% 0.08
TABLE 21
Site saturated mutants of Q161
nMol nMol 5- Total % % CBGVA
Mutations CBGVA GDVA Products CBGVA Activity Production
Q161H 31.43685188 0 31.43685188 100.00% 365.79% 3.66
Q161C 0.342207089 10.68046767 11.02267476 3.10% 185.25% 0.06
Q161M 0.526466491 10.36728792 10.89375441 4.83% 183.08% 0.09
Q161R 9.435304132 0.931753016 10.36705715 91.01% 174.23% 1.59
Q161F 3.890728989 6.157108958 10.04783795 38.72% 168.86% 0.65
Q161T 2.700278595 7.140523037 9.840801632 27.44% 165.38% 0.45
Q161S 4.812335552 4.916106387 9.728441939 49.47% 163.50% 0.81
Q161Y 8.142121963 0.489148705 8.631270667 94.33% 145.06% 1.37
Q161L 0.836673889 6.451057939 7.287731829 11.48% 122.48% 0.14
Q161I 0.501315586 6.070026696 6.571342281 7.63% 110.44% 0.08
Q161N 0.992802972 4.474688551 5.467491522 18.16% 79.77% 0.14
Q161A 1.775886086 4.850479533 6.626365619 26.80% 77.10% 0.21
Q161K 5.035404736 0.710945224 5.746349961 87.63% 66.86% 0.59
Q161G 0.999961306 2.683112517 3.683073824 27.15% 53.74% 0.15
Q161W 0.193778053 4.411879573 4.605657626 4.21% 52.93% 0.02
Q161V 0.594335242 3.318741349 3.913076591 15.19% 0.471807246 0.07
Q161E 1.672070887 1.24184299 2.913913877 57.38% 0.351336257 0.20
Q161P 0.78366352 2.170333202 2.953996721 26.53% 33.95% 0.09
Q161D 1.211654543 1.262013051 2.473667594 48.98% 28.43% 0.14
Similarly, site saturated mutants of V49 will also be generated; and the % activity of these mutants will be compared to that of the WT ORF2 in a reaction using DVA as substrate and GPP as donor.
From the results described above, multiple mutations of A53, Q295, and Q161 that have significantly higher % activity, as compared to WT ORF2, were identified. Thus, the ORF2 mutants disclosed herein have unexpectedly superior enzymatic functions, in a reaction using DVA as a substrate and GPP as donor, as compared to WT ORF2.
Finally, ORF2 stacking mutant clones that carry different novel combinations of the mutations that our analysis identified as being important for ORF2's % enzymatic activity were analyzed to measure their % enzymatic activity, as shown in FIG. 52.
The analysis of the stacking mutants shows that multiple stacking mutants have significantly higher % enzymatic activity, compared to the WT ORF2, thereby indicating that the ORF2 stacking mutants disclosed herein have synergistically enhanced effects compared to the individual single mutants. Thus, the ORF2 stacking mutants disclosed herein have unexpectedly superior enzymatic functions, in a reaction using DVA and GPP, as compared to WT ORF2.
Example 6: Generation of ORF2 Variants which Synthesize an Altered Amount of CBG and/or 5-GO Using Olivetol (Referred to Herein as “O”) and GPP, Compared to WT ORF2 The library of tripleton mutants was screened for the ability to produce CBG and 5-GO using O as substrate and Geranyl pyrophosphate (GPP) as donor; and this enzymatic activity was compared to that of the WT ORF2. Table 22 provides the summary of the analysis performed on the enzymatic activity of the ORF2 triple mutants to produce CBG and 5-GO using O as substrate and Geranyl pyrophosphate (GPP) as donor. Table 9 lists the mutations within each of the tripleton mutants as well the nMol CBG, nMol of 5-GO, total prenylated products, % CBG products among prenylated products, % enzymatic activity, CBG production potential and %5-GO among total prenylated products of each of the ORF2 triple mutants.
TABLE 22
CLONE CBG 5-GO Total nMol % CBG
ID Mutations (7.095) (7.745) nMol CBG nMol 5-GO Products % CBG Activity Production % GOA
E09 A53T_M106E_Q161S 3.1916 1.3656 13.04290969 3.762190754 16.80510044 77.61% 406.28% 3.15 22.39%
C06 Q161A_M162F_Q295A 1.8681 0.787 7.634246016 2.168163535 9.802409551 77.88% 219.65% 1.71 22.12%
D12 A53T_E112D_G205M 1.5896 1.2489 6.496117695 3.440685437 9.936803132 65.37% 240.23% 1.57 34.63%
G12 A17T_Q161W_A232S 1.0678 0.4634 4.363710666 1.276654361 5.640365027 77.37% 136.36% 1.05 22.63%
C11 E112D_L219F_V294F 0.8983 0.8995 3.671025746 2.478097967 6.149123713 59.70% 137.79% 0.82 40.30%
A09 V49A_Q161S_V294A 0.703 0.8977 2.872905599 2.473139016 5.346044615 53.74% 119.79% 0.64 46.26%
D06 A53E_Q161A_V294N 0.5894 0.7298 2.40866367 2.010579095 4.419242765 54.50% 99.03% 0.54 45.50%
A04 L219F_V294N_Q295A 0.5735 0.4173 2.343686146 1.149650118 3.493336265 67.09% 78.28% 0.53 32.91%
H11 A108G_Q161S_G205M 0.4996 0.4828 2.041683694 1.330100832 3.371784526 60.55% 81.52% 0.49 39.45%
H02 A53Q_S177W_L219F 0.4683 0.4481 1.913771966 1.234503278 3.148275244 60.79% 76.11% 0.46 39.21%
D04 A53T_D166E_Q295W 0.405 0.6084 1.655087863 1.676125406 3.331213269 49.68% 74.65% 0.37 50.32%
H09 E112G_G205M_L298W 0.355 0.6902 1.450756028 1.901482175 3.352238203 43.28% 81.04% 0.35 56.72%
C05 A53Q_S177Y_Y288H 0.371 0.703 1.516142215 1.936745826 3.452888041 43.91% 77.37% 0.34 56.09%
F09 Q38G_D166E_Q295A 0.3134 0.3094 1.280751941 0.852388561 2.133140503 60.04% 51.57% 0.31 39.96%
H10 M162A_N173D_S214F 0.3072 0.5322 1.255414794 1.466196485 2.721611278 46.13% 65.80% 0.30 53.87%
H06 V49L_E112D_G286E 0.3004 0.3258 1.227625664 0.897570114 2.125195778 57.77% 51.38% 0.30 42.23%
A05 A17T_C25V_E112G 0.3182 0.3457 1.300367797 0.952394071 2.252761869 57.72% 50.48% 0.29 42.28%
D11 F123H_L174V_S177E 0.2886 0.3135 1.179403351 0.86368395 2.043087301 57.73% 49.39% 0.29 42.27%
B08 K118Q_L174V_R228Q 0.3098 0.3717 1.266040049 1.024023362 2.290063411 55.28% 51.32% 0.28 44.72%
A02 Q38G_E112D_F123H 0.3065 0.4033 1.252554148 1.1110805 2.363634648 52.99% 52.96% 0.28 47.01%
G05 A53T_K118N_S214F 0.2765 0.3188 1.129955047 0.878285305 2.008240352 56.27% 48.55% 0.27 43.73%
B10 M106E_Y121W_D166E 0.2878 0.3451 1.176134042 0.950741088 2.126875129 55.30% 47.66% 0.26 44.70%
H05 S177E_S214R_R228E 0.2652 0.3543 1.083776052 0.976086837 2.059862889 52.61% 49.80% 0.26 47.39%
C12 N173D_F213M_V294F 0.2854 0.6328 1.166326114 1.743346741 2.909672854 40.08% 65.20% 0.26 59.92%
B09 C25V_F213M_Y216A 0.2775 0.3398 1.134041684 0.936139732 2.070181416 54.78% 46.39% 0.25 45.22%
D01 K118Q_Q161W_S214F 0.2761 0.3493 1.128320392 0.962311973 2.090632365 53.97% 46.85% 0.25 46.03%
C02 K118N_ K119A_V271E 0.2744 0.3159 1.12137311 0.870295884 1.991668994 56.30% 44.63% 0.25 43.70%
B06 D166E_S177Y_S214F 0.2696 0.3306 1.101757254 0.910793983 2.012551237 54.74% 45.10% 0.25 45.26%
B05 A53Q_Y121W_A232S 0.2661 0.3326 1.087454025 0.916303929 2.003757954 54.27% 44.90% 0.24 45.73%
G03 L219F_Y283L_L298W 0.2448 0.4232 1.000408664 1.165904458 2.166313121 46.18% 52.37% 0.24 53.82%
H01 K119A_Q161A_R228Q 0.24 0.3273 0.980792808 0.901702573 1.882495381 52.10% 45.51% 0.24 47.90%
G09 M106E_G205L_C209G 0.2391 0.323 0.977114834 0.88985619 1.866971025 52.34% 45.14% 0.24 47.66%
C03 V49L_S214R_V271E 0.2545 0.3185 1.04004904 0.877458813 1.917507853 54.24% 42.97% 0.23 45.76%
G06 K118Q_F123A_R228E 0.2329 0.3136 0.951777687 0.863959447 1.815737134 52.42% 43.90% 0.23 47.58%
A12 Y121W_S177Y_G286E 0.2499 0.3016 1.021250511 0.830899774 1.852150285 55.14% 41.50% 0.23 44.86%
E10 E112D_K119A_N173D 0.2314 0.2803 0.945647732 0.772218855 1.717866587 55.05% 41.53% 0.23 44.95%
G11 S177W_Y288H_V294N 0.2266 0.3002 0.926031876 0.827042812 1.753074688 52.82% 42.38% 0.22 47.18%
B04 A53E_A108G_K118N 0.2397 0.2981 0.979566817 0.82125737 1.800824186 54.40% 40.35% 0.22 45.60%
G10 V49A_Y121W_C230S 0.2207 0.299 0.901920719 0.823736845 1.725657564 52.27% 41.72% 0.22 47.73%
F08 A53T_N173D_S214R 0.2207 0.2818 0.901920719 0.776351314 1.678272033 53.74% 40.57% 0.22 46.26%
G07 V49S_Y216A_V294N 0.2206 0.3124 0.901512056 0.86065348 1.762165535 51.16% 42.60% 0.22 48.84%
H07 F123A_M162F_S214G 0.2201 0.3228 0.899468737 0.889305196 1.788773933 50.28% 43.25% 0.22 49.72%
C07 V49L_K119D_G205M 0.2333 0.3044 0.953412342 0.838613698 1.792026039 53.20% 40.16% 0.21 46.80%
F12 A17T_V49A_C230N 0.2159 0.2812 0.882304863 0.77469833 1.657003194 53.25% 40.06% 0.21 46.75%
D03 D227E_C230N_Q295W 0.2291 0.2973 0.936248468 0.819053391 1.755301859 53.34% 39.33% 0.21 46.66%
C10 A53Q_L274V_Q295A 0.2286 0.298 0.934205149 0.820981872 1.755187021 53.23% 39.33% 0.21 46.77%
C08 V49S_S214G_V294A 0.2284 0.4829 0.933387822 1.330376329 2.263764151 41.23% 50.73% 0.21 58.77%
G02 A53E_F213M_R228Q 0.2077 0.302 0.848794442 0.832001763 1.680796205 50.50% 40.63% 0.21 49.50%
B11 V49S_K119D_F213M 0.2217 0.2841 0.906007356 0.782687751 1.688695107 53.65% 37.84% 0.20 46.35%
F10 K119D_Q161W_L298Q 0.2054 0.2715 0.839395178 0.747975095 1.587370273 52.88% 38.38% 0.20 47.12%
C01 V49S_M162A_Y283L 0.2213 0.7384 0.904372701 2.034271861 2.938644562 30.78% 65.85% 0.20 69.22%
B02 V49A_S177Y_C209G 0.2202 0.2682 0.899877401 0.738883685 1.638761086 54.91% 36.72% 0.20 45.09%
A11 V49L_D166E_L274V 0.2202 0.2631 0.899877401 0.724833324 1.624710725 55.39% 36.41% 0.20 44.61%
D09 K118N_C209G_R228Q 0.2002 0.2477 0.818144667 0.682406744 1.500551411 54.52% 36.28% 0.20 45.48%
G04 D227E_R228E_L298Q 0.1989 0.2764 0.812832039 0.761474461 1.574306501 51.63% 38.06% 0.20 48.37%
A10 V49S_K118Q_S177E 0.2141 0.2994 0.874948917 0.824838834 1.699787751 51.47% 38.09% 0.20 48.53%
B12 A17T_F123W_L298A 0.2115 0.2931 0.864323662 0.807482506 1.671806168 51.70% 37.46% 0.19 48.30%
C09 A108G_K119D_L298A 0.211 0.2503 0.862280343 0.689569673 1.551850017 55.56% 34.77% 0.19 44.44%
H04 M162A_C209G_Y288H 0.1947 0.2801 0.795668165 0.77166786 1.567336026 50.77% 37.89% 0.19 49.23%
D08 E112D_K119A_N173D 0.1903 0.2403 0.777686964 0.662019946 1.43970691 54.02% 34.81% 0.19 45.98%
A03 V49L_F123A_Y283L 0.1942 0.2095 0.793624847 0.577166786 1.370791633 57.90% 30.72% 0.18 42.10%
A07 G205L_R228E_C230N 0.194 0.2399 0.792807519 0.660917957 1.453725476 54.54% 32.57% 0.18 45.46%
A08 C25V_A232S_V271E 0.1891 0.2273 0.772783 0.626205301 1.3989883 55.24% 31.35% 0.17 44.76%
D07 K119A_S214G_L298A 0.1708 0.2959 0.697997548 0.81519643 1.513193978 46.13% 36.58% 0.17 53.87%
D10 V49A_F123A_Y288H 0.136 0.1827 0.555782591 0.503333517 1.059116108 52.48% 25.61% 0.13 47.52%
The amount of CBG or 5-GO (in nMols) generated by each of the ORF2 triple mutant clones was measured using HPLC. FIG. 53 shows the total nMols of prenylated products generated using O as substrate and GPP as donor by each of the ORF2 mutants, and the proportion of CBG and 5-GO within the total prenylated products. An exemplary wild type ORF2 replicate is included in the graph for comparison purposes.
FIG. 54 shows the % CBG and %5-GO within the total prenylated products produced by each of the ORF2 triple mutant clones using O as substrate and GPP as donor. In this graph, the mutant clones are ordered based on decreasing % CBG (from left to right) they produce, with the % GO depicted in red. The black dashed line on the graph indicates the % CBG that is produced by the wild type enzyme.
FIG. 55 shows the ORF2 enzymatic activity (using O as substrate and GPP as donor) of each of the triple mutant ORF2 clones relative to the wild type enzyme. % activity was calculated by dividing the nMols of total prenylated products produced by a mutant by the nMols of total prenylated products produced by the wild type control, and expressed as a percentage. The red threshold line is the wild type Orf2% activity.
FIG. 56 shows the CBG production potential of each of the ORF2 triple mutant clones when using O as substrate and GPP as donor. CBG production potential (interchangeably referred to herein as CBG production quotient) represents the improvement in CBG production vs. the wild type enzyme. CBG production potential was calculated by multiplying the % CBG of the Total Products by the % Activity of each mutant.
