CROSS-REFERENCE TO RELATED APPLICATIONS This application is a continuation of U.S. application Ser. No. 13/837,996, filed on Mar. 15, 2013, issued as U.S. Pat. No. 9,290,749 on Mar. 22, 2016, which is hereby incorporated herein by reference in its entirety for all purposes.
SEQUENCE LISTING The instant application contains a Sequence Listing which has been submitted in ASCII format via EFS-Web and is hereby incorporated by reference in its entirety. Said ASCII copy, created on Jul. 3, 2013, is named SOLAP019US_SL.txt and is 318,250 bytes in size.
BACKGROUND Certain organisms including plants and some microalgae use a type II fatty acid biosynthetic pathway, characterized by the use of discrete, monofunctional enzymes for fatty acid synthesis. In contrast, mammals and fungi use a single, large, multifunctional protein.
Type II fatty acid biosynthesis typically involves extension of a growing acyl-ACP (acyl-carrier protein) chain by two carbon units followed by cleavage by an acyl-ACP thioesterase. In plants, two main classes of acyl-ACP thioesterases have been identified: (i) those encoded by genes of the FatA class, which tend to hydrolyze oleoyl-ACP into oleate (an 18:1 fatty acid) and ACP, and (ii) those encoded by genes of the FatB class, which liberate C8-C16 fatty acids from corresponding acyl-ACP molecules.
Different FatB genes from various plants have specificities for different acyl chain lengths. As a result, different gene products will produce different fatty acid profiles in plant seeds. See, U.S. Pat. Nos. 5,850,022; 5,723,761; 5,639,790; 5,807,893; 5,455,167; 5,654,495; 5,512,482; 5,298,421; 5,667,997; and U.S. Pat. Nos. 5,344,771; 5,304,481. Recently, FatB genes have been cloned into oleaginous microalgae to produce triglycerides with altered fatty acid profiles. See, WO2010/063032, WO2011,150411, and WO2012/106560.
SUMMARY In an embodiment of the invention, there is a nucleic acid having at least 80% sequence identity to any of SEQ ID NOS: 2, 3, 5, 6, 8, 9, 11, 12, 14, 15, 17, 18, 20, 21, 23, 24, 26, 27, 29, 30, 32, 33, 35, 36, 38, 39, 41, 42, 44, 45, 47, 48, 50, 51, 53, 54, 56, 57, 59, 60, 62, 63, 65, 66, 68, 69, 71, 72, 74, 76 or any equivalent sequences by virtue of the degeneracy of the genetic code.
In another embodiment of the invention, there is a nucleic acid sequence encoding a protein having at least 80% sequence identity to any of SEQ ID NOS: 1, 4, 7, 10, 13, 16, 19, 22, 25, 28, 31, 34, 37, 40, 43, 46, 49, 52, 55, 58, 61, 64, 67, 70, 73, 75, or 77, or a fragment thereof having acyl-ACP thioesterase activity. The protein can have acyl-ACP thioesterase activity operable to alter the fatty acid profile of an oil produced by a recombinant cell comprising that sequence.
In a further embodiment of the invention there is a method of producing a recombinant host cell that produces an altered fatty acid profile, the method comprising transforming the cell with any of the nucleic acids mentioned above. The host cell can be a plant cell, a microbial cell, or a microalgal cell. Another embodiment of the invention includes a host cell produced by this method.
In an embodiment, there is a method for producing an oil or oil-derived product, the method comprising cultivating the host cell and extracting the oil, optionally wherein the cultivation is heterotrophic growth on sugar. Optionally, a fatty acid, fuel, chemical, or other oil-derived product can be produced from the oil. Optionally, the oil can have a fatty acid profile comprising at least 20% C8, C10, C12, C14 or C16 fatty acids. Optionally, the oil is produced by a microalgae and can lack C24-alpha sterols.
DESCRIPTION OF ILLUSTRATIVE EMBODIMENTS OF THE INVENTION Definitions As used with respect to nucleic acids, the term “isolated” refers to a nucleic acid that is free of at least one other component that is typically present with the naturally occurring nucleic acid. Thus, a naturally occurring nucleic acid is isolated if it has been purified away from at least one other component that occurs naturally with the nucleic acid.
A “natural oil” or “natural fat” shall mean a predominantly triglyceride oil obtained from an organism, where the oil has not undergone blending with another natural or synthetic oil, or fractionation so as to substantially alter the fatty acid profile of the triglyceride. In connection with an oil comprising triglycerides of a particular regiospecificity, the natural oil or natural fat has not been subjected to interesterification or other synthetic process to obtain that regiospecific triglyceride profile, rather the regiospecificity is produced naturally, by a cell or population of cells. In connection with a natural oil or natural fat, and as used generally throughout the present disclosure, the terms oil and fat are used interchangeably, except where otherwise noted. Thus, an “oil” or a “fat” can be liquid, solid, or partially solid at room temperature, depending on the makeup of the substance and other conditions. Here, the term “fractionation” means removing material from the oil in a way that changes its fatty acid profile relative to the profile produced by the organism, however accomplished. The terms “natural oil” and “natural fat” encompass such oils obtained from an organism, where the oil has undergone minimal processing, including refining, bleaching and/or degumming, that does not substantially change its triglyceride profile. A natural oil can also be a “noninteresterified natural oil”, which means that the natural oil has not undergone a process in which fatty acids have been redistributed in their acyl linkages to glycerol and remain essentially in the same configuration as when recovered from the organism.
“Exogenous gene” shall mean a nucleic acid that codes for the expression of an RNA and/or protein that has been introduced into a cell (e.g. by transformation/transfection), and is also referred to as a “transgene”. A cell comprising an exogenous gene may be referred to as a recombinant cell, into which additional exogenous gene(s) may be introduced. The exogenous gene may be from a different species (and so heterologous), or from the same species (and so homologous), relative to the cell being transformed. Thus, an exogenous gene can include a homologous gene that occupies a different location in the genome of the cell or is under different control, relative to the endogenous copy of the gene. An exogenous gene may be present in more than one copy in the cell. An exogenous gene may be maintained in a cell as an insertion into the genome (nuclear or plastid) or as an episomal molecule.
“Fatty acids” shall mean free fatty acids, fatty acid salts, or fatty acyl moieties in a glycerolipid. It will be understood that fatty acyl groups of glycerolipids can be described in terms of the carboxylic acid or anion of a carboxylic acid that is produced when the triglyceride is hydrolyzed or saponified.
“Microalgae” are microbial organisms that contain a chloroplast or other plastid, and optionally that are capable of performing photosynthesis, or a prokaryotic microbial organism capable of performing photosynthesis. Microalgae include obligate photoautotrophs, which cannot metabolize a fixed carbon source as energy, as well as heterotrophs, which can live solely off of a fixed carbon source. Microalgae include unicellular organisms that separate from sister cells shortly after cell division, such as Chlamydomonas, as well as microbes such as, for example, Volvox, which is a simple multicellular photosynthetic microbe of two distinct cell types. Microalgae include cells such as Chlorella, Dunaliella, and Prototheca. Microalgae also include other microbial photosynthetic organisms that exhibit cell-cell adhesion, such as Agmenellum, Anabaena, and Pyrobotrys. Microalgae also include obligate heterotrophic microorganisms that have lost the ability to perform photosynthesis, such as certain dinoflagellate algae species and species of the genus Prototheca.
An “oleaginous” cell is a cell capable of producing at least 20% lipid by dry cell weight, naturally or through recombinant or classical strain improvement. An “oleaginous microbe” or “oleaginous microorganism” is a microbe, including a microalga that is oleaginous.
In connection with a natural oil, a “profile” is the distribution of particular species or triglycerides or fatty acyl groups within the oil. A “fatty acid profile” is the distribution of fatty acyl groups in the triglycerides of the oil without reference to attachment to a glycerol backbone. Fatty acid profiles are typically determined by conversion to a fatty acid methyl ester (FAME), followed by gas chromatography (GC) analysis with flame ionization detection (FID). The fatty acid profile can be expressed as one or more percent of a fatty acid in the total fatty acid signal determined from the area under the curve for that fatty acid. FAME-GC-FID measurement approximate weight percentages of the fatty acids.
“Recombinant” is a cell, nucleic acid, protein or vector that has been modified due to the introduction of an exogenous nucleic acid or the alteration of a native nucleic acid. Thus, e.g., recombinant cells can express genes that are not found within the native (non-recombinant) form of the cell or express native genes differently than those genes are expressed by a non-recombinant cell. Recombinant cells can, without limitation, include recombinant nucleic acids that encode a gene product or suppression elements such as mutations, knockouts, antisense, interfering RNA (RNAi) or dsRNA that reduce the levels of active gene product in a cell. A “recombinant nucleic acid” is a nucleic acid originally formed in vitro, in general, by the manipulation of nucleic acid, e.g., using polymerases, ligases, exonucleases, and endonucleases, using chemical synthesis, or otherwise is in a form not normally found in nature. Recombinant nucleic acids may be produced, for example, to place two or more nucleic acids in operable linkage. Thus, an isolated nucleic acid or an expression vector formed in vitro by ligating DNA molecules that are not normally joined in nature, are both considered recombinant for the purposes of this invention. Once a recombinant nucleic acid is made and introduced into a host cell or organism, it may replicate using the in vivo cellular machinery of the host cell; however, such nucleic acids, once produced recombinantly, although subsequently replicated intracellularly, are still considered recombinant for purposes of this invention. Similarly, a “recombinant protein” is a protein made using recombinant techniques, i.e., through the expression of a recombinant nucleic acid.