While the CBG production potential analysis shown in FIG. 56 is useful to rank ORF2 mutant clones based on the amount of CBG produced, such an analysis would not differentiate between a mutant that made 100% CBG but was 20% as active as wild type ORF2; or a mutant that made 10% CBG and was 200% as active as wild type ORF2. Therefore, we employed a cluster analysis by plotting the CBG Production Potential vs. %5-GO (FIG. 57). %5-GO was calculated in a similar manner as % CBG. We used the top 25 mutants ranked based on their CBG production potential for this analysis. High 5-GO producing mutants cluster together towards the right of the graph and high CBG producing mutants cluster towards the left of the graph.
Based on the analysis performed in FIG. 57, 19 mutants which cluster to the left of the graph were selected for “breakdown” analysis (Table 23). For each parental clone targeted six unique mutants are generated (3 doubles and 3 singles).
TABLE 23
CBG
Production Clone
Rank ID Mutations
1 E09 A53T_M106E_Q161S
2 C06 Q161A_M162F_Q295A
3 D12 A53T_E112D_G205M
4 G12 A17T_Q161W_A232S
5 C11 E112D_L219F_V294F
6 A09 V49A_Q161S_V294A
7 D06 A53E_Q161A_V294N
8 A04 L219F_V294N_Q295A
9 H11 A108G_Q161S_G205M
10 H02 A53Q_S177W_L219F
11 D04 A53T_D166E_Q295W
13 C05 A53Q_S177Y_Y288H
14 F09 Q38G_D166E_Q295A
16 H06 V49L_E112D_G286E
17 A05 A17T_C25V_E112G
18 D11 F123H_L174V_S177E
19 B08 K118Q_L174V_R228Q
20 A02 Q38G_E112D_F123H
21 G05 A53T_K118N_S214F
For the singleton and doubleton mutants resulting from the breakdown of triple mutants—E09; A09; A04; H11; H02; D04; C05; F09; D011; and G05—the total amount of prenylated products (and the respective proportion of CBG and 5-GO); and % CBG within the prenylated products was calculated. FIGS. 58-67 depict the total amount of prenylated products and % CBG produced using O as substrate and GPP as donor by the mutants derived from E09 (FIG. 58); A09 (FIG. 59); A04 (FIG. 60); H11 (FIG. 61); H02 (FIG. 62); D04 (FIG. 63); C05 (FIG. 64); F09 (FIG. 65); D011 (FIG. 66); and G05 (FIG. 67).
The breakdown analysis of the remaining triple mutants listed in Table 23 will be performed in a similar manner as described above; and the total amount of prenylated products and % CBG produced using O as substrate and GPP as donor by the mutants derived from this breakdown will be measured.
The breakdown analysis of triple mutants—E09; A09; A04; H11; H02; D04; C05; F09; D011; and G05—provided important insights into which positions on ORF2, when mutated, are likely to give rise to significant effects on the enzymatic activity of ORF2 in the reaction using Olivetol (O) as substrate and Geranyl pyrophosphate (GPP) as donor. Based on this analysis, the following amino acid sites were selected for targeted amino acid site saturation mutagenesis—Q161, Q295, 5214, V294 M106, L174, A232, Y288 and M162. Site saturated mutagenesis was done for Q295, Q161, 5214 and Y288 by replacing the wild type residue with each of the other 19 standard amino acids. The amount of total prenylated products produced; CBG production potential; and 5-GO production potential were measured for Q295, Q161, 5214 and Y288 site saturated mutants, as depicted in FIGS. 68-73 and 99; and Tables 25-28.
TABLE 24
RBP
CLONE
ID Mutations % CBG
A09 V49A_Q161S_V294A 53.74%
022 V49A_Q161S 50.81%
023 V49A_V294A 49.55%
024 Q161S_V294A 78.91%
041 Q161S 74.22%
049 V294A 69.60%
051 V49A 50.74%
H02 A53Q_S177W_L219F 60.79%
007.1 A53Q_S177W 66.50%
008 A53Q_L219F 59.04%
009 S177W_L219F 64.31%
032 A53Q 61.21%
039.2 L219F 60.36%
046 S177W 66.96%
C05 A53Q_S177Y_Y288H 43.91%
019 A53Q_S177Y 55.17%
020 A53Q_Y288H 34.48%
021 S177Y_Y288H 50.77%
032 A53Q 61.21%
047.2 S177Y 58.45%
052 Y288H 47.67%
E09 A53T_M106E_Q161S 77.61%
025 A53T_M106E 66.34%
026 A53T_Q161S 77.30%
027 M106E_Q161S 70.34%
033 A53T 68.57%
040 M106E 63.55%
041 Q161S 74.22%
D04 A53T_D166E_Q295W 49.68%
016 A53T_D166E 57.26%
017 A53T_Q295W 61.37%
018 D166E_Q295W 51.62%
033 A53T 68.57%
034 D166E 58.61%
043 Q295W 62.63%
H11 A108G_Q161S_G205M 60.55%
010 A108G_Q161S 62.73%
011 A108G_G205M 56.30%
012 Q161S_G205M 73.18%
031 A108G 54.52%
036 G205M 66.08%
041 Q161S 74.22%
A04 L219F_V294N_Q295A 67.09%
004 L219F_V294N 65.97%
005.1 L219F_Q295A 68.82%
006 V294N_Q295A 71.52%
039.2 L219F 60.36%
042 Q295A 78.68%
050 V294N 69.86%
G05 A53T_K118N_S214F 56.27%
028 A53T_K118N 49.61%
029.1 A53T_S214F 86.94%
030 K118N_S214F 56.78%
033 A53T 68.57%
037 K118N 54.10%
048 S214F 83.66%
F09 Q38G_D166E_Q295A 60.04%
001 Q38G_D166E 56.62%
002 Q38G_Q295A 64.68%
003 D166E_Q295A 67.90%
034 D166E 58.61%
042 Q295A 78.68%
044 Q38G 61.84%
D11 F123H_L174V_S177E 57.73%
013 F123H_L174V 52.69%
014 F123H_S177E 62.24%
015 L174V_S177E 64.87%
035 F123H 52.03%
045 S177E 65.38%
038 L174V 77.09%
G12 A17T_Q161W_A232S 77.37%
q423-1 A17T 68.98%
EE09 Q161W 80.71%
q424-5 A232S 78.59%
C06 Q161A_M162F_Q295A 77.88%
DD01 Q161A 61.26%
431-1 M162F 80.78%
042 Q295A 76.50%
TABLE 25
Q295 site saturated mutants
5-GO
Total nMol CBG % 5- Production
Mutations nMol CBG nMol 5-GO Products % CBG % Activity Production GO Potential
Q295H 26.97874949 8.154443771 35.13319326 76.79% 538.04% 4.13 23.21% 1.25
Q295W 4.693502248 2.936800926 7.630303173 61.51% 116.85% 0.72 38.49% 0.45
Q295F 7.742541888 2.762962147 10.50550403 73.70% 160.88% 1.19 26.30% 0.42
Q295D 2.895790764 1.784120337 4.679911101 61.88% 104.13% 0.64 38.12% 0.40
Q295Y 2.872496935 1.689349275 4.56184621 62.97% 101.50% 0.64 37.03% 0.38
Q295M 9.265631385 2.323819494 11.58945088 79.95% 177.48% 1.42 20.05% 0.36
Q295G 8.638741316 1.972009477 10.61075079 81.41% 162.50% 1.32 18.59% 0.30
Q295C 4.541070699 1.682737341 6.22380804 72.96% 95.31% 0.70 27.04% 0.26
Q295K 1.421332244 1.151578599 2.572910843 55.24% 57.25% 0.32 44.76% 0.26
Q295A 5.303228443 1.62929087 6.932519313 76.50% 106.17% 0.81 23.50% 0.25
Q295E 1.424192889 1.546917186 2.971110075 47.93% 45.50% 0.22 52.07% 0.24
Q295N 2.542705353 1.496225687 4.038931041 62.95% 61.85% 0.39 37.05% 0.23
Q295L 3.336738864 1.470879938 4.807618802 69.41% 73.63% 0.51 30.59% 0.23
Q295S 2.729055987 1.451044135 4.180100122 65.29% 64.02% 0.42 34.71% 0.22
Q295T 2.208418472 1.404209598 3.61262807 61.13% 55.32% 0.34 38.87% 0.22
Q295I 1.582345729 1.128712326 2.711058055 58.37% 41.52% 0.24 41.63% 0.17
Q295V 1.653044544 1.101162598 2.754207143 60.02% 42.18% 0.25 39.98% 0.17
Q295P 1.404168369 0.95652653 2.3606949 59.48% 36.15% 0.22 40.52% 0.15
Q295R 1.035962403 0.816573916 1.852536319 55.92% 28.37% 0.16 44.08% 0.13
TABLE 26
Q161 site saturated mutants
5-GO
Total nMol CBG Production
Mutations nMol CBG nMol 5-GO Products % CBG % Activity Production % 5-GO Potential
Q161L 21.77482632 2.927158521 24.70198484 88.15% 877.78% 7.74 11.85% 1.04
Q161Y 22.34164283 2.522728525 24.86437135 89.85% 883.55% 7.94 10.15% 0.90
Q161F 29.53739273 2.514739104 32.05213183 92.15% 1138.97% 10.50 7.85% 0.89
Q161I 13.8998774 2.498209268 16.39808667 84.77% 582.70% 4.94 15.23% 0.89
Q161C 7.722108705 2.393244807 10.11535351 76.34% 359.45% 2.74 23.66% 0.85
Q161M 14.18716796 1.838944294 16.02611226 88.53% 569.49% 5.04 11.47% 0.65
Q161T 3.827135268 1.707256598 5.534391866 69.15% 196.66% 1.36 30.85% 0.61
Q161H 23.62729873 1.806711114 25.43400985 92.90% 747.62% 6.95 7.10% 0.53
Q161S 3.836534532 1.387679762 5.224214294 73.44% 185.64% 1.36 26.56% 0.49
Q161R 2.120147119 1.386026778 3.506173897 60.47% 124.59% 0.75 39.53% 0.49
Q161G 4.908868002 2.01416056 6.923028561 70.91% 154.03% 1.09 29.09% 0.45
Q161N 3.405803024 1.832883354 5.238686379 65.01% 116.56% 0.76 34.99% 0.41
Q161D 4.249693502 1.342222712 5.591916214 76.00% 164.37% 1.25 24.00% 0.39
Q161A 1.49448304 0.944955645 2.439438685 61.26% 71.71% 0.44 38.74% 0.28
Q161W 3.075194115 0.734751226 3.809945341 80.71% 111.99% 0.90 19.29% 0.22
Q161K 0.852472415 0.651826547 1.504298962 56.67% 44.22% 0.25 43.33% 0.19
Q161P 0.434818145 0.281558213 0.716376357 60.70% 21.06% 0.13 39.30% 0.08
TABLE 27
S214 site saturated mutants
CBG 5-GO
Total nMol Production Production
Mutations nMol CBG nMol 5-GO Products % CBG % Activity potential % 5-GO Potential
S214E 17.95218635 1.994600253 19.9467866 90.00% 443.81% 3.99 10.00% 0.44
S214A 0.523089497 0.539974654 1.063064152 49.21% 0.8036911 0.40 0.507942 0.41
S214I 4.800980793 1.774477933 6.575458725 73.01% 146.30% 1.07 26.99% 0.39
S214T 2.703310176 1.657391592 4.360701768 61.99% 97.02% 0.60 38.01% 0.37
S214F 7.143849612 1.589343765 8.733193377 81.80% 194.31% 1.59 18.20% 0.35
S214M 4.120147119 1.558763568 5.678910687 72.55% 126.35% 0.92 27.45% 0.35
S214L 4.30404577 1.502286627 5.806332398 74.13% 129.19% 0.96 25.87% 0.33
S214G 0.209235799 1.126783845 1.336019644 15.66% 0.3912208 0.06 0.843389 0.33
S214N 7.877400899 1.45352361 9.330924509 84.42% 207.61% 1.75 15.58% 0.32
S214V 2.365753984 1.443054714 3.808808698 62.11% 84.74% 0.53 37.89% 0.32
S214C 0.537801389 0.420684335 0.958485725 56.11% 0.6153359 0.35 0.438905 0.27
S214D 2.427870862 1.186566753 3.614437615 67.17% 80.42% 0.54 32.83% 0.26
S214Y 1.990192072 1.103366577 3.093558649 64.33% 68.83% 0.44 35.67% 0.25
S214H 30.08377605 1.054052565 31.13782862 96.61% 692.81% 6.69 3.39% 0.23
S214W 1.735594606 1.049093614 2.78468822 62.33% 61.96% 0.39 37.67% 0.23
S214R 1.420514916 0.974158356 2.394673272 59.32% 53.28% 0.32 40.68% 0.22
S214Q 37.24560687 0.955975536 38.2015824 97.50% 849.97% 8.29 0.025025 0.21
S214K 1.095218635 0.475508292 1.570726928 69.73% 34.95% 0.24 30.27% 0.11
S214P 0.102983245 0.107719434 0.210702678 48.88% 0.1352685 0.07 0.511239 0.07
TABLE 28
Y288 site saturated mutants
Total nMol CBG % 5- 5-GOA
Mutations nMol CBG nMol 5-GO Products % CBG % Activity Production GOA Production
Y288F 16.59256232 2.590225357 19.18278768 86.50% 561.72% 4.86 13.50% 0.75848
Y288L 13.88516551 1.489338256 15.37450376 90.31% 450.20% 4.07 9.69% 0.43612
Y288I 9.454025337 1.173342884 10.62736822 88.96% 311.20% 2.77 11.04% 0.34358
Y288M 7.780956273 1.176924348 8.957880621 86.86% 262.31% 2.28 13.14% 0.34463
Y288T 5.373518594 0.684059728 6.057578322 88.71% 177.38% 1.57 11.29% 0.20031
Y288C 5.008990601 0.566146895 5.575137496 89.85% 163.25% 1.47 10.15% 0.16578
Y288A 4.105435227 0.304148989 4.409584216 93.10% 129.12% 1.20 6.90% 0.08906
Y288W 2.646505926 0.507741473 3.154247399 83.90% 92.36% 0.77 16.10% 0.14868
Y288P 5.30731508 0.655683509 5.962998588 89.00% 174.61% 0.40 11.00% 0.192
Y288N 0.82958725 0.542729627 1.372316877 60.45% 40.18% 0.24 39.55% 0.15893
Y288S 0.549243972 0.158135434 0.707379407 77.64% 20.71% 0.16 22.36% 0.04631
Y288G 0.398038414 0.08788363 0.485922044 81.91% 14.23% 0.12 18.09% 0.02573
Y288H 0.305271761 0.648245082 0.953516844 32.02% 27.92% 0.09 67.98% 0.18982
Y288K 0.136493666 0.107994931 0.244488597 55.83% 7.16% 0.04 44.17% 0.03162
Y288D 0.097261953 0.073557772 0.170819725 56.94% 5.00% 0.03 43.06% 0.02154
Y288E 0.073968124 0.076312745 0.150280869 49.22% 4.40% 0.02 50.78% 0.02235
Y288R 0.035553739 0.013223869 0.048777608 72.89% 1.43% 0.01 27.11% 0.00387
Similarly, site saturated mutagenesis will also be completed for M106, and V294; and the amount of total prenylated products produced; CBG production potential; and 5-GO production potential will be measured for each of these site saturated mutants.