Thioesterase Sequences Additional FatB genes encoding thioesterases with varying substrate preferences have been identified from plant seeds. These genes or functional subsequences thereof can be used to engineer organisms to produce fatty acids having a chain-length distribution (fatty acid profile) that is altered from the wild type organism. Specifically, recombinant cells express one or more of the exogenous FatB genes. The fatty acids can be further converted to triglycerides, fatty aldehydes, fatty alcohols and other oleochemicals either synthetically or biosynthetically. In specific embodiments, triglycerides are produced by a host cell expressing the novel FatB gene. A triglyceride-containing natural oil can be recovered from the host cell. The natural oil can be refined, degummed, bleached and/or deodorized. The oil, in its natural or processed form, can be used for foods, chemicals, fuels, cosmetics, plastics, and other uses.
The genes can be used in a variety of genetic constructs including plasmids or other vectors for expression or recombination in a host cell. The genes can be codon optimized for expression in a target host cell. The proteins produced by the genes can be used in vivo or in purified form.
The gene sequences disclosed can also be used to prepare antisense, or inhibitory RNA (e.g., RNAi or hairpin RNA) to inhibit complementary genes in a plant or other organism.
FatB genes found to be useful in producing desired fatty acid profiles in a cell are summarized below in Table 1. Nucleic acids or proteins having the sequence of SEQ ID NOS: 1-78 can be used to alter the fatty acid profile of a recombinant cell. Variant nucleic acids can also be used; e.g, variants having at least 80, 85, 90, 95, 96, 97, 98, or 99% sequence identity to SEQ ID NOS: 2, 3, 5, 6, 8, 9, 11, 12, 14, 15, 17, 18, 20, 21, 23, 24, 26, 27, 29, 30, 32, 33, 35, 36, 38, 39, 41, 42, 44, 45, 47, 48, 50, 51, 53, 54, 56, 57, 59, 60, 62, 63, 65, 66, 68, 69, 71, 72, 74, 76, or 78. Codon optimization of the genes for a variety of host organisms is contemplated, as is the use of gene fragments. Preferred codons for Prototheca strains and for Chlorella protothecoides are shown below in Tables 2 and 3, respectively. In some embodiments, the first and/or second most preferred Prototheca codons are employed for codon optimization.
In embodiments of the invention, there is protein or a nucleic acid encoding a protein having any of SEQ ID NOS: 1, 4, 7, 10, 13, 16, 19, 22, 25, 28, 31, 34, 37, 40, 43, 46, 49, 52, 55, 58, 61, 64, 67, 70, 73, 75, or 77. In an embodiment, there is protein or a nucleic acid encoding a protein having at least 70, 75, 80, 85, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% sequence identity with any of SEQ ID NOS: 1, 4, 7, 10, 13, 16, 19, 22, 25, 28, 31, 34, 37, 40, 43, 46, 49, 52, 55, 58, 61, 64, 67, 70, 73, 75, or 77. In certain embodiments, the invention encompasses a fragment any of the above-described proteins or nucleic acids (including fragments of protein or nucleic acid variants), wherein the protein fragment has acyl-ACP thioesterase activity or the nucleic acid fragment encodes such a protein fragment. In other embodiments, the fragment includes a domain of an acyl-ACP thioesterase that mediates a particular function, e.g., a specificity-determining domain. Illustrative fragments can be produced by C-terminal and/or N-terminal truncations and include at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% of the full-length sequences disclosed herein.
The term “percent sequence identity,” in the context of two or more amino acid or nucleic acid sequences, refers to two or more sequences or subsequences that are the same or have a specified percentage of amino acid residues or nucleotides that are the same, when compared and aligned for maximum correspondence, as measured using a sequence comparison algorithm or by visual inspection. For sequence comparison to determine percent nucleotide or amino acid identity, typically one sequence acts as a reference sequence, to which test sequences are compared. When using a sequence comparison algorithm, test and reference sequences are input into a computer, subsequence coordinates are designated, if necessary, and sequence algorithm program parameters are designated. The sequence comparison algorithm then calculates the percent sequence identity for the test sequence(s) relative to the reference sequence, based on the designated program parameters. Optimal alignment of sequences for comparison can be conducted using the NCBI BLAST software (ncbi.nlm.nih.gov/BLAST/) set to default parameters. For example, to compare two nucleic acid sequences, one may use blastn with the “BLAST 2 Sequences” tool Version 2.0.12 (Apr. 21, 2000) set at the following default parameters: Matrix: BLOSUM62; Reward for match: 1; Penalty for mismatch: −2; Open Gap: 5 and Extension Gap: 2 penalties; Gap x drop-off: 50; Expect: 10; Word Size: 11; Filter: on. For a pairwise comparison of two amino acid sequences, one may use the “BLAST 2 Sequences” tool Version 2.0.12 (Apr. 21, 2000) with blastp set, for example, at the following default parameters: Matrix: BLOSUM62; Open Gap: 11 and Extension Gap: 1 penalties; Gap x drop-off 50; Expect: 10; Word Size: 3; Filter: on.
In certain embodiments, percent sequence identity for variants of the nucleic acids or proteins discussed above can be calculated by using the full-length nucleic acid sequence (e.g., one of SEQ ID NOS: 2, 3, 5, 6, 8, 9, 11, 12, 14, 15, 17, 18, 20, 21, 23, 24, 26, 27, 29, 30, 32, 33, 35, 36, 38, 39, 41, 42, 44, 45, 47, 48, 50, 51, 53, 54, 56, 57, 59, 60, 62, 63, 65, 66, 68, 69, 71, 72, 74, 76, or 78) or full-length amino acid sequence (e.g., one of SEQ ID NOS: 1, 4, 7, 10, 13, 16, 19, 22, 25, 28, 31, 34, 37, 40, 43, 46, 49, 52, 55, 58, 61, 64, 67, 70, 73, 75, or 77) as the reference sequence and comparing the full-length test sequence to this reference sequence. In some embodiments relating to fragments, percent sequence identity for variants of nucleic acid or protein fragments can be calculated over the entire length of the fragment.
The nucleic acids can be in isolated form, or part of a vector or other construct, chromosome or host cell. It has been found that is many cases the full length gene (and protein) is not needed; for example, deletion of some or all of the N-terminal hydrophobic domain (typically an 18 amino acid domain starting with LPDW (SEQ ID NO: 115)) yields a still-functional gene. In addition, fusions of the specificity determining regions of the genes in Table 1 with catalytic domains of other acyl-ACP thioesterases can yield functional genes. Thus, in certain embodiments, the invention encompasses functional fragments (e.g., specificity determining regions) of the disclosed nucleic acid or amino acids fused to heterologous acyl-ACP thioesterase nucleic acid or amino acid sequences, respectively.
TABLE 1
FatB genes according to embodiments of the present invention
Native CDS Prototheca
nucloetide moriformis
Sequence Amino Acid sequence (not codon-
Variant (relative Sequence of codon- optimized
to dominant CDS (no optimized, no nucleotide
transcript additional additional sequence
Species Gene Name idenitified) tags) cloning sites) of CDS
Cinnamomum CcFATB1b M25L, M322R, SEQ ID NO: 1 SEQ ID NO: 2 SEQ ID
camphora ΔT367-D368 NO: 3
Cinnamomum CcFATB4 “wild-type” SEQ ID NO: 4 SEQ ID NO: 5 SEQ ID
camphora NO: 6
Cinnamomum CcFATB3 “wild-type” SEQ ID NO: 7 SEQ ID NO: 8 SEQ ID
camphora NO: 9
Cuphea ChsFATB1 “wild-type” SEQ ID NO: SEQ ID NO: 11 SEQ ID
hyssopifolia 10 NO: 12
Cuphea ChsFATB2 “wild-type” SEQ ID NO: SEQ ID NO: 14 SEQ ID
hyssopifolia 13 NO: 15
Cuphea ChsFATB2b +a.a.248-259 SEQ ID NO: SEQ ID NO: 17 SEQ ID
hyssopifolia 16 NO: 18
Cuphea ChsFATB3 “wild-type” SEQ ID NO: SEQ ID NO: 20 SEQ ID
hyssopifolia 19 NO: 21
Cuphea ChsFATB3b V204I, C239F, SEQ ID NO: SEQ ID NO: 23 SEQ ID
hyssopifolia E243D, M251V 22 NO: 24
Cuphea CuPSR23FATB3 “wild-type” SEQ ID NO: SEQ ID NO: 26 SEQ ID
PSR23 25 NO: 27
Cuphea CwFATB3 “wild-type” SEQ ID NO: SEQ ID NO: 29 SEQ ID
wrightii 28 NO: 30
Cuphea CwFATB4a “wild-type” SEQ ID NO: SEQ ID NO: 32 SEQ ID
wrightii 31 NO: 33
Cuphea CwFATB4b “wild-type” SEQ ID NO: SEQ ID NO: 35 SEQ ID
wrightii 34 NO: 36
Cuphea CwFATB5 “wild-type” SEQ ID NO: SEQ ID NO: 38 SEQ ID
wrightii 37 NO: 39
Cuphea ChtFATB1a “wild-type” SEQ ID NO: SEQ ID NO: 41 SEQ ID
heterophylla 40 NO: 42
Cuphea ChtFATB1b P16S, T20P, G94S, SEQ ID NO: SEQ ID NO: 44 SEQ ID
heterophylla G105W, S293F, 43 NO: 45
L305F
Cuphea ChtFATB2b “wild-type” SEQ ID NO: SEQ ID NO: 47 SEQ ID
heterophylla 46 NO: 48
Cuphea ChtFATB2a S17P, P21S, T28N, SEQ IDO NO: SEQ ID NO: 50 SEQ ID
heterophylla L30P, S33L, 49 NO: 51
G76D, S78P,
G137W
Cuphea ChtFATB2c G76D, S78P SEQ ID NO: SEQ ID NO: 53 SEQ ID
heterophylla 52 NO: 54
Cuphea ChtFATB2d S21P, T28N, SEQ ID NO: SEQ ID NO: 56 SEQ ID
heterophylla L30P, S33L, 55 NO: 57
G76D, R97L,
H124L, W127L,
I132S, K258N,
C303R, E309G,
K334T, T386A
Cuphea ChtFATB2e G76D, R97L, SEQ ID NO: SEQ ID NO: 59 SEQ ID
heterophylla H124L, I132S, 58 NO: 60
G152S, H165L,
T211N, K258N,
C303R, E309G,
K334T, T386A
Cuphea ChtFATB2f R97L, H124L, SEQ ID NO: SEQ ID NO: 62 SEQ ID
heterophylla I132S, G152S, 61 NO: 63
H165L, T211N
Cuphea ChtFATB2g A6T, A16V, S17P, SEQ ID NO: SEQ ID NO: 65 SEQ ID
heterophylla G76D, R97L, 64 NO: 66
H124L, I132S,
S143I, G152S,
A157T, H165L,
T211N, G414A
Cuphea ChtFATB3a “wild-type” SEQ ID NO: SEQ ID NO: 68 SEQ ID
heterophylla 67 NO: 69
Cuphea ChtFATB3b C67G, H72Q, SEQ ID NO: SEQ ID NO: 71 SEQ ID
heterophylla L28F, N179I 70 NO: 72
Cuphea CvisFATB1 published SEQ ID NO: N/A SEQ ID
viscosissima 73 NO: 74
Cuphea CvisFATB2 published SEQ ID NO: N/A SEQ ID
viscosissima 75 NO: 76
Cuphea CvisFATB3 published SEQ ID NO: N/A SEQ ID
viscosissima 77 NO: 78
TABLE 2
Preferred codon usage in Prototheca strains
Ala GCG 345 (0.36) ASN AAT 8 (0.04)
GCA 66 (0.07) AAC 201 (0.96)
GCT 101 (0.11)
GCC 442 (0.46)
Cys TGT 12 (0.10) Pro CCG 161 (0.29)
TGC 105 (0.90) CCA 49 (0.09)
CCT 71 (0.13)
CCC 267 (0.49)
Asp GAT 43 (0.12) Gln CAG 226 (0.82)
GAC 316 (0.88) CAA 48 (0.18)
Glu GAG 377 (0.96) Arg AGG 33 (0.06)
GAA 14 (0.04) AGA 14 (0.02)
CGG 102 (0.18)
CGA 49 (0.08)
CGT 51 (0.09)
CGC 331(0.57)
Phe TTT 89 (0.29) Ser AGT 16 (0.03)
TTC 216 (0.71) AGC 123 (0.22)
TCG 152 (0.28)
TCA 31 (0.06)
TCT 55 (0.10)
TCC 173 (0.31)
Gly GGG 92 (0.12) Thr ACG 184 (0.38)
GGA 56 (0.07) ACA 24 (0.05)
GGT 76 (0.10) ACT 21 (0.05)
GGC 559 (0.71) ACC 249 (0.52)
His CAT 42 (0.21) Val GTG 308 (0.50)
CAC 154 (0.79) GTA 9 (0.01)
GTT 35 (0.06)
GTC 262 (0.43)
Ile ATA 4 (0.01) Trp TGG 107 (1.00)
ATT 30 (0.08)
ATC 338 (0.91)
Lys AAG 284 (0.98) Tyr TAT 10 (0.05)
AAA 7 (0.02) TAC 180 (0.95)
Leu TTG 26 (0.04) Stop TGA/TAG/TAA
TTA 3 (0.00)
CTG 447 (0.61)
CTA 20 (0.03)
CTT 45 (0.06)
CTC 190 (0.26)
Met ATG 191 (1.00)
TABLE 3
Preferred codon usage in Chlorella protothecoides
TTC (Phe) TAC (Tyr) TGC (Cys) TGA (Stop)
TGG (Trp) CCC (Pro) CAC (His) CGC (Arg)
CTG (Leu) CAG (Gln) ATC (Ile) ACC (Thr)
GAC (Asp) TCC (Ser) ATG (Met) AAG (Lys)
GCC (Ala) AAC (Asn) GGC (Gly) GTG (Val)
GAG (Glu)
Host Cells The host cell can be a single cell or part of a multicellular organism such as a plant. Methods for expressing Fatb genes in a plant are given in U.S. Pat. Nos. 5,850,022; 5,723,761; 5,639,790; 5,807,893; 5,455,167; 5,654,495; 5,512,482; 5,298,421; 5,667,997; and U.S. Pat. Nos. 5,344,771; 5,304,481, or can be accomplished using other techniques generally known in plant biotechnology. Engineering of oleaginous microbes including Chlorophyta is disclosed in WO2010/063032, WO2011,150411, and WO2012/106560 and in the examples below.
Examples of oleaginous host cells include plant cells and microbial cells having a type II fatty acid biosynthetic pathway, including plastidic oleaginous cells such as those of oleaginous algae. Specific examples of microalgal cells include heterotrophic or obligate heterotrophic microalgae of the phylum Chlorophtya, the class Trebouxiophytae, the order Chlorellales, or the family Chlorellacae. Examples of oleaginous microalgae are provided in Published PCT Patent Applications WO2008/151149, WO2010/06032, WO2011/150410, and WO2011/150411, including species of Chlorella and Prototheca, a genus comprising obligate heterotrophs. The oleaginous cells can be, for example, capable of producing 25, 30, 40, 50, 60, 70, 80, 85, or about 90% oil by cell weight, ±5%. Optionally, the oils produced can be low in DHA or EPA fatty acids. For example, the oils can comprise less than 5%, 2%, or 1% DI-IA and/or EPA. The above-mentioned publications also disclose methods for cultivating such cells and extracting oil, especially from microalgal cells; such methods are applicable to the cells disclosed herein and incorporated by reference for these teachings. When microalgal cells are used they can be cultivated autotrophically (unless an obligate heterotroph) or in the dark using a sugar (e.g., glucose, fructose and/or sucrose). In any of the embodiments described herein, the cells can be heterotrophic cells comprising an exogenous invertase gene so as to allow the cells to produce oil from a sucrose feedstock. Alternately, or in addition, the cells can metabolize xylose from cellulosic feedstocks. For example, the cells can be genetically engineered to express one or more xylose metabolism genes such as those encoding an active xylose transporter, a xylulose-5-phosphate transporter, a xylose isomerase, a xylulokinase, a xylitol dehydrogenase and a xylose reductase. See WO2012/154626, “GENETICALLY ENGINEERED MICROORGANISMS THAT METABOLIZE XYLOSE”, published Nov. 15, 2012.
Oils and Related Products The oleaginous cells express one or more exogenous genes encoding fatty acid biosynthesis enzymes. As a result, some embodiments feature natural oils that were not obtainable from a non-plant or non-seed oil, or not obtainable at all.
The oleaginous cells produce a storage oil, which is primarily triacylglyceride and may be stored in storage bodies of the cell. A raw oil may be obtained from the cells by disrupting the cells and isolating the oil. WO2008/151149, WO2010/06032, WO2011/150410, and WO2011/1504 disclose heterotrophic cultivation and oil isolation techniques. For example, oil may be obtained by cultivating, drying and pressing the cells. The oils produced may be refined, bleached and deodorized (RBD) as known in the art or as described in WO2010/120939. The raw or RBD oils may be used in a variety of food, chemical, and industrial products or processes. After recovery of the oil, a valuable residual biomass remains. Uses for the residual biomass include the production of paper, plastics, absorbents, adsorbents, as animal feed, for human nutrition, or for fertilizer.