From the results described above, multiple mutations of Q295, Q161 and 5214 that have significantly higher CBG or 5-GO production potential; and/or higher amount of prenylated products, as compared to WT ORF2, were identified. Thus, the ORF2 mutants disclosed herein have unexpectedly superior enzymatic functions, in a reaction using O as a substrate and GPP as donor, as compared to WT ORF2.
Finally, ORF2 stacking mutants, which carry different novel combinations of the mutations that our analysis identified as being important for ORF2's enzymatic activity were analyzed to determine the total amount of prenylated products produced by each of the stacking mutants (FIG. 74); % enzymatic activity (FIG. 75); % CBG produced (FIG. 76); and CBG production potential (FIG. 77). Table 29 provides a summary of the enzymatic function, using O as substrate and GPP as donor, for each of the stacking mutants. The analysis of the stacking mutants shows that multiple stacking mutants have significantly higher % enzymatic activity, % CBG, and CBG production potential, compared to the WT ORF2, thereby indicating that the ORF2 stacking mutants disclosed herein have synergistically enhanced effects compared to the individual single mutants. Thus, the ORF2 stacking mutants disclosed herein have unexpectedly superior enzymatic functions, in a reaction using O and GPP, as compared to WT ORF2.
For instance, ORF2 double mutants, Q161S-Q295L; A53T-Q161S; A53 T-V294A; Q161S-Q295F; and A53T-Q295F have synergistically enhanced CBGVA production potential and % activity as compared to either of the single mutants. See FIGS. 92-96.
In the future, more stacking mutants will be generated as described above, based on the breakdown analysis of remaining triple mutants listed in Table 10, and planned site saturation mutagenesis experiments described above. These stacking mutants will further be analyzed to determine their % enzymatic activity, % CBG, %5-GO and CBG production potential.
TABLE 29
CBG 5GO Total nMol CBG
Mutations (7.095) (7.745) nMol CBG nMol 5-GO Products % CBG % Activity Production
Q161S_Q295L 3.2538 1.0628 13.29709849 2.927985013 16.2250835 81.95% 628.43% 5.15
A53T_Q161S 3.0213 1.1637 12.34695546 3.205961761 15.55291722 79.39% 602.39% 4.78
A53T_Q161S_V294A_Q295A 6.8655 1.0392 28.05680425 2.862967657 30.91977191 90.74% 473.51% 4.30
Q161S_Q295F 2.6242 0.9004 10.72415202 2.480577442 13.20472947 81.21% 511.44% 4.15
A53T_Q161S_Q295A 6.5383 1.0823 26.71965672 2.981706981 29.7013637 89.96% 454.86% 4.09
A53T_Q295F 2.3515 0.773 9.609726195 2.129593917 11.73932011 81.86% 454.69% 3.72
A53T_Q161S_V294N_Q295A 5.4091 1.0492 22.10502656 2.890517384 24.99554395 88.44% 382.79% 3.39
A53T_Q161S_V294A 5.3751 1.7353 21.96608092 4.780704171 26.74678509 82.13% 409.61% 3.36
Q161S_V294A_Q295A 5.1846 1.1046 21.18757662 3.043142873 24.2307195 87.44% 371.08% 3.24
A53T_Q161S_V294N 4.8641 1.667 19.87780956 4.592539534 24.4703491 81.23% 374.75% 3.04
A53T_V294A 4.3154 2.3259 17.63547201 6.407791063 24.04326307 73.35% 368.21% 2.70
A53T_V294A_Q295A 4.2878 1.2019 17.52268083 3.311201719 20.83388255 84.11% 319.06% 2.68
A53T_Q161S_Q295W 4.2451 1.5899 17.34818145 4.380131137 21.72831258 79.84% 332.75% 2.66
A53T_Q161S_V294N_Q295W 4.1445 1.2834 16.93706579 3.535731996 20.47279779 82.73% 313.53% 2.59
A53T_Q161S_V294A_Q295W 4.1157 1.3136 16.81937066 3.618932173 20.43830283 82.29% 313.00% 2.58
Q161S_V294A_Q295W 2.6247 1.1964 10.72619534 3.296049369 14.02224471 76.49% 214.74% 1.64
A53T_Q295A 2.4689 0.8374 10.08949734 2.307014161 12.3965115 81.39% 189.84% 1.55
A53T_S177W 0.9678 0.4316 3.955046996 1.189046228 5.144093225 76.89% 199.24% 1.53
A53T_V294A_Q295W 2.1217 1.2914 8.670617082 3.557771778 12.22838886 70.91% 187.27% 1.33
A53T_Q295W 2.0002 1.6157 8.174090723 4.451209433 12.62530016 64.74% 193.35% 1.25
S177W_Q295A 0.694 0.4575 2.836125868 1.260400022 4.09652589 69.23% 158.67% 1.10
Q161S_S177W 0.5912 0.4139 2.416019616 1.140283211 3.556302827 67.94% 137.74% 0.94
S214R_Q295F 0.371 0.309 1.516142215 0.851286572 2.367428787 64.04% 91.69% 0.59
A53T_S214R 0.3473 0.2767 1.419288925 0.762300953 2.181589878 65.06% 84.50% 0.55
Q16S_S214R 0.2947 0.2578 1.204331835 0.710231969 1.914563804 62.90% 74.15% 0.47
V49A_Q295L 0.2342 0.2992 0.957090315 0.82428784 1.781378154 53.73% 69.00% 0.37
V49A_S214R 0.2154 0.2196 0.880261545 0.604992011 1.485253555 59.27% 57.53% 0.34
S177W_S214R 0.1776 0.2114 0.725786678 0.582401234 1.308187912 55.48% 50.67% 0.28
Example 7: Generation of ORF2 Variants with Improved Enzymatic Function Using O as Substrate and GPP as Donor Six triple mutants (E09 carrying A53T_M106E_Q161S mutations; D12 carrying A53T_E112D_G205M; C06 carrying Q161A_M162F_Q295A; C11 carrying E112D_L219F_V294F; G12 carrying A17T_Q161W_A232S; and A09 carrying V49A_Q161S_V294A) had improved activity vs. the WT ORF2, based on FIG. 55. Out of these, A09 and E09 were targeted for “breakdown” analysis. For each parental clone targeted six unique mutants are generated (3 doubles and 3 singles). The % enzymatic activity when using O as substrate and GPP as donor was calculated for each of the single and double mutants derived from the breakdown analysis of E09 and A09. FIGS. 78 and 79 depict the % activity for the mutants derived from the breakdown of E09 (FIG. 78); and A09 (FIG. 79) triple mutant clones.
In a similar fashion, breakdown of the other triple mutants—D12, C06, C11 and G12—will be performed; and the % activity for the mutants derived from the breakdown of D12, C06, C11 and G12 will analyzed as described above.
From the analysis of the breakdown mutants of E09 and A09, the following sites were identified as target residues for site saturation: V294, Q161, and A53. Site-saturated mutants of Q161, and A53 described in Examples 1 and 2 were analyzed to determine their % enzymatic activity as compared to WT ORF2 using O as substrate and GPP as donor. These results are depicted in FIG. 80 (Q161) and FIG. 81 (A53). From the results described above, multiple mutations of A53 and Q161 that have significantly higher % activity for producing CBG, as compared to WT ORF2, were identified. Thus, the ORF2 mutants disclosed herein have unexpectedly superior enzymatic functions, in a reaction using O as a substrate and GPP as donor, as compared to WT ORF2.
Site-saturated mutagenesis will be done in the same manner for V294; and % activity will be measured for each of these site saturated mutants.
Example 8: Analysis of ORF2 Mutant Function at pH 5.3 The library of tripleton mutants was screened for the ability to produce CBGA and 5-GOA using OA as substrate and Geranyl pyrophosphate (GPP) as donor at pH 5.3; and this enzymatic activity was compared to that of the WT ORF2. Table 30 provides the summary of the analysis performed on the enzymatic activity of the ORF2 triple mutants to produce CBG and 5-GO using O as substrate and Geranyl pyrophosphate (GPP) as donor.
TABLE 30
5-
GOA
CBGA Pro-
CLONE nMol nMol 5- Total % % Produc- % 5- duc-
ID Mutations CBGA 5-GOA CBGA GOA Product CBGA Activity tion GOA tion
F08 A53T_N173D_ 1.857 0.0154 0.489070319 0.010166359 0.499236677 98.0% 70.98% 0.70 2.04% 0.01
S214R
H02 A53Q_S177W_ 0.7295 0.1064 0.192125362 0.070240296 0.262365658 73.2% 37.30% 0.27 26.77% 0.10
L219F
E09 A53T_M106E_ 0.7007 0.8385 0.184540427 0.553538421 0.738078848 25.0% 104.94% 0.26 75.00% 0.79
Q161S
D12 A53T_E112D_ 0.6102 3.8502 0.16070582 2.541721679 2.7024275 5.9% 384.22% 0.23 94.05% 3.61
G205M
G12 A17T_Q161W_ 0.2702 0.0566 0.071161443 0.037364669 0.108526112 65.6% 15.43% 0.10 34.43% 0.05
A232S
C11 E112D_L219F_ 0.2299 1.1166 0.060547801 0.737127013 0.797674814 7.6% 113.41% 0.09 92.41% 1.05
V294F
D06 A53E_Q161A_ 0.1929 2.8479 0.050803266 1.880050172 1.930853437 2.6% 274.52% 0.07 97.37% 2.67
V294N
H10 M162A_N173D_ 0.1834 2.5826 0.04830129 1.704911539 1.75321283 2.8% 249.26% 0.07 97.24% 2.42
S214F
A09 V49A_Q161S_ 0.1672 0.1311 0.044034764 0.086546079 0.130580843 33.7% 18.57% 0.06 66.28% 0.12
V294A
D04 A53T_D166E_ 0.1279 0.1365 0.033684488 0.090110906 0.123795393 27.2% 17.60% 0.05 72.79% 0.13
Q295W
C06 Q161A_M162F_ 0.0891 0.0334 0.023465894 0.022049115 0.04551501 51.6% 6.47% 0.03 48.44% 0.03
Q295A
H11 A108G_Q161S_ 0.0876 0.12 0.023070845 0.079218379 0.102289224 22.6% 14.54% 0.03 77.45% 0.11
G205M
F09 Q38G_D166E_ 0.0852 0.0626 0.022438767 0.041325588 0.063764355 35.2% 9.07% 0.03 64.81% 0.06
Q295A
C12 N173D_F213M_ 0.0617 10.6898 0.016249671 7.056905202 7.073154873 0.2% 1005.62% 0.02 99.77% 10.03
V294F
A02 Q38G_E112D_ 0.0427 0.4286 0.01124572 0.282941642 0.294187363 3.8% 41.83% 0.02 96.18% 0.40
F123H
D11 F123H_L174V_ 0.0427 0.0282 0.01124572 0.018616319 0.029862039 37.7% 4.25% 0.02 62.34% 0.03
S177E
H05 S177E_S214R_ 0.0367 0.0116 0.009665525 0.007657777 0.017323302 55.8% 2.46% 0.01 44.21% 0.01
R228E
A10 V49S_K118Q_ 0.0302 0.0096 0.007953648 0.00633747 0.014291118 55.7% 2.03% 0.01 44.35% 0.01
S177E
H09 E112G_G205M_ 0.0255 0.22 0.006715828 0.145233694 0.151949523 4.4% 21.60% 0.01 95.58% 0.21
L298W
H03 A17T_F213M_ 0.0215 0.0107 0.005662365 0.007063639 0.012726004 44.5% 1.81% 0.01 55.51% 0.01
S214R
C03 V49L_S214R_ 0.0127 0.009 0.003344746 0.005941378 0.009286124 36.0% 1.32% 0.00 63.98% 0.01
V271E
G04 D227E_R228E_ 0.011 0.0485 0.002897024 0.032017428 0.034914452 8.3% 4.96% 0.00 91.70% 0.05
L298Q
H06 V49L_E112D_ 0.0109 0.0105 0.002870687 0.006931608 0.009802296 29.3% 1.39% 0.00 70.71% 0.01
G286E
H08 M106E_M162A_ 0.0094 0.0425 0.002475639 0.028056509 0.030532148 8.1% 4.34% 0.00 91.89% 0.04
Y216A
G02 A53E_F213M_ 0.0092 0.2181 0.002422965 0.143979403 0.146402369 1.7% 20.81% 0.00 98.34% 0.20
R228Q
B09 C25V_F213M_ 0.0087 0.0575 0.002291283 0.037958806 0.040250089 5.7% 5.72% 0.00 94.31% 0.05
Y216A
G05 A53T_K118N_ 0.0078 0.0297 0.002054253 0.019606549 0.021660802 9.5% 3.08% 0.00 90.52% 0.03
S214F
B08 K118Q_L174V_ 0.0067 0.0208 0.001764551 0.013731186 0.015495737 11.4% 2.20% 0.00 88.61% 0.02
R228Q
E10 E112D_K119A_ 0.0061 0.0176 0.001606531 0.011618696 0.013225227 12.1% 1.88% 0.00 87.85% 0.02
N173D
D08 E112D_K119A_ 0.0056 0.1394 0.001474849 0.09202535 0.093500198 1.6% 13.29% 0.00 98.42% 0.13
N173D
A08 C25V_A232S_ 0.0054 0.0077 0.001422175 0.005083179 0.006505355 21.9% 0.92% 0.00 78.14% 0.01
V271E
G03 L219F_Y283L_ 0.0053 0.0402 0.001395839 0.026538157 0.027933996 5.0% 3.97% 0.00 95.00% 0.04
L298W
G06 K118Q_F123A_ 0.0047 0.0306 0.001237819 0.020200687 0.021438506 5.8% 3.05% 0.00 94.23% 0.03
R228E
H01 K119A_Q161A_ 0.0038 0.0277 0.00100079 0.018286242 0.019287033 5.2% 2.74% 0.00 94.81% 0.03
R228Q
G08 F123W_M162F_ 0.0033 0.0004 0.000869107 0.000264061 0.001133168 76.7% 0.16% 0.00 23.30% 0.00
C209G
B04 A53E_A108G_ 0.0029 0.0075 0.000763761 0.004951149 0.00571491 13.4% 0.81% 0.00 86.64% 0.01
K118N
B10 M106E_Y121W_ 0.0028 0.0102 0.000737424 0.006733562 0.007470986 9.9% 1.06% 0.00 90.13% 0.01
D166E
B06 D166E_S177Y_ 0.0021 0.0146 0.000553068 0.009638236 0.010191304 5.4% 1.45% 0.00 94.57% 0.01
S214F
C02 K118N_K119A_ 0.0021 0.0038 0.000553068 0.002508582 0.00306165 18.1% 0.44% 0.00 81.94% 0.00
V271E
G07 V49S_Y216A_ 0.0021 0.0031 0.000553068 0.002046475 0.002599543 21.3% 0.37% 0.00 78.72% 0.00
V294N
B05 A53Q_Y121W_ 0.0019 0.0081 0.000500395 0.005347241 0.005847636 8.6% 0.83% 0.00 91.44% 0.01
A232S
G10 V49A_Y121W_ 0.0017 0.004 0.000447722 0.002640613 0.003088335 14.5% 0.44% 0.00 85.50% 0.00
C230S
D09 K118N_C209G_ 0.0012 0.0002 0.000316039 0.000132031 0.00044807 70.5% 0.06% 0.00 29.47% 0.00
R228Q
B02 V49A_S177Y_ 0.0009 0.0102 0.000237029 0.006733562 0.006970591 3.4% 0.99% 0.00 96.60% 0.01
C209G
C09 A108G_K119D_ 0.0009 0.0002 0.000237029 0.000132031 0.00036906 64.2% 0.05% 0.00 35.77% 0.00
L298A
B12 A17T_F123W_ 0.0009 0 0.000237029 0 0.000237029 100.0% 0.03% 0.00 0.00% 0.00
L298A
D07 K119A_S214G_ 0.0008 0.4619 0.000210693 0.304924743 0.305135435 0.1% 43.38% 0.00 99.93% 0.43
L298A
G09 M106E_G205L_ 0.0006 0.0001 0.000158019 6.60153E−05 0.000224035 70.5% 0.03% 0.00 29.47% 0.00
C209G
D01 K118Q_Q161W_ 0.0006 0.1332 0.000158019 0.0879324 0.08809042 0.2% 12.52% 0.00 99.82% 0.13
S214F
E08 F123H_L274V_ 0.0003 0.0002 7.90097E−05 0.000132031 0.00021104 37.4% 0.03% 0.00 62.56% 0.00
L298A
The amount of CBGA and 5-GOA produced by each of the ORF2 triple mutants at pH 5.3 was measured using HPLC. FIG. 111 shows the total nMols of prenylated products generated using OA as substrate and GPP as donor by each of the ORF2 mutants, and the proportion of CBGA and 5-GOA within the total prenylated products. An exemplary wild type ORF2 replicate is included in the graph for comparison purposes. The results that show that in comparison with WT ORF2, which has low activity at this pH, some of the triple mutants have enhanced activity at this pH.