Where a fatty acid profile of a triglyceride (also referred to as a “triacylglyceride” or “TAG”) cell oil is given here, it will be understood that this refers to a nonfractionated sample of the storage oil extracted from the cell analyzed under conditions in which phospholipids have been removed or with an analysis method that is substantially insensitive to the fatty acids of the phospholipids (e.g. using chromatography and mass spectrometry). The oil may be subjected to an RBD process to remove phospholipids, free fatty acids and odors yet have only minor or negligible changes to the fatty acid profile of the triglycerides in the oil. Because the cells are oleaginous, in some cases the storage oil will constitute the bulk of all the TAGs in the cell.
The stable carbon isotope value δ13C is an expression of the ratio of 13C/12C relative to a standard (e.g. PDB, carbonite of fossil skeleton of Belemnite americana from Peedee formation of South Carolina). The stable carbon isotope value δ13C (0/00) of the oils can be related to the δ13C value of the feedstock used. In some embodiments, the oils are derived from oleaginous organisms heterotrophically grown on sugar derived from a C4 plant such as corn or sugarcane. In some embodiments the δ13C (0/00) of the oil is from −10 to −17 0/00 or from −13 to −16 0/00.
The oils produced according to the above methods in some cases are made using a microalgal host cell. As described above, the microalga can be, without limitation, fall in the classification of Chlorophyta, Trebouxiophyceae, Chlorellales, Chlorellaceae, or Chlorophyceae. It has been found that microalgae of Trebouxiophyceae can be distinguished from vegetable oils based on their sterol profiles. Oil produced by Chlorella protothecoides was found to produce sterols that appeared to be brassicasterol, ergosterol, campesterol, stigmasterol, and β-sitosterol, when detected by GC-MS. However, it is believed that all sterols produced by Chlorella have C24β stereochemistry. Thus, it is believed that the molecules detected as campesterol, stigmasterol, and β-sitosterol, are actually 22,23-dihydrobrassicasterol, proferasterol and clionasterol, respectively. Thus, the oils produced by the microalgae described above can be distinguished from plant oils by the presence of sterols with C24β stereochemistry and the absence of C24α stereochemistry in the sterols present. For example, the oils produced may contain 22,23-dihydrobrassicasterol while lacking campesterol; contain clionasterol, while lacking in β-sitosterol, and/or contain poriferasterol while lacking stigmasterol. Alternately, or in addition, the oils may contain significant amounts of Δ7-poriferasterol.
In embodiments of the present invention, oleaginous cells expressing one or more of the genes of Table 1 can produce an oil with at least 20% of C8, C10, C12, C14 or C16 fatty acids. In a specific embodiment, the level of myristate (C14:0) in the oil is greater than 30%.
Thus, in embodiments of the invention, there is a process for producing an oil, triglyceride, fatty acid, or derivative of any of these, comprising transforming a cell with any of the nucleic acids discussed herein. In another embodiment, the transformed cell is cultivated to produce an oil and, optionally, the oil is extracted. Oil extracted in this way can be used to produce food, oleochemicals or other products.
The oils discussed above alone or in combination are useful in the production of foods, fuels and chemicals (including plastics, foams, films, etc). The oils, triglycerides, fatty acids from the oils may be subjected to C—H activation, hydroamino methylation, methoxy-carbonation, ozonolysis, enzymatic transformations, epoxidation, methylation, dimerization, thiolation, metathesis, hydro-alkylation, lactonization, or other chemical processes.
After extracting the oil, a residual biomass may be left, which may have use as a fuel, as an animal feed, or as an ingredient in paper, plastic, or other product. For example, residual biomass from heterotrophic algae can be used in such products.
The described embodiments of the invention are intended to be merely exemplary and numerous variations and modifications will be apparent to those skilled in the art. All such variations and modifications are intended to be within the scope of the present invention. For example, the various triglyceride oils can be tailored in for a mixture of midchain and long chain fatty acids in order to adjust parameters such as polarity, solvency, and foam-height of the oils or chemicals made from the oils.
Example 1 Sequences of novel plant acyl-ACP thioesterases involved in seed-specific midchain (C8-C16) fatty acid biosynthesis in higher plants were isolated. Seed-specific lipid production genes were isolated through direct interrogation of RNA pools accumulating in oilseeds. Based on phylogenetic analysis, novel enzymes can be classified as members of FatB family of acyl-ACP thioesterases.
Seeds of oleaginous plants were obtained from local grocery stores or requested through USDA ARS National Plant Germplasm System (NPGS) from North Central Regional Plant Introduction Station (NCRIS) or USDA ARS North Central Soil Conservation Research Laboratory (Morris, Mich.). Dry seeds were homogenized in liquid nitrogen to powder, resuspended in cold extraction buffer containing 6-8M Urea and 3M LiCl and left on ice for a few hours to overnight at 4° C. The seed homogenate was passed through NucleoSpin Filters (Macherey-Nagel) by centrifugation at 20,000 g for 20 minutes in the refrigerated microcentrifuge (4° C.). The resulting RNA pellets were resuspended in the buffer containing 20 mM Tris HCl, pH7.5, 0.5% SDS, 100 mM NaCl, 25 mM EDTA, 2% PVPP) and RNA was subsequently extracted once with Phenol-Chloroform-Isoamyl Alcohol (25:24:1, v/v) and once with chloroform. RNA was finally precipitated with isopropyl alcohol (0.7 Vol.) in the presence of 150 mM of Na Acetate, pH5.2, washed with 80% ethanol by centrifugation, and dried. RNA samples were treated with Turbo DNAse (Lifetech) and purified further using RNeasy kits (Qiagen) following manufacturers' protocols. The resulting purified RNA samples were converted to pair-end cDNA libraries and subjected to next-generation sequencing (2×100 bp) using Illumina Hiseq 2000 platform. RNA sequence reads were assembled into corresponding seed transcriptomes using Trinity or Oases packages. Putative thioesterase-containg cDNA contigs were identified by mining transcriptomes for sequences with homology to known thioesterases. These in silico identified putative thioesterase cDNAs have been further verified by direct reverse transcription PCR analysis using seed RNA and primer pairs targeting full-length thioesterase cDNAs. The resulting amplified products were cloned and sequenced de novo to confirm authenticity of identified thioesterase genes.
To interrogate evolutionary and functional relationship between novel acyl-ACP thioesterases and the members of two existing thioesterase classes (FatA and FatB), we performed a phylogenetic analysis using published full-length (Mayer and Shanklin, 2007) and truncated (THYME database) amino acid thioesterase sequences. Novel proteins appear to group with known acyl-ACP FatB thioesterases involved in biosynthesis of C8-C16 fatty acids. Moreover, novel thioesterases appear to cluster into 3 predominant out-groups suggesting distinct functional similarity and evolutionary relatedness among members of each cluster.
The amino acid sequences of the FatB genes follow are shown in Table 4.
TABLE 4
Amino acid sequences of FatB genes
CuPSR23 FATB3 (SEQ ID NO: 25):
MVVAAATSAFFPVPAPGTSPKPGKSGNWPSSLSPTFKPKSIPNAGFQVKANASAH
PKANGSAVNLKSGSLNTQEDTSSSPPPRAFLNQLPDWSMLLTAITTVFVAAEKQWTMLD
RKSKRPDMLVDSVGLKCIVRDGLVSRQSFLIRSYEIGADRTASIETLMNHLQETSINHCK
SLGLLNDGFGRTPGMCKNDLIWVLTKMQIMVNRYPTWGDTVEINTWFSQSGKIGMASD
WLISDCNTGEILIRATSVWAMMNQKTRRFSRLPYEVRQELTPHFVDSPHVIEDNDQKLH
KFDVKTGDSIRKGLTPRWNDLDVNQHVSNVKYIGWILESMPIEVLETQELCSLTVEYRR
ECGMDSVLESVTAVDPSENGGRSQYKHLLRLEDGTDIVKSRTEWRPKNAGTNGAISTST
AKTSNGNSVS
CuPSR23 FATB3b (SEQ ID NO: 79):
MVVAAATSAFFPVPAPGTSPKPGKSGNWPSSLSPTFKPKSIPNAGFQVKANASAH
PKANGSAVNLKSGSLNTQEDTSSSPPPRAFLNQLPDWSMLLTAITTVFVAAEKQWTMLD
RKSKRPDMLVDSVGLKSIVRDGLVSRQSFLIRSYEIGADRTASIETLMNHLQETSINHCKS
LGLLNDGFGRTPGMCKNDLIWVLTKMQIIVIVNRYPTWGDTVEINTWFSQSGKIGMASD
WLISDCNTGEILIRATSVWAMMNQKTRRFSRLPYEVRQELTPHFVDSPHVIEDNDQKLH
KFDVKTGDSIRKGLTPRWNDLDVNQHVSNVKYIGWILESMPIEVLETQELCSLTVEYRR
ECGMDSVLESVTAVDPSENGGRSQYKHLLRLEDGTDIVKSRTEWRPKNAGTNGAISTST
AKTSNGNSAS
CwFATB3 (SEQ ID NO: 28):
MVVAAAASSAFFPVPAPRTTPKPGKFGNWPSSLSPPFKPKSNPNGRFQVKANVSP
HPKANGSAVSLKSGSLNTLEDPPSSPPPRTFLNQLPDWSRLRTAITTVFVAAEKQFTRLD
RKSKRPDMLVDWFGSETIVQDGLVFRERFSIRSYEIGADRTASIETLMNHLQDTSLNHCK
SVGLLNDGFGRTSEMCTRDLIWVLTKMQIVVNRYPTWGDTVEINSWFSQSGKIGMGRD
WLISDCNTGEILVRATSAWAMMNQKTRRFSKLPCEVRQEIAPHFVDAPPVIEDNDRKLH
KFDVKTGDSICKGLTPGWNDLDVNQHVSNVKYIGWILESMPTEVLETQELCSLTLEYRR
ECGRESVVESVTSMNPSKVGDRSQYQHLLRLEDGADIMKGRTEWRPKNAGTNRAIST
CwFATB3a (SEQ ID NO: 28):
MVVAAAASSAFFPVPAPRTTPKPGKFGNWPSSLSPPFKPKSNPNGRFQVKANVSP
HPKANGSAVSLKSGSLNTLEDPPSSPPPRTFLNQLPDWSRLRTAITTVFVAAEKQFTRLD
RKSKRPDMLVDWFGSETIVQDGLVFRERFSIRSYEIGADRTASIETLMNHLQDTSLNHCK
SVGLLNDGFGRTSEMCTRDLIWVLTKMQIVVNRYPTWGDTVEINSWFSQSGKIGMGRD
WLISDCNTGEILVRATSAWAMMNQKTRRFSKLPCEVRQEIAPHFVDAPPVIEDNDRKLH
KFDVKTGDSICKGLTPGWNDLDVNQHVSNVKYIGWILESMPTEVLETQELCSLTLEYRR
ECGRESVVESVTSMNPSKVGDRSQYQHLLRLEDGADIMKGRTEWRPKNAGTNRAIST
CwFATB3b (SEQ ID NO: 80):
MVVAAAASSAFFPVPAPRTTPKPGKFGNWPSSLSPPFKPKSNPNGRFQVKANVSP
HPKANGSAVSLKSGSLNTLEDLPSSPPPRTFLNQLPDWSRLRTAITTVFVAAEKQFTRLD
RKSKRPDMLVDWFGSETIVQDGLVFRERFSIRSYEIGADRTASIETLMNHLQDTSLNHCK
SVGLLNDGFGRTSEMCTRDLIWVLTKMQIVVNRYPTWGDTVEINSWFSQSGKIGMGRD
WLISDCNTGEILVRATSAWAMMNQKTRRFSKLPCEVRQEIAPHFVDAPPVIEDNDRKLH
KFDVKTGDSICKGLTPGWNDLDVNQHVSNVKYIGWILEKFWRPRSYALSPLNIGGNVE
GKVW
CwFATB3c (SEQ ID NO: 81):
MVVAAAASSAFFPVPAPRTTPKPGKFGNWPSSLSPPFKPKSNPNGRFQVKANVSP
HPKANGSAVSLKSGSLNTLEDLPSSPPPRTFLNQLPDWSRLRTAITTVFVATEKQFTRLD
RKSKRPDMLVDWFGSETIVQDGLVFRERFSIRSYEIGADRTASIETLMNHLQDTSLNHCK
SVGLLNDGFGRTSEMCTRDLIWVLTKMQIVVNRYPTWGDTVEINSWFSQSGKIGMGRD
WLISDCNTGEILVRATSAWAMMNQKTRRFSKLPCEVRQEIAPHFVDAPPVIEDNDRKLH
KFDVKTGDSICKGLTPGWNDLDVNQHVSNVKYIGWILEKFWRPRSYALSPLNIGGNVE
GKVW
CwFATB4a (SEQ ID NO: 31):
MVATAASSAFFPVPSADTSSSRPGKLGSGPSSLSPLKPKSIPNGGLQVKANASAPP
KINGSSVGLKSGGFKTQEDSPSAPPPRTFINQLPDWSMLLAAITTVFLAAEKQWMMLDW
KPKRPDMLVDPFGLGSIVQDGLVFRQNFSIRSYEIGADRTASIETVMNHLQETALNHVKI
AGLSNDGFGRTPEMYKRDLIWVVAKMQVMVNRYPTWGDTVEVNTWVAKSGKNGMR
RDWLISDCNTGEILTRASSVWVMMNQKTRRLSKIPDEVRNEIEPHFVDSAPVVEDDDRK
LPKLDENTADSIRKGLTPRWNDLDVNQHVNNVKYIGWILESTPPEVLETQELCSLTLEYR
RECGRESVLESLTAVDPSAEGYASRFQHLLRLEDGGEIVKARTEWRPKNAGINGVVPSE
ESSPGDFF
CwFATB4a.1 (SEQ ID NO: 82):
MVATAASSAFFPVPSADTSSSRPGKLGSGPSSLSPLKPKSIPNGGLQVKANASAPP
KINGSSVGLKSGGFKTQEDSPSAPPPRTFINQLPDWSMLLAAITTVFLAAEKQWMMLDW
KPKRPDMLVDPFGLGSIVQDGLVFRQNFSIRSYEIGADRTASIETVMNHLQETALNHVKI
AGLSNDGFGRTPEMYKRDLIWVVAKMQVMVNRYPTWGDTVEVNTWVAKSGKNGMR
RDWLISDCNTGEILTRASSVWVMMNQKTRRLSKIPDEVRNEIEPHFVDSAPVVEDDDRK
LPKLDENTADSIRKGLTPRWNDLDVNQHVNNVKYIGWILESTPPEVLETQELCSLTLEYR
RECGRESVLESLTAVDPSAEGYASRFQHLLRLEDGGEIVKARTEWRPKNAGINWVVPSE
ESSPGDFF
CwFATB4a.2 (SEQ ID NO: 83):
MVATAASSAFFPVPSADTSSSRPGKLGNGPSSLSPLKPKSIPNGGLQVKANASAPP
KINGSSVGLKSGSFKTQEDAPSAPPPRTFINQLPDWSMLLAAITTVFLAAEKQWMMLDW
KPKRPDMLVDPFGLGSIVQDGLVFRQNFSIRSYEIGADRTASIETVMNHLQETALNHVKI
AGLSNDGFGRTPEMYKRDLIWVVAKMQVMVNRYPTWGDTVEVNTWVAKSGKNGMR
RDWLISDCNTGEILTRASSVWVMMNQKTRRLSKIPDEVRNEIEPHFVDSAPVVEDDDRK
LPKLDENTADSIRKGLTPRWNDLDVNQHVNNVKYIGWILESTPPEVLETQELCSLTLEYR
RECGRESVLESLTAVDPSAEGYASRFQHLLRLEDGGEIVKARTEWRPKNAGINGVVPSE
ESSPGDFF
CwFATB4a.3 (SEQ ID NO: 84):
MVATAASSAFFPVPSADTSSSRPGKLGSGPSSLSPLKPKSIPNGGLQVKANASAPP
KINGSSVGLKSGGFKTQEDSPSAPPPRTFINQLPDWSMLLAAITTVFLAAEKQWMMLDW