FIG. 112 shows the % CBGA and %5-GOA within the total prenylated products produced at pH 5.3 by each of the ORF2 triple mutant clones using OA as substrate and GPP as donor. In this graph, the mutant clones are ordered based on decreasing % CBGA (from left to right) they produce, with the % GO depicted in red. The black dashed line on the graph indicates the % CBGA that is produced by the wild type enzyme.
FIG. 113 shows the ORF2 enzymatic activity (using OA as substrate and GPP as donor) of each of the triple mutant ORF2 clones relative to the wild type enzyme at pH 5.3. % activity was calculated by dividing the nMols of total prenylated products produced by a mutant by the nMols of total prenylated products produced by the wild type control, and expressed as a percentage. The black threshold line is the wild type ORF2% activity. The results show that at least 4 mutants have higher % activity at pH 5.3 compared to WT ORF2.
FIG. 114 shows the CBGA production potential of each of the ORF2 triple mutant clones when using OA as substrate and GPP as donor at pH 5.3. CBGA production potential (interchangeably referred to herein as CBGA production quotient) represents the improvement in CBG production vs. the wild type enzyme. CBGA production potential was calculated by multiplying the % CBGA of the Total Products by the % Activity of each mutant.
In the cluster analysis shown in FIG. 115, high 5-GOA producing mutants at pH 5.3 cluster together towards the right of the graph and high CBGA producing mutants at pH 5.3 cluster towards the left of the graph.
Based on the analysis performed in FIG. 115, the mutants which cluster to the left of the graph were selected for “breakdown” analysis based on the % CBGA produced (Table 31) and % activity (Table 32).
TABLE 31
CBGA
Production Targeted
Rank Clone for
(pH 5.3) ID Mutations Breakdown
1 F08 VA53T_N173D_S214R YES
2 H02 A53Q_S177W_L219F YES
3 E09 A53T_M106E_Q161S YES
5 G12 A17T_Q161W_A232S YES
9 A09 V49A_Q1615_V294A YES
10 D04 A53T_D166E_Q295W YES
11 C06 Q161A_M162F_Q295A YES
12 H11 A108G_Q1615_G205M YES
13 F09 Q38G_D166E_Q295A YES
16 Dll F123H_L174V_S177E YES
17 H05 S177E_S214R_R228E YES
18 A10 V495_K118Q_S177E YES
20 H03 A17T_F213M_S214R YES
TABLE 32
%
Activity Targeted
Rank Clone for
(pH 5.3) ID Mutations Breakdown
1 C12 N173D_F213M_V294F YES
2 D12 A53T_E112D_G205M YES
3 D06 A53E_Q161A_V294N YES
4 H10 M162A_N173D_S214F YES
5 C11 E112D_L219F_V294F YES
Each of these triple mutants listed in Tables 31 and 32 will be broken down to their respective single mutants and double mutants; and all the ORF2 mutants thus generated will be analyzed for their activity at pH 5.3.
Finally, ORF2 mutants carrying point substitutions—Q161H, S214H, and Q295H mutations—were analyzed for their % activity and CBGA production potential at pH 5.3. The results surprisingly show that although WT ORF2 has little to no activity and little to no CBGA production potential at pH 5.3, Q161H and S214H have unexpectedly enhanced activity at this acidic pH. See FIG. 161 and Table 33.
TABLE 33
nMol 5- Total % % CBGA % 5- 5-GOA
Mutations CBGA 5-GOA nMol CBGA GOA Product CBGA Activity Production GOA Production
WT 0.3478 0.9267 0.09159863 0.611763929 0.70336256 13.0% 100.00% 0.13 86.98% 0.87
Q161H 26.7263 0.171 7.038793785 0.11288619 7.151679974 98.4% 1016.78% 10.01 1.58% 0.16
S214H 12.8071 0.0115 3.372952331 0.007591761 3.380544092 99.8% 480.63% 4.80 0.22% 0.01
Q295H 2.0712 2.266 0.545483276 1.49590705 2.041390327 26.7% 290.23% 0.78 73.28% 2.13
Example 9: Amino Acid and Nucleic Acid Sequences Table 34 lists all the amino acid sequences disclosed herein. Sequence information indicates the amino acid substitution of the sequence in comparison to the WT ORF2 (consensus ORF2).
TABLE 34
Amino acid sequences and their SEQ ID NO
SEQ
ID
SEQ info NO Amino Acid Sequence
consensus 1 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
Orf2 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
(NPhB) VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
A108G_Q 2 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
295N_L2 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFGIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
98Q VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVNRGQLKAFDSLED**
K119A_Q 3 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
161S_S17 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKATYAFFPTDNMPGVAELSAIPSMPPA
7E VAENAELFARYGLDKVSMTSMDYKKRQVNLYFEELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
V49S_M1 4 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVSFSMASGRHSTELDFSISVPTSHG
62A_Y28 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
3L VAENAELFARYGLDKVQATSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYLKLGAY
YHITDVQRGLLKAFDSLED**
K118Q_Q 5 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
161W_S2 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFQKTYAFFPTDNMPGVAELSAIPSMPPA
14F VAENAELFARYGLDKVWMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFFV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
Q161A_S 6 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
214G_V2 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
94F VAENAELFARYGLDKVAMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFGV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDFQRGLLKAFDSLED**
F123W_S 7 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
177W_G2 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAWFPTDNMPGVAELSAIPSMPP
86E AVAENAELFARYGLDKVQMTSMDYKKRQVNLYFWELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSF
SVYPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLE
AYYHITDVQRGLLKAFDSLED**
K118N_N 8 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
173D_Q2 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFNKTYAFFPTDNMPGVAELSAIPSMPPA
95N VAENAELFARYGLDKVQMTSMDYKKRQVDLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVNRGLLKAFDSLED**
K119A_Q 9 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
161A_R2 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKATYAFFPTDNMPGVAELSAIPSMPPA
28Q VAENAELFARYGLDKVAMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDQLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
Q38G_E1 10 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFGDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
12D_F12 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGDVTGGFKKTYAHFPTDNMPGVAELSAIPSMPP
3H AVAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFS
VYPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLG
AYYHITDVQRGLLKAFDSLED**
V49A_S1 11 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVAFSMASGRHSTELDFSISVPTSHG
77Y_C20 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
9G VAENAELFARYGLDKVQMTSMDYKKRQVNLYFYELSAQTLEAESVLALVRELGLHVPNELGLKFGKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
K118N_K 12 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
119A_V2 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFNATYAFFPTDNMPGVAELSAIPSMPPA
71E VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLEYGLTLSPKEEYYKLGAY
YHITDVQRGLLKAFDSLED**
L174V_C 13 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
230N_L2 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
98Q VAENAELFARYGLDKVQMTSMDYKKRQVNVYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLNFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGQLKAFDSLED**
K119D_S 14 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
177E_L21 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKDTYAFFPTDNMPGVAELSAIPSMPPA
9F VAENAELFARYGLDKVQMTSMDYKKRQVNLYFEELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTFNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
A53E_E1 15 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMESGRHSTELDFSISVPTSHG
12G_R22 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGGVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
8E VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDELCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
A53E_F2 16 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMESGRHSTELDFSISVPTSHG
13M_R22 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
8Q VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSMS
VYPTLNWETGKIDQLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLG
AYYHITDVQRGLLKAFDSLED**
A53Q_S1 17 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMQSGRHSTELDFSISVPTSHG
77W_L21 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
9F VAENAELFARYGLDKVQMTSMDYKKRQVNLYFWELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFS
VYPTFNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLG
AYYHITDVQRGLLKAFDSLED**
F123A_Y 18 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVLFSMASGRHSTELDFSISVPTSHG
283L DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAAFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYLKLGAY
YHITDVQRGLLKAFDSLED**
C25V_Y1 19 MSEAADVERVYAAMEEAAGLLGVAVARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
21W_M1 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTWAFFPTDNMPGVAELSAIPSMPP
62F AVAENAELFARYGLDKVQFTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFS
VYPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLG
AYYHITDVQRGLLKAFDSLED**
V49L_S2 20 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVLFSMASGRHSTELDFSISVPTSHG
14R_V27 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
1E VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFRV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLEYGLTLSPKEEYYKLGAY
YHITDVQRGLLKAFDSLED**
D227E_C 21 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
230N_Q2 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
95W VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIERLNFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVWRGLLKAFDSLED**
G205L_Y 22 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
216A_C2 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
30S VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELLLKFCKRSFSV
APTLNWETGKIDRLSFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGAY
YHITDVQRGLLKAFDSLED**
Q161S_V 23 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
294F_L29 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
8W VAENAELFARYGLDKVSMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDFQRGWLKAFDSLED**
L219F_Y 24 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
283L_L29 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
8W VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTFNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYLKLGAY
YHITDVQRGWLKAFDSLED**
A17T_F2 25 MSEAADVERVYAAMEETAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
13M_S21 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
4R VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSMR
VYPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLG
AYYHITDVQRGLLKAFDSLED**
L219F_V 26 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
294N_Q2 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
95A VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTFNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDNARGLLKAFDSLED**
A53E_A1 27 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMESGRHSTELDFSISVPTSHG
08G_K11 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFGIDGEVTGGFNKTYAFFPTDNMPGVAELSAIPSMPPA
8N VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
Q38G_E1 28 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFGDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
12G_M16 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGGVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
2F VAENAELFARYGLDKVQFTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
A53T_D1 29 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMTSGRHSTELDFSISVPTSHG
66E_Q29 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
5W VAENAELFARYGLDKVQMTSMEYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVWRGLLKAFDSLED**
L174V_C 30 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
230S_V2 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
94A VAENAELFARYGLDKVQMTSMDYKKRQVNVYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLSFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGAY
YHITDAQRGLLKAFDSLED**
M106E_G 31 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
286E_L29 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSEFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
8W VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLEAY
YHITDVQRGWLKAFDSLED**
D227E_R 32 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
228E_L29 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
8Q VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIEELCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGAY
YHITDVQRGQLKAFDSLED**
M162A_ 33 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
C209G_Y DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
288H VAENAELFARYGLDKVQATSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFGKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
HYHITDVQRGLLKAFDSLED**
A17T_C2 34 MSEAADVERVYAAMEETAGLLGVAVARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
5V_E112 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGGVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
G VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
A53Q_Y1 35 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMQSGRHSTELDFSISVPTSHG
21W_A23 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTWAFFPTDNMPGVAELSAIPSMPP
2S AVAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFS
VYPTLNWETGKIDRLCFSVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
A53Q_S1 36 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMQSGRHSTELDFSISVPTSHG
77Y_Y28 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
8H VAENAELFARYGLDKVQMTSMDYKKRQVNLYFYELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
HYHITDVQRGLLKAFDSLED**
F123H_L 37 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
274V_L2 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAHFPTDNMPGVAELSAIPSMPPA
98A VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGVTLSPKEEYYKLGA
YYHITDVQRGALKAFDSLED**
S177W_S 38 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
214F_Y2 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
83L VAENAELFARYGLDKVQMTSMDYKKRQVNLYFWELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFF
VYPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYLKLGA
YYHITDVQRGLLKAFDSLED**
A53E_G2 39 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMESGRHSTELDFSISVPTSHG
05M_L27 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
4V VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELMLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGVTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
A53T_K1 40 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMTSGRHSTELDFSISVPTSHG
18N_S21 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFNKTYAFFPTDNMPGVAELSAIPSMPPA
4F VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFFV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
5177E_S2 41 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
14R_R22 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
8E VAENAELFARYGLDKVQMTSMDYKKRQVNLYFEELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFRV
YPTLNWETGKIDELCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
K118Q_S 42 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
177Y_D2 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFQKTYAFFPTDNMPGVAELSAIPSMPPA
27E VAENAELFARYGLDKVQMTSMDYKKRQVNLYFYELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIERLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGAY
YHITDVQRGLLKAFDSLED**
D166E_S 43 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
177Y_S2 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
14F VAENAELFARYGLDKVQMTSMEYKKRQVNLYFYELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFFV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
Q161A_ 44 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
M162F_Q DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
295A VAENAELFARYGLDKVAFTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVARGLLKAFDSLED**
A53E_Q1 45 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMESGRHSTELDFSISVPTSHG
61A_V29 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
4N VAENAELFARYGLDKVAMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDNQRGLLKAFDSLED**
A108G_F 46 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
123W_V2 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFGIDGEVTGGFKKTYAWFPTDNMPGVAELSAIPSMPP
94A AVAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFS
VYPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLG
AYYHITDAQRGLLKAFDSLED**
A53Q_G2 47 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMQSGRHSTELDFSISVPTSHG
05L_Q29 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
5N VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELLLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVNRGLLKAFDSLED**
K118Q_F 48 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
123A_R2 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFQKTYAAFPTDNMPGVAELSAIPSMPPA
28E VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDELCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
V49LE_1 49 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVLFSMASGRHSTELDFSISVPTSHG
12D_G28 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGDVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
6E VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLEAY
YHITDVQRGLLKAFDSLED**
G205L_R 50 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
228E_C2 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
30N VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELLLKFCKRSFSV
YPTLNWETGKIDELNFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
M162A_ 51 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
D227E_C DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
230S VAENAELFARYGLDKVQATSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIERLSFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGAY
YHITDVQRGLLKAFDSLED**
V49L_K1 52 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVLFSMASGRHSTELDFSISVPTSHG
19D_G20 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKDTYAFFPTDNMPGVAELSAIPSMPPA
5M VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELMLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
K119A_S 53 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
214G_L2 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKATYAFFPTDNMPGVAELSAIPSMPPA
98A VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFGV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGALKAFDSLED**
Q38G_Y2 54 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFGDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
16A_A23 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
2S VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
APTLNWETGKIDRLCFSVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGAY
YHITDVQRGLLKAFDSLED**
Q161A_V 55 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
271E_Q2 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
95W VAENAELFARYGLDKVAMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLEYGLTLSPKEEYYKLGAY
YHITDVWRGLLKAFDSLED**
V49S_Y2 56 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVSFSMASGRHSTELDFSISVPTSHG
16A_V29 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
4N VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
APTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDNQRGLLKAFDSLED**
F123A_M 57 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
162F_S21 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAAFPTDNMPGVAELSAIPSMPPA
4G VAENAELFARYGLDKVQFTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFGV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
C25V_A2 58 MSEAADVERVYAAMEEAAGLLGVAVARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
32S_V27 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
1E VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFSVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLEYGLTLSPKEEYYKLGAY
YHITDVQRGLLKAFDSLED**
K118Q_L 59 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
174V_R2 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFQKTYAFFPTDNMPGVAELSAIPSMPPA
28Q VAENAELFARYGLDKVQMTSMDYKKRQVNVYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDQLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
V49S_S2 60 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVSFSMASGRHSTELDFSISVPTSHG
14G_V29 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
4A VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFGV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDAQRGLLKAFDSLED**
E1l2D_K 61 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
119A_N1 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGDVTGGFKATYAFFPTDNMPGVAELSAIPSMPPA
73D VAENAELFARYGLDKVQMTSMDYKKRQVDLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
F123W_Q 62 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
161W_D2 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAWFPTDNMPGVAELSAIPSMPP
27E AVAENAELFARYGLDKVWMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFS
VYPTLNWETGKIERLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
A53T_N1 63 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMTSGRHSTELDFSISVPTSHG
73D_S21 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
4R VAENAELFARYGLDKVQMTSMDYKKRQVDLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFRV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
F123W_ 64 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
M162F_C DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAWFPTDNMPGVAELSAIPSMPP
209G AVAENAELFARYGLDKVQFTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFGKRSFS
VYPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLG
AYYHITDVQRGLLKAFDSLED**
M106E_ 65 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
M162A_ DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSEFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
Y216A VAENAELFARYGLDKVQATSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
APTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
V49A_Q1 66 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVAFSMASGRHSTELDFSISVPTSHG
61S_V29 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
4A VAENAELFARYGLDKVSMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDAQRGLLKAFDSLED**
C25V_F2 67 MSEAADVERVYAAMEEAAGLLGVAVARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
13M_Y21 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
6A VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSMS
VAPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLG
AYYHITDVQRGLLKAFDSLED**
A108G_K 68 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
119D_L2 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFGIDGEVTGGFKDTYAFFPTDNMPGVAELSAIPSMPPA
98A VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGALKAFDSLED**
K118N_C 69 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
209G_R2 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFNKTYAFFPTDNMPGVAELSAIPSMPPA
28Q VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFGKRSFSV
YPTLNWETGKIDQLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
A53T_M1 70 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMTSGRHSTELDFSISVPTSHG
06E_Q16 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSEFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
1S VAENAELFARYGLDKVSMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
Q38G_D1 71 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFGDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
66E_Q29 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
5A VAENAELFARYGLDKVQMTSMEYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVARGLLKAFDSLED**
M106E_G 72 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
205L_C2 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSEFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
09G VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELLLKFGKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
E1l2G_G 73 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
205M_L2 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGGVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
98W VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELMLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGWLKAFDSLED**
V49S_K1 74 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVSFSMASGRHSTELDFSISVPTSHG
18Q_S17 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFQKTYAFFPTDNMPGVAELSAIPSMPPA
7E VAENAELFARYGLDKVQMTSMDYKKRQVNLYFEELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
M106E_Y 75 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
121W_D1 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSEFAIDGEVTGGFKKTWAFFPTDNMPGVAELSAIPSMPPA
66E VAENAELFARYGLDKVQMTSMEYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
A53Q_L2 76 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMQSGRHSTELDFSISVPTSHG
74V_Q29 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
5A VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGVTLSPKEEYYKLGA
YYHITDVARGLLKAFDSLED**
V49A_F1 77 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVAFSMASGRHSTELDFSISVPTSHG
23A_Y28 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAAFPTDNMPGVAELSAIPSMPPA
8H VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
HYHITDVQRGLLKAFDSLED**
C25V_F1 78 MSEAADVERVYAAMEEAAGLLGVAVARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
23H_Q29 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAHFPTDNMPGVAELSAIPSMPPA
5N VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVNRGLLKAFDSLED**
K119D_Q 79 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
161W_L2 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKDTYAFFPTDNMPGVAELSAIPSMPPA
98Q VAENAELFARYGLDKVWMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGQLKAFDSLED**
V49A_Y1 80 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVAFSMASGRHSTELDFSISVPTSHG
21W_C23 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTWAFFPTDNMPGVAELSAIPSMPP
0S AVAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFS
VYPTLNWETGKIDRLSFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
M162A_ 81 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
N173D_S DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
214F VAENAELFARYGLDKVQATSMDYKKRQVDLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFFV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
V49L_D1 82 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVLFSMASGRHSTELDFSISVPTSHG
66E_L274 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
V VAENAELFARYGLDKVQMTSMEYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGVTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
V49S_K1 83 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVSFSMASGRHSTELDFSISVPTSHG
19D_F21 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKDTYAFFPTDNMPGVAELSAIPSMPPA
3M VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSMS
VYPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLG
AYYHITDVQRGLLKAFDSLED**
E112D_L 84 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
219F_V2 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGDVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
94F VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTFNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDFQRGLLKAFDSLED**
F123H_L 85 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
174V_S1 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAHFPTDNMPGVAELSAIPSMPPA
77E VAENAELFARYGLDKVQMTSMDYKKRQVNVYFEELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
E1l2G_F 86 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
123H_S2 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGGVTGGFKKTYAHFPTDNMPGVAELSAIPSMPP
14G AVAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFG
VYPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLG
AYYHITDVQRGLLKAFDSLED**
F123A_S 87 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
177W_G2 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAAFPTDNMPGVAELSAIPSMPPA
05L VAENAELFARYGLDKVQMTSMDYKKRQVNLYFWELSAQTLEAESVLALVRELGLHVPNELLLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
S177W_Y 88 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
288H_V2 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
94N VAENAELFARYGLDKVQMTSMDYKKRQVNLYFWELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFS
VYPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLG
AHYHITDNQRGLLKAFDSLED**
A108G_Q 89 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
161S_G2 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFGIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
05M VAENAELFARYGLDKVSMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELMLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
Y121W_S 90 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
177Y_G2 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTWAFFPTDNMPGVAELSAIPSMPP
86E AVAENAELFARYGLDKVQMTSMDYKKRQVNLYFYELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFS
VYPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLEA
YYHITDVQRGLLKAFDSLED**
A17T_F1 91 MSEAADVERVYAAMEETAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
23W_L29 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAWFPTDNMPGVAELSAIPSMPP
8A AVAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFS
VYPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLG
AYYHITDVQRGALKAFDSLED**
N173D_F 92 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
213M_V2 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
94F VAENAELFARYGLDKVQMTSMDYKKRQVDLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSMS
VYPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLG
AYYHITDFQRGLLKAFDSLED**
A53T_E1 93 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMTSGRHSTELDFSISVPTSHG
12D_G20 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGDVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
5M VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELMLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
Q38G_L1 94 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFGDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
74V_S21 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
4R VAENAELFARYGLDKVQMTSMDYKKRQVNVYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFRV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
A17T_V4 95 MSEAADVERVYAAMEETAGLLGVACARDKIYPLLSTFQDTLVEGGSVVAFSMASGRHSTELDFSISVPTSHG
9A_C230 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
N VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLNFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
A17T_Q1 96 MSEAADVERVYAAMEETAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
61W_A23 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
2S VAENAELFARYGLDKVWMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFSVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGAY
YHITDVQRGLLKAFDSLED**
Q161W_ 97 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
R228Q_Q DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
295W VAENAELFARYGLDKVWMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDQLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVWRGLLKAFDSLED**
Y288A 98 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
AYHITDVQRGLLKAFDSLED**
Y288C 99 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
CYHITDVQRGLLKAFDSLED**
Y288D 100 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
DYHITDVQRGLLKAFDSLED**
Y288E 101 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGAE
YHITDVQRGLLKAFDSLED**
Y288F 102 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGAF
YHITDVQRGLLKAFDSLED**
Y288G 103 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
GYHITDVQRGLLKAFDSLED**
Y288H 104 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
HYHITDVQRGLLKAFDSLED**
Y288I 105 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGAI
YHITDVQRGLLKAFDSLED**
Y288K 106 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
KYHITDVQRGLLKAFDSLED**
Y288L 107 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGAL
YHITDVQRGLLKAFDSLED**
Y288M 108 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
MYHITDVQRGLLKAFDSLED**
Y288N 109 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
NYHITDVQRGLLKAFDSLED**
Y288P 110 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGAP
YHITDVQRGLLKAFDSLED**
Y288Q 111 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
QYHITDVQRGLLKAFDSLED**
Y288R 112 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
RYHITDVQRGLLKAFDSLED**
Y288S 113 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGAS
YHITDVQRGLLKAFDSLED**
Y288T 114 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGAT
YHITDVQRGLLKAFDSLED**
Y288V 115 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
VYHITDVQRGLLKAFDSLED**
Y288W 116 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
WYHITDVQRGLLKAFDSLED**
Y288Y 117 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
Q161A 118 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVAMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
Q161C 119 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVCMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
Q161D 120 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVDMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