KPKRPDMLVDPFGLGSIVQDGLVFRQNFSIRSYEIGADRTASIETVMNHLQETALNHVKI
AGLSNDGFGRTPEMYKRDLIWVVAKMQVMVNRYPTWGDTVEVNTWVAKSGKNGMR
RDWLISDCNTGEILTRASSVWVMMNQKTRRLSKIPDEVRNEIEPHFVDSAPVVEDDDRK
LPKLDENTADSIRKGLTPRWNDLDVNQHVNNVKYIGWILESTPPEVLETQELCSLTLEYR
RECGRESVLESLTAVDPSAEGYVSRFQHLLRLEDGGEIVKARTEWRPKNAGINGVVPSE
ESSPGDFF
CwFATB4b (SEQ ID NO: 34):
MVATAASSAFFPVPSADTSSSRPGKLGNGPSSLSPLKPKSIPNGGLQVKANASAPP
KINGSSVGLKSGSFKTQEDAPSAPPPRTFINQLPDWSMLLAAITTVFLAAEKQWMMLDW
KPKRPDMLVDPFGLGSIVQDGLVFRQNFSIRSYEIGADRTASIETVMNHLQETALNHVKI
AGLSSDGFGRTPAMSKRDLIWVVAKMQVMVNRYPAWGDTVEVNTWVAKSGKNGMR
RDWLISDCNTGEILTRASSVWVMMNQKTRRLSKIPDEVRNEIEPHFVDSAPVVEDDDRK
LPKLDENTADSIRKGLTPRWNDLDVNQHVNNVKYIGWILESTPAEVLETQELCSLTLEY
RRECGRESVLESLTAVDPSGEGDGSKFQHLLRLEDGGEIVKARTEWRPKNAGINGVVPS
EESSPGGDFF
CwFATB4b.1 (SEQ ID NO: 85):
MVATAASSAFFPVPSADTSSSRPGKLGSGPSSLSPLKPKSIPNGGLQVKANASAPP
KINGSSVGLKSGSFKTQEDAPSAPPPRTFrNQLPDWSMLLAAITTVFLAAEKQWMMLDW
KPKRPDMLVDPFGLGSIVQDGLVFRQNFSIRSYEIGADRTASIETVIVINHLQETALNHVKI
AGLSSDGFGRTPAMSKRDLIWVVAKMQVMVNRYPAWGDTVEVNTWVAKSGKNGMR
RDWLISDCNTGEILTRASSVWVMMNQKTRRLSKIPDEVRNEIEPHFVDSAPVVEDDDRK
LPKLDENTADSIRKGLTPRWNDLDVNQHVNNVKYIGWILESTPAEVLETQELCSLTLEY
RRECGRESVLESLTAVDPSGEGDGSKFQHLLRLEDGGEIVKARTEWRPKNAGINGVVPS
EESSPGGDFF
CwFATB5 (SEQ ID NO: 37):
MVAAAASSAFFSVPTPGTPPKPGKFGNWPSSLSVPFKPDNGGFHVKANASAHPK
ANGSAVNLKSGSLETPPRSFINQLPDLSVLLSKITTVFGAAEKQWKRPGMLVEPFGVDRI
FQDGVFFRQSFSIRSYEIGVDRTASIETLMNIFQETSLNHCKSIGLLNDGFGRTPEMCKRD
LIWVVTKIQVEVNRYPTWGDTIEVNTWVSESGKNGMGRDWLISDCRTGEILIRATSVWA
MMNQNTRRLSKFPYEVRQEIAPHFVDSAPVIEDDQKLQKLDVKTGDSIRDGLTPRWND
LDVNQHVNNVKYIGWILKSVPIEVFETQELCGVTLEYRRECGRDSVLESVTAMDPAKEG
DRCVYQHLLRLEDGADITIGRTEWRPKNAGANGAMSSGKTSNGNCLIEGRGWQPFRVV
RLIF
CwFATB5a (SEQ ID NO: 86):
MVAAAASSAFFSVPTPGTPPKPGKFGNWPSSLSVPFKPDNGGFHVKANASAHPK
ANGSAVNLKSGSLETPPRSFINQLPDLSVLLSKITTVFGAAEKQWKRPGMLVEPFGVDRI
FQDGFFFRQSFSIRSYEIGVDRTASIETLMNIFQETSLNHCKSIGLLNDGFGRTPEMCKRD
LIWVVTKIQVEVNRYPTWGDTIEVNTWVSESGKNGMGRDWLISDCRTGEILIRATSVWA
MMNQNTRRLSKFPYEVRQEIAPHFVDSAPVIEDDQKLQKLDVKTGDSIRDGLTPRWND
LDVNQHVNNVKYIGWILKSVPIEVFETQELCGVTLEYRRECGRDSVLESVTAMDPAKEG
DRCVYQHLLRLEDGADITIGRTEWRPKNAGANGAMSSGKTSNGNCLIEGRGWQPFRVV
RLIF
CwFATB5b (SEQ ID NO: 87):
MVAAAASSAFFSVPTPGTPPKPGKFGNWPSSLSVPFKPDNGGFHVKANASAHPK
ANGSAVNLKSGSLETPPRSFINQLPDLSVLLSKITTVFGAAEKQWKRPGMLVEPFGVDRI
FQDGVFFRQSFSIRSYEIGVDRTASIETLMNIFQETSLNHCKSIGLLNDGFGRTPEMCKRD
LIWVVTKIQVEVNRYPTWGDTIEVNTWVSESGKNGMGRDWLISDCRTGEILIRATSVWA
MMNQNTRRLSKFPYEVRQEIAPHFVDSAPVIEDDQKLQKLDVKTGDSIRDGLTPRWND
LDVNQHVNNVKYIGWILKSVPIEVFETQELCGVTLEYRRECGRDSVLESVTAMDPAKEG
DRCVYQHLLWLEDGADITIGRTEWRPKNAGANGAMSSGKTSNGNCLIEGRGWQPFRV
VRLIF
CwFATB5c (SEQ ID NO: 88):
MVAAAASSAFFSVPTPGTPPKPGKFGNWPSSLSVPFKPDNGGFHVKANASAHPK
ANGSAVNLKSGSLETPPRSFINQLPDLSVLLSKITTVFGAAEKQWKRPGMLVEPFGVDRI
FQDGVFFRQSFSIRSYEIGVDRTASIETLMNIFQETSLNHCKSIGLLNDGFGRTPEMCKRD
LIWVVTKIQVEVNRYPIWGDTIEVNTWVSESGKNGMGRDWLISDCRTGEILIRATSVWA
MMNQNTRRLSKFPYEVRQEIAPHFVDSAPVIEDDQKLQKLDVKTGDSIRDGLTPRWND
LDVNQHVNNVKYIGWILKSVPIEVFETQELCGVTLEYRRECGRDSVLESVTAMDPAKEG
DRCVYQHLLRLEDGADITIGRTEWRPKNAGANGAMSSGKTSNGNCLIEGMGWQPFRVV
RLIF
CwFATB5.1 (SEQ ID NO: 89):
MVAAAASSAFFSVPTPGTSPKPGKFRNWPSSLSVPFKPETNHNGGFHIKANASAH
PKANGSALNLKSGSLETQEDTSLSSPPRTFIKQLPDWSMLLSKITTVFGAAEKQLKRPGM
LVEPFGVDRIFQDGVFFRQSFSIRSYEIGADRTASIETLMNIFQETSLNHCKSIGLLNDGFG
RTPEMCKRDLIWVVTKIQVEVNRYPTWGDTIEVNTWVSESGKNGMGRDWLISDCRTGE
ILIRATSVWAMMNQNTRRLSKFPYEVRQEIAPHFVDSAPVIEDDRKLYKLNVKTGDSIR
DGLTPRWNDLDVNQHVNNVKFIGWILKSVPTKVFETQELCGVTLEYRRECGKDSVLES
VTAMDPAKEGDRSVYQHLLRLEDGADITIGRTEWRPKNAGANEAISSGKTSNGNSAS
CwFATB5.1a (SEQ ID NO: 90):
MVAAAASSAFFSVPTPGTSPKPGKFRNWPLSLSVPFKPETNHNGGFHIKANASAH
PKANGSALNLKSGSLETQEDTSLSSPPRTFIKQLPDWSMLLSKITTVFGAAEKQLKRPGM
LVEPFGVDRIFQDGVFFRQSFSIRSYEIGADRTAKETLMNIFQETSLNHCKSIGLLNDGFG
RTPEMCKRDLIWVVTKIQVEVNRYPTWGDTIEVNTWVSESGKNGMGRDWLISDCRTGE
ILIRATSVWAMMNQNTRRLSKFPYEVRQEIAPHFVDSAPVIEDDRKLYKLNVKTGDS1R
DGLTPRWNDLDVNQHVNNVKFIGWILKSVPTKVFETQELCGVTLEYRRECGKDSVLES
VTAMDPAKEGDRSVYQHLLRLEDGADITIGRTEWRPKNAGANEAISSGKTSNGNSAS
CcFATB2b (SEQ ID NO: 91):
MVTTSLASAYFSMKAVMLAPDGRGIKPRSSGLQVRAGNERNSCKVINGTKVKD
TEGLKGCSTLQGQSMLDDHFGLHGLVFRRTFAIRCYEVGPDRSTSIMAVMNHLQEAAR
NHAESLGLLGDGFGETLEMSKRDLIWVVRRTHVAVERYPAWGDTVEVEAWVGASGNT
GMRRDFLVRDCKTGHILTRCTSVSVMMNMRTRRLSKIPQEVRAEIDPLFIEKVAVKEGEI
KKLQKLNDSTADYIQGGWTPRWNDLDVNQHVNNIIYVGWIFKSVPDSISENHHLSSITLE
YRRECIRGNKLQSLTTVCGGSSEAGIICEHLLQLEDGSEVLRARTEWRPKHTDSFQGISER
FPQQEPHK
CcFATB3 (SEQ ID NO: 7):
MVATAAASAFFPVGAPATSSATSAKASMMPDNLDARGIKPKPASSSGLQVKAN
AHASPKINGSKVSTDTLKGEDTLTSSPAPRTFINQLPDWSMFLAAITTIFLAAEKQWTNL
DWKPRRPDMLADPFGIGRFMQDGLIFRQHFAIRSYEIGADRTASIETLMNHLQETALNH
VRSAGLLGDGFGATPEMSRRDLIWVVTRMQVLVDRYPAWGDIVEVETWVGASGKNG
MRRDWLVRDSQTGEILTRATSVWVMMNKRTRRLSKLPEEVRGEIGPYFIEDVAIIEEDN
RKLQKLNENTADNVRRGLTPRWSDLDVNQHVNNVKYIGWILESAPGSILESHELSCMTL
EYRRECGKDSVLQSMTAVSGGGSAAGGSPESSVECDHLLQLESGPEVVRGRTEWRPKS
ANNSRSILEMPAESL
CcFATB3b (SEQ ID NO: 92):
MVATAAASAFFPVGAPATSSATSAKASMMPDNLDARGIKPKLASSSGLQVKAN
AHASPKINGSKVSTDTLKGEDTLTSSPAPRTFINQLPDWSMFLAAITTIFLAAEKQWTNL
DWKPRRPDMLADPFGIGRFMQDGLIFRQHFAIRSYEIGADRTASIETLMNHLQETALNH
VRSAGLLGDGFGATPEMSRRDLIWVVTRMQVLVDRYPAWGDIVEVETWVGASGKNG
MRRDWLVRDSQTGEILTRATSVWVMMNKRTRRLSKLPEEVRGEIGPYFIEDVAIIEEDN
RKLQKLNENTADNVRRGLTPRWSDLDVNQHVNNVKYIGWILESAPGSILESHELSCMTL
EYRRECGKDSVLQSMTAVSGGGSAAGGSPESSVECDHLLQLESGPEVVRGRTEWRPKS
ANNSRSILEMPAESL
CcFATB3c (SEQ ID NO: 93):
MVATAAASAFFPVGAPATSSATSAKASMMPDNLDARGIKPKPASSSGLQVKAN
AHASPKINGSKVSTDTLKGEDTLTSSPAPRTFINQLPDWSMFLAAITTIFLAAEKQWTNL
DWKPRRPDMLADPFGIGRFMQDGLIFRQHFAIRSYEIGADRTASIETLMNHLQETALNH
VRSAGLLGDGFGATPEMSRRDLIWVVTRMQVLVDRYPAWGDIVEVETWVGASGKNG
MRRDWLVRDSQTGEILTRATSVWVMMNKRTRRLSKLPEEVRGEIGPYFIEDVAIIEEDN
RKLQKLNENTADNVRRGLTPRWSDLDVNQHVNNAKYIGWILESAPGSILESHELSCMTL
EYRRECGKDSVLQSMTAVSGGGSAAGGSPESSVECDHLLQLESGPEVVRGRTEWRPKS
ANNSRSILEMPAESL
ChtFATB1a (SEQ ID NO: 40):
MVAAAASSAFFSVPTPGTSTKPGNFGNWPSSLSVPFKPESNHNGGFRVKANASA
HPKANGSAVNLKSGSLETQEDTSSSSPPPRTFIKQLPDWGMLLSKITTVFGAAERQWKRP
GMLVEPFGVDRIFQDGVFFRQSFSIRSYEIGADRTASIETLMNIFQETSLNHCKSIGLLND
GFGRTPEMCKRDLIWVVTKIQVEVNRYPTWGDTIEVNTWVSESGKNGMGRDWLISDCR
TGEILIRATSVWAMMNRKTRRLSKFPYEVRQEIAPHFVDSAPVIEDDKKLHKLDVKTGD
SIRKGLTPRWNDLDVNQHVNNVKYIGWILKSVPAEVFETQELCGVTLEYRRECGRDSVL
ESVTAMDTAKEGDRSLYQHLLRLEDGADITIGRTEWRPKNAGANGAISTGKTSNENSVS
ChtFATB1a.1 (SEQ ID NO: 94):
MVAAAASSAFFSVPTPGTSPKPGNFGNWPSSLSVPFKPESNHNGGFRVKANASA
HPKANGSAVNLKSGSLETQEDTSSSSPPPRTFIKQLPDWGMLLSKITTVFGAAERQWKRP
GMLVEPFGVDRIFQDGVFFRHSFSIRSYEIGADRTASIETLMNIFQETSLNHCKSIGLLND
GFGRTPEMCKRDLIWVVTKIQVEVNRYPTWGDTIEVNTWVSESGKNGMGRDWLIGDC
RTGEILIRATSVWAMMNRKTRRLSKFPYEVRQEIAPHFVDSAPVIEDDKKLHKLDVKTG
DSIRKGLTPRWNDLDVNQHVNNVKYIGWILKSVPAEVFETQELCGVTLEYRRECGRDS
VLESVTAMDTAKEGDRSLYQHLLRLEDGADITIGRTEWRPKNAGANGALSTGKTSNGN
SVS
ChtFATB1a.2 (SEQ ID NO: 95):
MVAAAASSAFFSVPTPGTSPKPGNFGNWPSNLSVPFKPESNHNGGFRVKANASA
HPKANGSAVNLKSGSLETQEDTSSSSPPPRTFIKQLPDWGMLLSKITTVFGAAERQWKRP
GMLVEPFGVDRIFQDGVFFRQSFSIRSYEIGADRTASIETLMNIFQETSLNHCKSIGLLND
GFGRTPEMCKRDLIWVVTKIQVEVNRYPTWGDTIEVNTWVSESGKNGMGRDWLISDCR
TGEILIRATSVWAMMNRKTRRLSKFPYEVRQEIAPHFVDSAPVIEDDKKLHKLDVKTGD
SIRKGLTPRWNDFDVNQHVNNVKYIGWILKSVPAEVFETQELCGVTLEYRRECGRDSVL
ESVTAMDTAKEGDRSLYQHLLRLEDGADITIGRTEWRPKNAGANGAISTGKTSNENSVS
ChtFATB1a.3 (SEQ ID NO: 96):
MVAAAASSAFFSVPTPGTSPKPGNFGNWPSSLSVPFKPESNHNGGFRVKANASA
HPKANGSAVNLKSGSLETQEDTSSSSPPPRTFIKQLPDWGMLLSKITTVFGAAERQWKRP
GMLVEPFGVDRIFQDGVFFRQSFSIRSYEIGADRTASIETLMNIFQETSLNHCKSIGLLND
GFGRTPEMCKRDLIWVVTKIQVEVNRYPTWGDTIEVNTWVSESGKNGMGRDWLISDCR
TGEILIRATSVWAMMNRKTRRLSKFPYEVRQEIAPHFVDSAPVIEDDKKLHKLDVKTGD
SIRKGLTPRWNDFDVNQHVNNVKYIGWILKSVPAEVFETQELCGVTLEYRRECGRDSVL
ESVTAMDTAKEGDRSINQHLLRLEDGADITIGRTEWRPKNAGVNGAISTGKTSNENSVS
ChtFATB1a.4 (SEQ ID NO: 97):
MVAAAASSAFFSVPTPGTSPKPGNFGNWPSSLSVPFKPESNHNGGFRVKANASA
HPKANGSAVNLKSGSLETQEDTSSSSPPPRTFIKQLPDWSMLLSKITTVFGAAERQWKRP
GMLVEPFGVDRIFQDGVFFRQSFSIRSYEIGADRTASIETLMNIFQETSLNHCKSIGLLND
GFGRTPEMCKRDLIWVVTKIQVEVNRYPTWGDTIEVNTWVSESGKNGMGRDWLISDCR
TGEILIRATSVWAMMNRKTRRLSKFPYEVRQEIAPHFVDSAPVIEDDKKLHKLDVKTGD
SIRKGLTPRWNDFDVNQHVNNVKYIGWILKSVPAEVFETQELCGVTLEYRRECGRDSVL
ESVTAMDTAKEGDRSLYQHLLRLEDGADITIGRTEWRPKNAGANGAISTGKTSNENSVS
ChtFATB1b (SEQ ID NO: 43):
MVAAAASSAFFSVPTSGTSPKPGNFGNWPSSLSVPFKPESSHNGGFQVKANASA
HPKANGSAVNLKSGSLETQEDTSSSSPPPRTFIKQLPDWSMLLSKITTVFWAAERQWKRP
GMLVEPFGVDRIFQDGVFFRQSFSIRSYEIGADRTASIETLMNIFQETSLNHCKSIGLLND
GFGRTPEMCKRDLIWVVTKIQVEVNRYPTWGDTIEVNTWVSESGKNGMGRDWLISDCR
TGEILIRATSVWAMMNRKTRRLSKFPYEVRQEIAPHFVDSAPVIEDDKKLHKLDVKTGD
FIRKGLTPRWNDFDVNQHVNNVKYIGWILKSVPAEVFETQELCGVTLEYRRECGRDSVL
ESVTAMDTAKEGDRSLYQHLLRLEDGADITIGRTEWRPKNAGANGAISTGKTSNENSVS