Q161E 121 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVEMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
Q161F 122 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVFMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
Q161G 123 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVGMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
Q161H 124 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVHMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
Q161I 125 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVIMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
Q161K 126 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVKMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
Q161L 127 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVLMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
Q161M 128 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVMMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
Q161N 129 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVNMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
Q161P 130 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVPMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
Q161Q 131 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
Q161R 132 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVRMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
Q161S 133 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVSMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
Q161T 134 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVTMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
Q161V 135 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVVMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
Q161W 136 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVWMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
Q161Y 137 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVYMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
Q295A 138 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVARGLLKAFDSLED**
Q295C 139 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVCRGLLKAFDSLED**
Q295D 140 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVDRGLLKAFDSLED**
Q295E 141 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVERGLLKAFDSLED**
Q295F 142 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVFRGLLKAFDSLED**
Q295G 143 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVGRGLLKAFDSLED**
Q295H 144 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVHRGLLKAFDSLED**
Q295I 145 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVIRGLLKAFDSLED**
Q295K 146 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVKRGLLKAFDSLED**
Q295L 147 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVLRGLLKAFDSLED**
Q295M 148 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVMRGLLKAFDSLED**
Q295N 149 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVNRGLLKAFDSLED**
Q295P 150 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVPRGLLKAFDSLED**
Q295Q 151 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
Q295R 152 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVRRGLLKAFDSLED**
Q295S 153 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVSRGLLKAFDSLED**
Q295T 154 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVTRGLLKAFDSLED**
Q295V 155 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVVRGLLKAFDSLED**
Q295W 156 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVWRGLLKAFDSLED**
Q295Y 157 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVYRGLLKAFDSLED**
S214A 158 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFAV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
S214C 159 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFCV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
S214D 160 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFDV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
S214E 161 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFEV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
S214F 162 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFFV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
S214G 163 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFGV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
S214H 164 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFHV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
S214I 165 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFIV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
S214K 166 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFKV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
S214L 167 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFLV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
S214M 168 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFM
VYPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLG
AYYHITDVQRGLLKAFDSLED**
S214N 169 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFNV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
S214P 170 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFPV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
S214Q 171 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFQV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVORGLLKAFDSLED**
S214R 172 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFRV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
S214S 173 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
S214T 174 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFTV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
S214V 175 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFVV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
S214W 176 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFW
VYPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLG
AYYHITDVQRGLLKAFDSLED**
S214Y 177 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFYV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
A53A 178 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
A53C 179 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMCSGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
A53D 180 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMDSGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
A53E 181 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMESGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
A53F 182 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMFSGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
A53G 183 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMGSGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
A53H 184 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMHSGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
A53I 185 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMISGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
A53K 186 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMKSGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
A53L 187 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMLSGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
A53M 188 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMMSGRHSTELDFSISVPTSH
GDPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPP
AVAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFS
VYPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLG
AYYHITDVQRGLLKAFDSLED**
A53N 189 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMNSGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
A53P 190 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMPSGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
A53Q 191 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMQSGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
A53R 192 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMRSGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
A53S 193 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMSSGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
A53T 194 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMTSGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
A53V 195 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMVSGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
A53W 196 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMWSGRHSTELDFSISVPTSH
GDPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPP
AVAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFS
VYPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLG
AYYHITDVQRGLLKAFDSLED**
A53Y 197 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMYSGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
V294N_ 198 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDNQRGLLKAFDSLED**
L219F_V 199 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
294N DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTFNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDNQRGLLKAFDSLED**
L219F_Q 200 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
295A DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTFNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVARGLLKAFDSLED**
V294N_Q 201 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
295A DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDNARGLLKAFDSLED**
L219F 202 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTFNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
V49A 203 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVAFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
V294A 204 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDAQRGLLKAFDSLED**
V49A_V2 205 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVAFSMASGRHSTELDFSISVPTSHG
94A DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDAQRGLLKAFDSLED**
Q161S_V 206 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
294A DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVSMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDAQRGLLKAFDSLED**
V49A_Q1 207 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVAFSMASGRHSTELDFSISVPTSHG
61S DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVSMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
S177Y 208 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFYELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
Y288H 209 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
HYHITDVQRGLLKAFDSLED**
A53Q_Y2 210 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMQSGRHSTELDFSISVPTSHG
88H DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
HYHITDVQRGLLKAFDSLED**
S177Y_Y 211 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
288H DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFYELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
HYHITDVQRGLLKAFDSLED**
A53Q_S1 212 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMQSGRHSTELDFSISVPTSHG
77Y DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFYELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
D166E 213 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMEYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
A53T_Q2 214 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMTSGRHSTELDFSISVPTSHG
95W DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVWRGLLKAFDSLED**
D166E_Q 215 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
295W DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMEYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVWRGLLKAFDSLED**
A53T_ 216 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMTSGRHSTELDFSISVPTSHG
D166E DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMEYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
F123H 217 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAHFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
L174V 218 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNVYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
S177E 219 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFEELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
F123H_S 220 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
177E DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAHFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFEELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
L174V_S 221 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
177E DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNVYFEELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
F123H_L 222 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
174V DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAHFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNVYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
M106E 223 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSEFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
A53T_Q1 224 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMTSGRHSTELDFSISVPTSHG
61S DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVSMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
M106E_Q 225 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
161S DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSEFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVSMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
A53T_M1 226 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMTSGRHSTELDFSISVPTSHG
06E DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSEFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
Q38G 227 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFGDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
D166E 228 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMEYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
Q295A 229 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVARGLLKAFDSLED**
Q38G_Q2 230 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFGDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
95A DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVARGLLKAFDSLED**
D166E_Q 231 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
295A DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMEYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVARGLLKAFDSLED**
Q38G_D1 232 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFGDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
66E DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMEYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
K118N 233 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFNKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
S214F 234 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFFV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
A53T_S2 235 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMTSGRHSTELDFSISVPTSHG
14F DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFFV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
K118N_S 236 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
214F DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFNKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFFV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
A53T_K1 237 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMTSGRHSTELDFSISVPTSHG
18N DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFNKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
S177W 238 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFWELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFS
VYPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLG
AYYHITDVQRGLLKAFDSLED**
L219F_ 239 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
S177W_L DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTFNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
219F 240 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFWELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFS
VYPTFNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLG
AYYHITDVQRGLLKAFDSLED**
A53Q_S1 241 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMQSGRHSTELDFSISVPTSHG
77W DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFWELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFS
VYPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLG
AYYHITDVQRGLLKAFDSLED**
A53Q_L2 242 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMQSGRHSTELDFSISVPTSHG
19F DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTFNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
A108G 243 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFGIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
G205M 244 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELMLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
A108G_G 245 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
205M DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFGIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELMLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
Q161S_G 246 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
205M DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVSMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELMLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
A108G_Q 247 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
161S DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFGIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVSMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
K118Q 248 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFQKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
M162A 249 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQATSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
Y283L 250 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYLKLGAY
YHITDVQRGLLKAFDSLED**
V271E 251 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLEYGLTLSPKEEYYKLGAY
YHITDVQRGLLKAFDSLED**
K119D 252 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKDTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
L274V 253 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGVTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
A232S 254 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFSVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGAY
YHITDVQRGLLKAFDSLED**
Q161A 255 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVAMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
G205L 256 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELLLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
S214G 257 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFGV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
Y216A 258 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
APTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
N173D 259 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVDLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
F123W 260 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAWFPTDNMPGVAELSAIPSMPP
AVAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFS
VYPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLG
AYYHITDVQRGLLKAFDSLED**
K118N 261 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFNKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
R228Q 262 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDQLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
Q161W 263 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVWMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
V294F 264 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDFQRGLLKAFDSLED**
S214R 265 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFRV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
G286E 266 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLEAY
YHITDVQRGLLKAFDSLED**
R228E 267 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDELCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
C230S 268 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLSFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGAY
YHITDVQRGLLKAFDSLED**
D227E 269 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIERLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGAY
YHITDVQRGLLKAFDSLED**
M162F 270 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQFTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
F123A 271 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAAFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
A53T_V2 272 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMTSGRHSTELDFSISVPTSHG
94A DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDAQRGLLKAFDSLED**
A53T_Q1 273 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMTSGRHSTELDFSISVPTSHG
61S_V29 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
4A VAENAELFARYGLDKVSMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDAQRGLLKAFDSLED**
A53T_Q1 274 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMTSGRHSTELDFSISVPTSHG
61S_V29 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
4N VAENAELFARYGLDKVSMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDNQRGLLKAFDSLED**
A53T_Q2 275 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMTSGRHSTELDFSISVPTSHG
95A DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVARGLLKAFDSLED**
Q161S_V 276 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
294A_Q2 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
95A VAENAELFARYGLDKVSMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDAARGLLKAFDSLED**
A53T_Q1 277 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMTSGRHSTELDFSISVPTSHG
61S_Q29 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
5A VAENAELFARYGLDKVSMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVARGLLKAFDSLED**
A53T_V2 278 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMTSGRHSTELDFSISVPTSHG
94A_Q29 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
5A VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDAARGLLKAFDSLED**
A53T_Q1 279 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMTSGRHSTELDFSISVPTSHG
61S_V29 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
4A_Q295 VAENAELFARYGLDKVSMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
A YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDAARGLLKAFDSLED**
A53T_Q1 280 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMTSGRHSTELDFSISVPTSHG
61S_V29 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
4N_Q295 VAENAELFARYGLDKVSMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
A YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDNARGLLKAFDSLED**
A53T_Q2 281 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMTSGRHSTELDFSISVPTSHG
95W DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVWRGLLKAFDSLED**
Q161S_V 282 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
294A_Q2 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
95W VAENAELFARYGLDKVSMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDAWRGLLKAFDSLED**
A53T_Q1 283 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMTSGRHSTELDFSISVPTSHG
61S_Q29 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
5W VAENAELFARYGLDKVSMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVWRGLLKAFDSLED**
A53T_V2 284 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMTSGRHSTELDFSISVPTSHG
94A_Q29 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
5W VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDAWRGLLKAFDSLED**
A53T_Q1 285 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMTSGRHSTELDFSISVPTSHG
61S_V29 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
4A_Q295 VAENAELFARYGLDKVSMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
W YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDAWRGLLKAFDSLED**
A53T_Q1 286 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMTSGRHSTELDFSISVPTSHG
61S_V29 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
4N_Q295 VAENAELFARYGLDKVSMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
W YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDNWRGLLKAFDSLED**
S177W_Q 287 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
295A DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFWELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFS
VYPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLG
AYYHITDVARGLLKAFDSLED**
S177W_S 288 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
214R DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFWELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFR
VYPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLG
AYYHITDVQRGLLKAFDSLED**
Q161S_S 289 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
177W DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVSMTSMDYKKRQVNLYFWELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
A53T_S1 290 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMTSGRHSTELDFSISVPTSHG
77W DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFWELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFS
VYPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLG
AYYHITDVQRGLLKAFDSLED**
V49A_Q2 291 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVAFSMASGRHSTELDFSISVPTSHG
95L DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVLRGLLKAFDSLED**
V49A_S2 292 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVAFSMASGRHSTELDFSISVPTSHG
14R DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFRV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
A53T_Q2 293 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMTSGRHSTELDFSISVPTSHG
95F DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVFRGLLKAFDSLED**
A53T_S2 294 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMTSGRHSTELDFSISVPTSHG
14R DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFRV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
A53T_Q1 295 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMTSGRHSTELDFSISVPTSHG
61S DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVSMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
Q161S_Q 296 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
295F DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVSMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVFRGLLKAFDSLED**
Q161S_Q 297 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
295L DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVSMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVLRGLLKAFDSLED**
Q161S_S 298 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
214R DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVSMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFRV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
S214R_Q 299 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVVFSMASGRHSTELDFSISVPTSHG
295F DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVQMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFRV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVFRGLLKAFDSLED**
V49A_Q1 300 MSEAADVERVYAAMEEAAGLLGVACARDKIYPLLSTFQDTLVEGGSVVAFSMASGRHSTELDFSISVPTSHG
615 DPYATVVEKGLFPATGHPVDDLLADTQKHLPVSMFAIDGEVTGGFKKTYAFFPTDNMPGVAELSAIPSMPPA
VAENAELFARYGLDKVSMTSMDYKKRQVNLYFSELSAQTLEAESVLALVRELGLHVPNELGLKFCKRSFSV
YPTLNWETGKIDRLCFAVISNDPTLVPSSDEGDIEKFHNYATKAPYAYVGEKRTLVYGLTLSPKEEYYKLGA
YYHITDVQRGLLKAFDSLED**
Table 35 lists all the nucleic acid sequences disclosed herein. The protein sequence (WT or mutant ORF2) encoded by the respective nucleic acid sequence is provided in Column 1 of Table 35.
TABLE 35
Nucleic acid sequences and their SEQ ID NO
SEQ
SEQ ID
info NO DNA Sequence
Consensus 301 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
Orf2 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
(NphB) TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
A108G_ 302 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
Q295 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
N_L29 TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
8Q TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGGTATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTAACCGTGGCCAGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
K119A_ 303 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
Q161 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
S_S177 TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
E TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAGCGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGAGCATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCGAGGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
V49S_ 304 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
M162A_ TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCA
Y283 GCTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
L TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGGCGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACCTGAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
K118Q_ 305 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
Q161 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
W_S21 TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
4F TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCCAGAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGTGGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTTTCGTTTACCCGACCC
TGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Q161A_ 306 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
S214 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
G_V29 TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
4F TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGGCGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTGGTGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACTTCCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
F123W_ 307 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
S177 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
W_G2 TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
86E TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTGGTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCTGGGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGAGGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
K118N_ 308 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
N173 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
D_Q29 TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
5N TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAACAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAGACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTAACCGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
K119A_ 309 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
Q161 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
A_R22 TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
8Q TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAGCGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGGCGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCAGCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCG
AGCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAG
AAGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCAC
ATTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Q38G_ 310 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
E112D_ TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTGGTGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
F123 TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
H TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGACGTCACGGGCGGTTTCAAAAAGACCTAT
GCTCACTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
V49A_ 311 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
S177Y_ TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
C209 CGTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
G TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCTACGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCGGTAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
K118N_ 312 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
K119 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
A_V27 TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
1E TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAACGCGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGAGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
L174V_ 313 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
C230 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
N_L29 TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
8Q TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACGTTTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGAACTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCG
AGCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAG
AAGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCAC
ATTACCGACGTTCAACGTGGCCAGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
K119D_ 314 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
S177 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
E_L21 TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
9F TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAGACACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCGAGGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
TTCAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
A53E_ tag ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
E112G_ TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
R228 TTTTTAGCATGGAGAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
E TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGGTGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACGAGCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCG
AGCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAG
AAGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCAC
ATTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
A53E_ 316 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
F213M_ TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
R228 TTTTTAGCATGGAGAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
Q TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCATGAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCAGCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCG
AGCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAG
AAGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCAC
ATTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
A53Q_ 317 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
S177W_ TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
L219 TTTTTAGCATGCAGAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
F TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCTGGGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
TTCAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
F123A_ 318 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
Y283 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCC
L TGTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTGCGTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACCTGAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
C25V_ 319 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
Y121 GTTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
W_M1 TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
62F TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTGG
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGTTCACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
V49L_ 320 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
S214R_ TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCC
V271 TGTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
E TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTCGTGTTTACCCGACCC
TGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGAGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
D227E_ 321 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
C230 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
N_Q29 TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
5W TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGAGCGCCTGAACTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCG
AGCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAG
AAGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCAC
ATTACCGACGTTTGGCGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
G205L_ 322 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
Y216 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
A_C23 TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
0S TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGCTGCTGAAATTCTGCAAGCGTAGCTTTAGCGTTGCGCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGAGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCG
AGCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAG
AAGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCAC
ATTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Q161S_ 323 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
V294 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
F_L29 TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
8W TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGAGCATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACTTCCAACGTGGCTGGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
L219F_ 324 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
Y283 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
L_L29 TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
8W TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
TTCAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACCTGAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCTGGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
A17T_ 325 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAACCGCCGGTTTGCTGGGTGTGGCT
F213M_ TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
S214 TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
R TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCATGCGTGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
L219F_ 326 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
V294 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
N_Q29 TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
5A TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
TTCAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACAACGCGCGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
A53E_ 327 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
A108G_ TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
K118 TTTTTAGCATGGAGAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
N TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGGTATTGATGGTGAGGTCACGGGCGGTTTCAACAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Q38G_ 328 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
E112G_ TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTGGTGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
M162 TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
F TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGGTGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGTTCACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
A53T_ 329 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
D166E_ TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
Q295 TTTTTAGCATGACCAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
W TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGAGTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTTGGCGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
L174V_ 330 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
C230 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
S_V29 TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
4A TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACGTTTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGAGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCG
AGCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAG
AAGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCAC
ATTACCGACGCGCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
M106E_ 331 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
G286 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
E_L29 TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
8W TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCGAGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGAGGCGTACTACCACA
TTACCGACGTTCAACGTGGCTGGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
D227E_ 332 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
R228 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
E_L29 TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
8Q TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGAGGAGCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCG
AGCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAG
AAGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCAC
ATTACCGACGTTCAACGTGGCCAGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
M162A_ 333 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
C209 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
G_Y28 TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
8H TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGGCGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCGGTAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGCACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
A17T_ 334 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAACCGCCGGTTTGCTGGGTGTGGCT
C25V_ GTTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
E112G TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGGTGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
A53Q_ 335 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
Y121 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
W_A2 TTTTTAGCATGCAGAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
32S TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTGG
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTAGCGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
A53Q_ 336 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
S177Y_ TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
Y288 TTTTTAGCATGCAGAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
H TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCTACGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGCACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
F123H_ 337 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
L274 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
V_L29 TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
8A TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTCACTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTGTTACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCGCGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
S177W_ 338 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
S214F_ TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
Y283 TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
L TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCTGGGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTTTCGTTTACCCGACCC
TGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACCTGAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
A53E_ 339 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
G205M_ TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
L274 TTTTTAGCATGGAGAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
V TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGATGCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTGTTACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
A53T_ 340 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
K118N_ TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
S214F TTTTTAGCATGACCAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAACAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTTTCGTTTACCCGACCC
TGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
S177E_ 341 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
S214 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
R_R22 TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
8E TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCGAGGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTCGTGTTTACCCGACCC
TGAACTGGGAAACCGGCAAGATTGACGAGCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
K118Q_ 342 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
S177 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
Y_D22 TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
7E TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCCAGAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCTACGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGAGCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCG
AGCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAG
AAGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCAC
ATTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
D166E_ 343 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
S177 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
Y_S21 TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
4F TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGAGTACAAGAAAC
GCCAGGTAAACCTGTATTTCTACGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTTTCGTTTACCCGACCC
TGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Q161A_ 344 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
M162 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
F_Q29 TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
5A TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGGCGTTCACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTGCGCGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
A53E_ 345 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
Q161A_ TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
V294 TTTTTAGCATGGAGAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
N TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGGCGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACAACCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
A108G_ 346 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
F123 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
W_V2 TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
94A TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGGTATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTGGTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGCGCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
A53Q_ 347 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
G205L_ TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
Q295 TTTTTAGCATGCAGAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
N TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGCTGCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTAACCGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
K118Q_ 348 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
F123 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
A_R22 TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
8E TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCCAGAAGACCTAT
GCTGCGTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACGAGCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCG
AGCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAG
AAGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCAC
ATTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
V49L_ 349 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
E112D_ TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCC
G286 TGTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
E TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGACGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGAGGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
G205L_ 350 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
R228 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
E_C23 TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
0N TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGCTGCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACGAGCTGAACTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCG
AGCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAG
AAGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCAC
ATTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
M162A_ 351 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
D227 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
E_C23 TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
0S TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGGCGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGAGCGCCTGAGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCG
AGCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAG
AAGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCAC
ATTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
V49L_ 352 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
K119D_ TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCC
G205 TGTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
M TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAGACACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGATGCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
K119A_ 353 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
S214 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
G_L29 TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
8A TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAGCGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTGGTGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCGCGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Q38G_ 354 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
Y216A_ TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTGGTGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
A232 TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
S TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTGCGCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTAGCGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Q161A_ 355 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
V271 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
E_Q29 TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
5W TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGGCGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGAGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTTGGCGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
V49S_ 356 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
Y216A_ TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCA
V294 GCTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
N TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTGCGCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACAACCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
F123A_ 357 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
M162 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
F_S214 TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
G TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTGCGTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGTTCACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTGGTGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
C25V_ 358 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
A232S_ GTTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
V271 TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
E TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTAGCGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGAGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
K118Q_ 359 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
L174 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
V_R22 TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
8Q TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCCAGAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACGTTTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCAGCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCG
AGCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAG
AAGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCAC
ATTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
V49S_ 360 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
S214G_ TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCA
V294 GCTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
A TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTGGTGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGCGCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
E112D_ 361 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
K119 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
A_N17 TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
3D TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGACGTCACGGGCGGTTTCAAAGCGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAGACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
F123W_ 362 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
Q161 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
W_D2 TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
27E TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTGGTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGTGGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGAGCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCG
AGCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAG
AAGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCAC
ATTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
A53T_ 363 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
N173D_ TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
S214 TTTTTAGCATGACCAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
R TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAGACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTCGTGTTTACCCGACCC
TGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
F123W_ 364 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
M162 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
F_C20 TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
9G TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTGGTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGTTCACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCGGTAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
M106E_ 365 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
M162 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
A_Y21 TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
6A TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCGAGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGGCGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTGCGCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
V49A_ 366 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
Q161S_ TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
V294 CGTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
A TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGAGCATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGCGCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
C25V_ 367 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
F213M_ GTTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
Y216 TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
A TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCATGAGCGTTGCGCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
A108G_ 368 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
K119 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
D_L29 TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
8A TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGGTATTGATGGTGAGGTCACGGGCGGTTTCAAAGACACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCGCGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
K118N_ 369 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
C209 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
G_R22 TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
8Q TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAACAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCGGTAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCAGCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCG
AGCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAG
AAGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCAC
ATTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
A53T_ 370 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
M106E_ TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
Q161 TTTTTAGCATGACCAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
S TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCGAGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGAGCATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Q38G_ 371 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
D166E_ TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTGGTGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
Q295 TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
A TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGAGTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTGCGCGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
M106E_ 372 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
G205 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
L_C20 TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
9G TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCGAGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGCTGCTGAAATTCGGTAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
E112G_ 373 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
G205 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
M_L29 TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
8W TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGGTGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGATGCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCTGGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
V49S_ 374 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
K118Q_ TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCA
S177 GCTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
E TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCCAGAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCGAGGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
M106E_ 375 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
Y121 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
W_D1 TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
66E TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCGAGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTGG
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGAGTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
A53Q_ 376 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
L274V_ TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
Q295 TTTTTAGCATGCAGAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
A TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTGTTACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTGCGCGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
V49A_ 377 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
F123A_ TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
Y288 CGTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
H TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTGCGTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGCACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
C25V_ 378 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
F123H_ GTTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
Q295 TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
N TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTCACTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTAACCGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
K119D_ 379 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
Q161 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
W_L29 TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
8Q TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAGACACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGTGGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCAGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
V49A_ 380 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
Y121 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
W_C23 CGTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
0S TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTGG
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGAGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCG
AGCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAG
AAGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCAC
ATTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
M162A_ 381 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
N173 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
D_S21 TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
4F TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGGCGACCAGCATGGATTACAAGAAAC
GCCAGGTAGACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTTTCGTTTACCCGACCC
TGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
V49L_ 382 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
D166E_ TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCC
L274 TGTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
V TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGAGTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTGTTACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
V49S_ 383 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
K119D_ TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCA
F213 GCTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
M TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAGACACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCATGAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
E112D_ 384 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
L219 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
F_V29 TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
4F TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGACGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
TTCAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACTTCCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
F123H_ 385 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
L174 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
V_S17 TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
7E TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTCACTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACGTTTATTTCGAGGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
E112G_ 386 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
F123 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
H_S21 TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
4G TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGGTGTCACGGGCGGTTTCAAAAAGACCTAT
GCTCACTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTGGTGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
F123A_ 387 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
S177 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
W_G2 TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
05L TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTGCGTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCTGGGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGCTGCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
S177W_ 388 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
Y288 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
H_V29 TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
4N TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCTGGGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGCACTACCACA
TTACCGACAACCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
A108G_ 389 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
Q161 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
S_G20 TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
5M TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGGTATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGAGCATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGATGCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Y121 390 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
W_S17 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
7YG2 TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
86E TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTGG
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCTACGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGAGGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
A17T_ 391 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAACCGCCGGTTTGCTGGGTGTGGCT
F123W_ TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
L298 TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
A TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTGGTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCGCGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
N173D_ 392 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
F213 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
M_V29 TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
4F TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAGACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCATGAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACTTCCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
A53T_ 393 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
E112D_ TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
G205 TTTTTAGCATGACCAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
M TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGACGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGATGCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Q38G_ 394 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
L174V_ TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTGGTGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
S214 TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
R TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACGTTTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTCGTGTTTACCCGACCC
TGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
A17T_ 395 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAACCGCCGGTTTGCTGGGTGTGGCT
V49A_ TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
C230N CGTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGAACTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCG
AGCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAG
AAGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCAC
ATTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
A17T_ 396 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAACCGCCGGTTTGCTGGGTGTGGCT
Q161 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
W_A2 TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
32S TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGTGGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTAGCGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Q161 397 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
W_R22 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
8Q_Q2 TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
95W TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGTGGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCAGCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCG
AGCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAG
AAGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCAC
ATTACCGACGTTTGGCGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Y288A 398 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGgcaTACCACAT
TACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Y288C 399 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGtgtTACCACATT
ACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Y288D 400 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGgatTACCACAT
TACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Y288E 401 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGgaaTACCACAT
TACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Y288F 402 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGtttTACCACATT
ACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Y288G 403 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGggtTACCACAT
TACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Y288H 404 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGcatTACCACATT
ACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Y288I 405 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGattTACCACATT
ACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Y288K 406 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGaaaTACCACAT
TACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Y288L 407 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGctgTACCACAT
TACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Y288M 408 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGatgTACCACAT
TACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Y288N 409 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGaatTACCACATT
ACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Y288P 410 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGccgTACCACAT
TACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Y288Q 411 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGcagTACCACAT
TACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Y288R 412 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGcgtTACCACAT
TACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Y288S 413 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGagcTACCACAT
TACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Y288T 414 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGaccTACCACAT
TACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Y288V 415 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGgttTACCACATT
ACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Y288W 416 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGtggTACCACAT
TACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Y288Y 417 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGtatTACCACATT
ACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Q161A 418 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGgcaATGACCAGCATGGATTACAAGAAACG
CCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCGT
GAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACCC
TGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Q161C 419 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGtgtATGACCAGCATGGATTACAAGAAACGC
CAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCGTG
AGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACCCT
GAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGAG
CAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGAA
GCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACAT
TACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Q161D 420 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGgatATGACCAGCATGGATTACAAGAAACG
CCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCGT
GAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACCC
TGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Q161E 421 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGgaaATGACCAGCATGGATTACAAGAAACG
CCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCGT
GAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACCC
TGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Q161F 422 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGtttATGACCAGCATGGATTACAAGAAACGC
CAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCGTG
AGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACCCT
GAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGAG
CAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGAA
GCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACAT
TACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Q161G 423 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGggtATGACCAGCATGGATTACAAGAAACG
CCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCGT
GAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACCC
TGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Q161H 424 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGcatATGACCAGCATGGATTACAAGAAACGC
CAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCGTG
AGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACCCT
GAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGAG
CAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGAA
GCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACAT
TACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Q161I 425 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGattATGACCAGCATGGATTACAAGAAACGC
CAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCGTG
AGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACCCT
GAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGAG
CAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGAA
GCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACAT
TACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Q161K 426 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGaaaATGACCAGCATGGATTACAAGAAACG
CCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCGT
GAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACCC
TGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Q161L 427 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGctgATGACCAGCATGGATTACAAGAAACG
CCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCGT
GAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACCC
TGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Q161M 428 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGatgATGACCAGCATGGATTACAAGAAACG
CCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCGT
GAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACCC
TGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Q161N 429 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGaatATGACCAGCATGGATTACAAGAAACGC
CAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCGTG
AGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACCCT
GAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGAG
CAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGAA
GCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACAT
TACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Q161P 430 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGccgATGACCAGCATGGATTACAAGAAACG
CCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCGT
GAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACCC
TGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Q161Q 431 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGcagATGACCAGCATGGATTACAAGAAACG
CCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCGT
GAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACCC
TGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Q161R 432 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGcgtATGACCAGCATGGATTACAAGAAACG
CCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCGT
GAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACCC
TGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Q161S 433 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGagcATGACCAGCATGGATTACAAGAAACG
CCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCGT
GAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACCC
TGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Q161T 434 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGaccATGACCAGCATGGATTACAAGAAACG
CCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCGT
GAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACCC
TGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Q161V 435 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGgttATGACCAGCATGGATTACAAGAAACGC
CAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCGTG
AGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACCCT
GAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGAG
CAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGAA
GCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACAT
TACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Q161W 436 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGtggATGACCAGCATGGATTACAAGAAACG
CCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCGT
GAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACCC
TGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Q161Y 437 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGtatATGACCAGCATGGATTACAAGAAACGC
CAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCGTG
AGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACCCT
GAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGAG
CAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGAA
GCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACAT
TACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Q295A 438 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTgcaCGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Q295C 439 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTtgtCGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Q295D 440 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTgatCGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Q295E 441 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTgaaCGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Q295F 442 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTtttCGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Q295G 443 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTggtCGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Q295H 444 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTcatCGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Q295I 445 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTattCGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Q295K 446 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTaaaCGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Q295L 447 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTctgCGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Q295M 448 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTatgCGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Q295N 449 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTaatCGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Q295P 450 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTccgCGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Q295Q 451 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTcagCGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Q295R 452 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTcgtCGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Q295S 453 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTagcCGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Q295T 454 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTaccCGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Q295V 455 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTgttCGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Q295W 456 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTtggCGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Q295Y 457 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTtatCGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
S214A 458 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTgcaGTTTACCCGACCCT
GAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGAG
CAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGAA
GCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACAT
TACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
S214C 459 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTtgtGTTTACCCGACCCT
GAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGAG
CAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGAA
GCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACAT
TACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
S214D 460 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTgatGTTTACCCGACCCT
GAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGAG
CAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGAA
GCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACAT
TACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
S214E 461 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTgaaGTTTACCCGACCCT
GAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGAG
CAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGAA
GCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACAT
TACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
S214F 462 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTtttGTTTACCCGACCCT
GAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGAG
CAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGAA
GCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACAT
TACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
S214G 463 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTggtGTTTACCCGACCCT
GAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGAG
CAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGAA
GCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACAT
TACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
S214H 464 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTcatGTTTACCCGACCCT
GAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGAG
CAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGAA
GCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACAT
TACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
S214I 465 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTattGTTTACCCGACCCT
GAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGAG
CAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGAA
GCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACAT
TACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
S214K 466 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTaaaGTTTACCCGACCCT
GAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGAG
CAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGAA
GCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACAT
TACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
S214L 467 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTctgGTTTACCCGACCCT
GAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGAG
CAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGAA
GCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACAT
TACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
S214M 468 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTatgGTTTACCCGACCCT
GAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGAG
CAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGAA
GCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACAT
TACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
S214N 469 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTaatGTTTACCCGACCCT
GAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGAG
CAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGAA
GCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACAT
TACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
S214P 470 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTccgGTTTACCCGACCCT
GAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGAG
CAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGAA
GCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACAT
TACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
S214Q 471 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTcagGTTTACCCGACCCT
GAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGAG
CAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGAA
GCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACAT
TACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
S214R 472 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTcgtGTTTACCCGACCCT
GAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGAG
CAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGAA
GCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACAT
TACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
S214S 473 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTagcGTTTACCCGACCCT
GAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGAG
CAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGAA
GCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACAT
TACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
S214T 474 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTaccGTTTACCCGACCCT
GAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGAG
CAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGAA
GCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACAT
TACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
S214V 475 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTgttGTTTACCCGACCCT
GAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGAG
CAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGAA
GCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACAT
TACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
S214W 476 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTtggGTTTACCCGACCCT
GAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGAG
CAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGAA
GCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACAT
TACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
S214Y 477 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTtatGTTTACCCGACCCT
GAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGAG
CAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGAA
GCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACAT
TACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
A53A 478 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGgcaAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGAT
CCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGACA
CGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTATG
CTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGGC
CGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAACG
CCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCGT
GAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACCC
TGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
A53C 479 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGtgtAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGATC
CTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGACAC
GCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTATGC
TTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGGCC
GAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAACGC
CAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCGTG
AGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACCCT
GAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGAG
CAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGAA
GCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACAT
TACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
A53D 480 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGgatAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGAT
CCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGACA
CGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTATG
CTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGGC
CGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAACG
CCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCGT
GAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACCC
TGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
A53E 481 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGgaaAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGAT
CCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGACA
CGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTATG
CTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGGC
CGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAACG
CCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCGT
GAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACCC
TGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
A53F 482 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGtttAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGATC
CTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGACAC
GCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTATGC
TTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGGCC
GAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAACGC
CAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCGTG
AGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACCCT
GAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGAG
CAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGAA
GCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACAT
TACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
A53G 483 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGggtAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGAT
CCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGACA
CGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTATG
CTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGGC
CGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAACG
CCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCGT
GAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACCC
TGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
A53H 484 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGcatAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGATC
CTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGACAC
GCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTATGC
TTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGGCC
GAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAACGC
CAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCGTG
AGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACCCT
GAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGAG
CAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGAA
GCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACAT
TACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
A53I 485 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGattAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGATC
CTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGACAC
GCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTATGC
TTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGGCC
GAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAACGC
CAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCGTG
AGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACCCT
GAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGAG
CAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGAA
GCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACAT
TACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
A53K 486 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGaaaAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGAT
CCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGACA
CGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTATG
CTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGGC
CGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAACG
CCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCGT
GAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACCC
TGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
A53L 487 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGctgAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGAT
CCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGACA
CGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTATG
CTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGGC
CGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAACG
CCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCGT
GAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACCC
TGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
A53M 488 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGatgAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGAT
CCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGACA
CGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTATG
CTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGGC
CGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAACG
CCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCGT
GAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACCC
TGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
A53N 489 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGaatAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGATC
CTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGACAC
GCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTATGC
TTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGGCC
GAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAACGC
CAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCGTG
AGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACCCT
GAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGAG
CAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGAA
GCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACAT
TACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
A53P 490 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGccgAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGAT
CCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGACA
CGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTATG
CTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGGC
CGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAACG
CCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCGT
GAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACCC
TGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
A53Q 491 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGcagAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGAT
CCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGACA
CGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTATG
CTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGGC
CGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAACG
CCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCGT
GAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACCC
TGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
A53R 492 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGcgtAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGAT
CCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGACA
CGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTATG
CTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGGC
CGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAACG
CCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCGT
GAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACCC
TGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
A53S 493 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGagcAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGAT
CCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGACA
CGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTATG
CTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGGC
CGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAACG
CCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCGT
GAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACCC
TGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
A53T 494 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGaccAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGAT
CCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGACA
CGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTATG
CTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGGC
CGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAACG
CCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCGT
GAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACCC
TGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
A53V 495 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGgttAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGATC
CTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGACAC
GCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTATGC
TTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGGCC
GAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAACGC
CAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCGTG
AGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACCCT
GAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGAG
CAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGAA
GCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACAT
TACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
A53W 496 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGtggAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGAT
CCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGACA
CGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTATG
CTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGGC
CGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAACG
CCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCGT
GAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACCC
TGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
A53Y 497 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGtatAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGATC
CTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGACAC
GCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTATGC
TTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGGCC
GAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAACGC
CAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCGTG
AGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACCCT
GAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGAG
CAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGAA
GCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACAT
TACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
V294N 498 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACCc
tgAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGAG
CAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGAA
GCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACAT
TACCGACAACcaaCGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
L219F_ 499 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
V294 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
N TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
TTCAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACAACcaaCGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
L219F_ 500 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
Q295 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
A TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
TTCAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACgttGCGCGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
V294N_ 501 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
Q295 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
A TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACCc
tgAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGAG
CAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGAA
GCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACAT
TACCGACAACGCGCGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
L219F 502 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
TTCAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACgttcaaCGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
V49A 503 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
CGTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
V294A 504 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGCGCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
V49A_ 505 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
V294A TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
CGTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGCGCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Q161S_ 506 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
V294 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
A TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGAGCATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGCGCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
V49A_ 507 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
Q161S TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
CGTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGAGCATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
S177Y 508 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCTACGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Y288H 509 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGCACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
A53Q_ 510 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
Y288H TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGCAGAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGCACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
S177Y_ 511 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
Y288 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
H TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCTACGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGCACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
A53Q_ 512 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
S177Y TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGCAGAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCTACGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
D166E 513 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGAGTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
A53T_ 514 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
Q295 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
W TTTTTAGCATGACCAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTTGGCGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
D166E_ 515 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
Q295 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
W TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGAGTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTTGGCGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
A53T_ 516 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
D166E TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGACCAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGAGTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
F123H 517 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTCACTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
L174V 518 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACGTTTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
S177E 519 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCGAGGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
F123H_ 520 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
S177 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
E TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTCACTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCGAGGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
L174V_ 521 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
S177 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
E TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACGTTTATTTCGAGGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
F123H_ 522 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
L174 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
V TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTCACTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACGTTTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
M106E 523 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCGAGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
A53T_ 524 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
Q161S TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGACCAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGAGCATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
M106E_ 525 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
Q161 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
S TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCGAGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGAGCATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
A53T_ 526 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
M106E TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGACCAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCGAGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Q38G 527 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTGGTGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
D166E 528 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGAGTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Q295A 529 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTGCGCGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Q38G_ 530 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
Q295A TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTGGTGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTGCGCGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
D166E_ 531 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
Q295 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
A TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGAGTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTGCGCGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Q38G_ 532 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
D166E TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTGGTGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGAGTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
K118N 533 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAACAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
S214F 534 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTTTCGTTTACCCGACCC
TGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
A53T_ 535 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
S214F TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGACCAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTTTCGTTTACCCGACCC
TGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
K118N_ 536 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
S214F TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAACAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTTTCGTTTACCCGACCC
TGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
A53T_ 537 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
K118N TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGACCAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAACAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
S177W 538 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGgctAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGAT
CCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGACA
CGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTATG
CTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGGC
CGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAACG
CCAGGTAAACCTGTATTTCTGGGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCGT
GAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACCcT
gAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGAGC
AGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGAAG
CGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACATT
ACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
L219F 539 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGgctAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGAT
CCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGACA
CGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTATG
CTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGGC
CGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAACG
CCAGGTAAACCTGTATTTCagcGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCGTG
AGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACCTTC
AACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGAGC
AGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGAAG
CGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACATT
ACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
S177W_ 540 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
L219 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
F TTTTTAGCATGgctAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGAT
CCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGACA
CGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTATG
CTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGGC
CGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAACG
CCAGGTAAACCTGTATTTCTGGGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCGT
GAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACCT
TCAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGAG
CAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGAA
GCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACAT
TACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
A53Q_ 541 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
S177W TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGCAGAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCTGGGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACCc
tgAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGAG
CAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGAA
GCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACAT
TACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
A53Q_ 542 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
L219F TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGCAGAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCagcGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCGT
GAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACCT
TCAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGAG
CAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGAA
GCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACAT
TACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
A108G 543 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGGTATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
G205M 544 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGATGCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
A108G_ 545 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
G205 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
M TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGGTATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGATGCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Q161S_ 546 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
G205 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
M TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGAGCATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGATGCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
A108G_ 547 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
Q161 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
S TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGGTATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGAGCATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
K118Q 548 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCCAGAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
M162A 549 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGGCGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Y283L 550 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACCTGAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
V271E 551 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGAGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
K119D 552 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAGACACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
L274V 553 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTGTTACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
A232S 554 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTAGCGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Q161A 555 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGGCGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
G205L 556 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGCTGCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
S214G 557 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTGGTGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Y216A 558 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTGCGCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
N173D 559 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAGACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
F123W 560 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTGGTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
K118N 561 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAACAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
R228Q 562 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCAGCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCG
AGCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAG
AAGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCAC
ATTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Q161W 563 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGTGGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
V294F 564 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACTTCCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
S214R 565 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTCGTGTTTACCCGACCC
TGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
G286E 566 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGAGGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
R228E 567 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACGAGCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCG
AGCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAG
AAGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCAC
ATTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
C230S 568 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGAGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCG
AGCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAG
AAGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCAC
ATTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
D227E 569 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGAGCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCG
AGCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAG
AAGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCAC
ATTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
M162F 570 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGTTCACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
F123A 571 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTGCGTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
A53T_ 572 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
V294A TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGaccAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGAT
CCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGACA
CGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTATG
CTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGGC
CGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAACG
CCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCGT
GAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACCC
TGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGcgCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
A53T 573 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
Q161S_ TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
V294 TTTTTAGCATGaccAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGAT
A CCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGACA
CGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTATG
CTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGGC
CGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGagcATGACCAGCATGGATTACAAGAAACGC
CAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCGTG
AGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACCCT
GAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGAG
CAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGAA
GCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACAT
TACCGACGcgCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
A53T_ 574 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
Q161S_ TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
V294 TTTTTAGCATGaccAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGAT
Nqq CCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGACA
CGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTATG
CTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGGC
CGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGagcATGACCAGCATGGATTACAAGAAACGC
CAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCGTG
AGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACCCT
GAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGAG
CAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGAA
GCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACAT
TACCGACaacCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
A53T_ 575 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
Q295A TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGaccAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGAT
CCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGACA
CGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTATG
CTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGGC
CGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAACG
CCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCGT
GAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACCC
TGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTgcACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Q161S_ 576 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
V294 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
A_Q29 TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
5A TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGagcATGACCAGCATGGATTACAAGAAACG
CCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCGT
GAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACCC
TGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGcggcaCGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
A53T_ 577 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
Q161S_ TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
Q295 TTTTTAGCATGaccAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGAT
A CCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGACA
CGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTATG
CTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGGC
CGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGagcATGACCAGCATGGATTACAAGAAACGC
CAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCGTG
AGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACCCT
GAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGAG
CAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGAA
GCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACAT
TACCGACGTTgcACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
A53T_ 578 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
V294A_ TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
Q295 TTTTTAGCATGaccAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGAT
A CCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGACA
CGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTATG
CTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGGC
CGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAACG
CCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCGT
GAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACCC
TGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGcggcACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
A53T_ 579 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
Q161S_ TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
V294 TTTTTAGCATGaccAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGAT
A_Q29 CCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGACA
5A CGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTATG
CTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGGC
CGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGagcATGACCAGCATGGATTACAAGAAACGC
CAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCGTG
AGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACCCT
GAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGAG
CAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGAA
GCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACAT
TACCGACGcggcACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
A53T_ 580 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
Q161S_ TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
V294 TTTTTAGCATGaccAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGAT
N_Q29 CCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGACA
5A CGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTATG
CTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGGC
CGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGagcATGACCAGCATGGATTACAAGAAACGC
CAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCGTG
AGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACCCT
GAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGAG
CAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGAA
GCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACAT
TACCGACaacgcACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
A53T_ 581 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
Q295 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
W TTTTTAGCATGaccAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGAT
CCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGACA
CGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTATG
CTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGGC
CGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAACG
CCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCGT
GAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACCC
TGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTtggCGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Q161S_ 582 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
V294 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
A_Q29 TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
5W TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGagcATGACCAGCATGGATTACAAGAAACG
CCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCGT
GAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACCC
TGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGcgtggCGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
A53T_ 583 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
Q161S_ TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
Q295 TTTTTAGCATGaccAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGAT
W CCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGACA
CGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTATG
CTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGGC
CGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGagcATGACCAGCATGGATTACAAGAAACGC
CAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCGTG
AGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACCCT
GAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGAG
CAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGAA
GCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACAT
TACCGACGTTtggCGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
A53T_ 584 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
V294A_ TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
Q295 TTTTTAGCATGaccAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGAT
W CCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGACA
CGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTATG
CTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGGC
CGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAACG
CCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCGT
GAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACCC
TGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGcgtggCGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
A53T_ 585 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
Q161S_ TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
V294 TTTTTAGCATGaccAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGAT
A_Q29 CCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGACA
5W CGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTATG
CTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGGC
CGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGagcATGACCAGCATGGATTACAAGAAACGC
CAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCGTG
AGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACCCT
GAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGAG
CAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGAA
GCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACAT
TACCGACGcgtggCGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
A53T_ 586 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
Q161S_ TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
V294 TTTTTAGCATGaccAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGAT
N_Q29 CCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGACA
5W CGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTATG
CTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGGC
CGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGagcATGACCAGCATGGATTACAAGAAACGC
CAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCGTG
AGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACCCT
GAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGAG
CAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGAA
GCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACAT
TACCGACaactggCGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
S177W_ 587 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
Q29S TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCtGgGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCGT
GAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACCC
TGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTgcaCGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
S177W_ 588 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
S214 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
R TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCtGgGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCGT
GAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTcgtGTTTACCCGACCCTG
AACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGAGC
AGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGAAG
CGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACATT
ACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Q161S_ 589 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
S177 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
W TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGagcATGACCAGCATGGATTACAAGAAACG
CCAGGTAAACCTGTATTTCtGgGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCGTG
AGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACCCT
GAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGAG
CAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGAA
GCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACAT
TACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
A53T_ 590 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
S177W TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGaccAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGAT
CCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGACA
CGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTATG
CTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGGC
CGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAACG
CCAGGTAAACCTGTATTTCtGgGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCGTG
AGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACCCT
GAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGAG
CAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGAA
GCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACAT
TACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
V49A_ 591 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
Q295L TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
CGTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACC
CTGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCtgCGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
V49A_ 592 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
S214R TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
CGTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTcGtGTTTACCCGACCC
TGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
A53T_ 593 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
Q295F TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGaccAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGAT
CCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGACA
CGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTATG
CTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGGC
CGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAACG
CCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCGT
GAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACCC
TGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTtttCGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
A53T_ 594 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
S214R TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGaccAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGAT
CCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGACA
CGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTATG
CTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGGC
CGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAACG
CCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCGT
GAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTcGtGTTTACCCGACCCT
GAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGAG
CAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGAA
GCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACAT
TACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
A53T_ 595 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
A161S TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
TTTTTAGCATGaccAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGAT
CCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGACA
CGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTATG
CTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGGC
CGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGagcATGACCAGCATGGATTACAAGAAACGC
CAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCGTG
AGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACCCT
GAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGAG
CAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGAA
GCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACAT
TACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Q161S_ 596 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
Q295 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
F TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGagcATGACCAGCATGGATTACAAGAAACG
CCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCGT
GAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACCC
TGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTtttCGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Q161S_ 597 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
Q295 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
L TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGagcATGACCAGCATGGATTACAAGAAACG
CCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCGT
GAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACCC
TGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCtgCGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
Q161S_ 598 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
S214 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
R TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGagcATGACCAGCATGGATTACAAGAAACG
CCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCGT
GAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTcGtGTTTACCCGACCCT
GAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGAG
CAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGAA
GCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACAT
TACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
S214R_ 599 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
Q295 TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
F TTTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGCAGATGACCAGCATGGATTACAAGAAAC
GCCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCG
TGAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTcgtGTTTACCCGACCCT
GAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGAG
CAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGAA
GCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACAT
TACCGACGTTtttCGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
V49A_ 600 ATGAGCGAAGCGGCGGACGTCGAACGTGTTTATGCAGCGATGGAAGAAGCAGCCGGTTTGCTGGGTGTGGCT
Q161S TGTGCGCGTGATAAGATTTATCCACTGCTGAGCACTTTTCAAGACACGCTGGTCGAGGGTGGCTCGGTGGTCG
CGTTTAGCATGGCTAGCGGTCGCCACAGCACCGAGCTGGACTTCTCCATCTCCGTCCCGACCAGCCACGGTGA
TCCTTACGCCACCGTTGTGGAGAAAGGTCTGTTCCCGGCGACCGGCCACCCGGTCGACGATCTGCTGGCCGAC
ACGCAAAAGCATCTGCCGGTGAGCATGTTCGCAATTGATGGTGAGGTCACGGGCGGTTTCAAAAAGACCTAT
GCTTTCTTTCCGACCGACAATATGCCTGGCGTTGCGGAGCTGAGCGCGATCCCGTCTATGCCGCCAGCGGTGG
CCGAGAATGCAGAGCTGTTCGCTCGTTACGGTCTGGATAAGGTGagcATGACCAGCATGGATTACAAGAAACG
CCAGGTAAACCTGTATTTCAGCGAGCTGTCTGCACAGACGCTCGAAGCGGAGTCCGTTTTGGCACTGGTGCGT
GAGTTGGGTCTGCACGTTCCGAATGAATTGGGTCTGAAATTCTGCAAGCGTAGCTTTAGCGTTTACCCGACCC
TGAACTGGGAAACCGGCAAGATTGACCGCCTGTGCTTTGCGGTGATCAGCAACGATCCGACGCTGGTTCCGA
GCAGCGATGAAGGCGACATCGAGAAATTTCATAACTATGCAACTAAAGCGCCGTACGCCTATGTCGGTGAGA
AGCGCACGCTGGTGTACGGTCTGACCCTGAGCCCGAAAGAAGAGTACTATAAGCTGGGCGCGTACTACCACA
TTACCGACGTTCAACGTGGCCTGCTGAAAGCGTTTGACTCTTTGGAAGATTAATGA