ChtFATB2b (SEQ ID NO: 46):
MVVAAAASSAFFPVPASGTSPKPGKFGTWLSSSSPSYKPKSNPSGGFQVKANASA
HPKANGSAVSLKSGSLNTQEGTSSSPPPRTFLNQLPDWSRLRTAITTVFVAAEKQLTMLD
RKSKKPDMHVDWFGLEIIVQDGLVFRESFSIRSYEIGADRTASIETLMNHLQDTSLNHCK
SVGLLNDGFGRTPEMCKRDLIWVLTKMQIMVNRYPTWGDTVEINSWFSQSGKIGMGRN
WLISDCNTGEILIRATSIWAMMNQKTRRFSKLPNEVRQEIAPHFVDAPPVIEDNDRKLHK
FDVKTGDSICKGLTPEWNDLDVNQHVSNVKYIGWILESMPKEVLDTQELCSLTLEYRRE
CGRDSVLESVTAMDPSKVGDRSQYQHLLRLEDGTDIMKGRTEWRPKNAGTNGAISTGK
TSNGNSVS
ChtFATB2a (SEQ ID NO: 49):
MVVAAAASSAFFPVPAPGTTSKPGKFGNWPSSLSPSFKPKSNPNGGFQVKANAS
AHPKANGSAVSLKSGSLNTKEDTPSSPPPRTFLNQLPDWSRLRTAITTVFVAAEKQLTML
DRKSKKPDMHVDWFGLEIIVQDWLVFRESFSIRSYEIGADRTASIETLMNHLQDTSLNHC
KSVGLLNDGFGRTPEMCKRDLIWVLTKMQIMVNRYPTWGDTVEINSWFSQSGKIGMGR
NWLISDCNTGEILIRATSIWAMMNQKTRRFSKLPNEVRQEIAPHFVDAPPLIEDNDRKLH
KFDVKTGDSICKGLTPEWNDLDVNQHVSNVKYIGWILESMPKEVLDTQELCSLTLEYRR
ECGRDSVLESVTAMDPSKVGDRSQYQHLLRLEDGTDIMKGRTEWRPKNAGTNGAISTG
KTSNGNSVS
ChtFATB2c (SEQ ID NO: 52):
MVVAAAASSAFFPVPASGTSPKPGKFGTWLSSSSPSYKPKSNPSGGFQVKANASA
HPKANGSAVSLKSGSLNTKEDTPSSPPPRTFLNQLPDWNRLRTAITTVFVAAEKQLTML
DRKSKKPDMHVDWFGLEIIVQDGLVFRESFSIRSYEIGADRTASIETININHLQDTSLNHC
KSVGLLNDGFGRTPEMCKRDLIWVLTKMQIMVNRYPTWGDTVEINSWFSQSGKIGMGR
NWLISDCNTGEILIRATSIWAMMNQKTRRFSKLPNEVRQEIAPHFVDAPPVIEDNDRKLH
KFDVKTGDSICKGLTPEWNDLDVNQHVSNVKYIGWILESMPKEVLDTQELCSLTLEYRR
ECGRDSVLESVTAMDPSKVGDRSQYQHLLRLEDGTDIMKGRTEWRPKNAGTNGAISTG
KTSNGNSVS
ChtFATB2d (SEQ ID NO: 55):
MVVAAAASSAFFPVPAPGTTSKPGKFGNWPSSLSPSFKPKSNPNGGFQVKANAS
AHPKANGSAVSLKSGSLNTQEDTSSSPPPRTFLNQLPDWSRLLTAISTVFVAAEKQLTML
DRKSKRPDMLVDLFGLESIVQDGLVFRESYSIRSYEIGADRTASIETLMNHLQDTSLNHC
KSVGLLNDGFGRTPEMCKRDLIWVLTKMQIMVNRYPTWGDTVEINSWFSQSGKIGMGR
NWLISDCNTGEILIRATSIWAMMNQNTRRFSKLPNEVRQEIAPHFVDAPPVIEDNDRKLH
KFDVKTGDSIRKGLTPGWNDLDVNQHVSNVKYIGWILESMPTEVLETQELCSLTLEYRR
ECGRESVLESVTAMNPSKVGDRSQYQHLLRLEDGADIMKGRTEWRPKNAGTNGAISTG
KTSNGNSVS
ChtFATB2e (SEQ ID NO: 58):
MVVAAAASSAFFPVPASGTSPKPGKFGTWLSSSSPSYKPKSNPSGGFQVKANASA
HPKANGSAVSLKSGSLNTQEDTSSSPPPQTFLNQLPDWSRLLTAISTVFVAAEKQLTMLD
RKSKRPDMLVDWFGLESIVQDGLVFRESYSIRSYEISADRTASIETVMNLLQETSLNHCK
SMGILNDGFGRTPEMCKRDLIWVLTKMQILVNRYPNWGDTVEINSWFSQSGKIGMGRN
WLISDCNTGEILIRATSIWAMMNQNTRRFSKLPNEVRQEIAPHFVDAPPVIEDNDRKLHK
FDVKTGDSIRKGLTPGWNDLDVNQHVSNVKYIGWILESMPTEVLETQELCSLTLEYRRE
CGRDSVLESVTAMNPSKVGDRSQYQHLLRLEDGADIMKGRTEWRPKNAGTNGAISTGK
TSNGNSVS
ChtFATB2f (SEQ ID NO: 61):
MVVAAAASSAFFPVPASGTSPKPGKFGTWLSSSSPSYKPKSNPSGGFQVKANASA
HPKANGSAVSLKSGSLNTQEGTSSSPPPRTFLNQLPDWSRLLTAISTVFVAAEKQLTMLD
RKSKRPDMLVDWFGLESIVQDGLVFRESYSIRSYEISADRTASIETVMNLLQETSLNHCK
SMGILNDGFGRTPEMCKRDLIWVLTKMQILVNRYPNWGDTVEINSWFSQSGKIGMGRN
WLISDCNTGEILIRATSIWAMMNQKTRRFSKLPNEVRQEIAPHFVDAPPVIEDNDRKLHK
FDVKTGDSICKGLTPEWNDLDVNQHVSNVKYIGWILESMPKEVLDTQELCSLTLEYRRE
CGRDSVLESVTAMDPSKVGDRSQYQHLLRLEDGTDIMKGRTEWRPKNAGTNGAISTGK
TSNGNSVS
ChtFATB2g (SEQ ID NO: 64):
MVVAATASSAFFPVPVPGTSPKPGKFGTWLSSSSPSYKPKSNPSGGFQVKANASA
HPKANGSAVSLKSGSLNTQEDTSSSPPPRTFLNQLPDWSRLLTAISTVFVAAEKQLTMLD
RKSKRPDMLVDWFGLESIVQDGLVFREIYSIRSYEISADRTTSIETVMNLLQETSLNHCKS
MGILNDGFGRTPEMCKRDLIWVLTKMQILVNRYPNWGDTVEINSWFSQSGKIGMGRN
WLISDCNTGEILIRATSIWAMMNQKTRRFSKLPNEVRQEIAPHFVDAPPVIEDNDRKLHK
FDVKTGDSICKGLTPEWNDLDVNQHVSNVKYIGWILESMPKEVLDTQELCSLTLEYRRE
CGRDSVLESVTAMDPSKVGDRSQYQHLLRLEDGTDIMKGRTEWRPKNAGTNGAISTGK
TSNANSVS
ChtFATB2h (SEQ ID NO: 98):
MVVAAAASSAFFPVPASGTSPKPGKFGTWLSSSSPSYKPKSNPSGGFQVKANASA
HPKANGSAVSLKSGSLNTQEGTSSSPPPRTFLNQLPDWSRLLTAISTVFVAAEKQLTMLD
RKSKRPDMLVDWFGLESIVQDGLVFRESYSIRSYEISADRTASIETVMNLLQETSLNHCK
SMGILNDGFGRTPEMCKRDLIWVLTKMQILVNRYPNWGDTVEINSWFSQSGKIGMGRN
WLISDCNTGEILIRATSIWAMMNQNTRRFSKLPNEVRQEIAPHFVDAPPVIEDNDRKLHK
FDVKTGDSIRKGLTPGWNDLDVNQHVSNVKYIGWILESIPTEVLETQELCSLTLEYRREC
GRESVLESVTAMNPSKVGDRSQYQHLLRLEDGADIMKGRTEWRPKNAGTNGAISTGKT
SNGNSVS
ChtFATB3a (SEQ ID NO: 67):
MVATAASSAFFPVPSPDTSSRPGKLGNGSSSLRPLKPKFVANAGLQVKANASAPP
KINGSSVSLKSCSLKTHEDTPSAPPPRTFINQLPDWSMLLAAITTVFLAAEKQWMMLDW
KPKRPDMLVDPFGLGRIVQDGLVFRQNFSIRSYEIGADRTASIETVMNHLQETALNHVKS
AGLLNEGFGRTPEMYKRDLIWVVAKMQVMVNRYPTWGDTVEVNTWVAKSGKNGMR
RDWLISDCNTGEILTRASSVWVMMNQKTRKLSKIPDEVRHEIEPHFVDSAPVIEDDDWK
LPKLDEKTADSIRKGLTPKWNDLDVNQHVNNVKYIGWILESTPPEVLETQELCSLTLEY
RRECGRESVLESLTAVDPSGKGFGPQFQHLLRLEDGGEIVKGRTEWRPKTAGINGTIASG
ETSPGNS
ChtFATB3b (SEQ ID NO: 70):
MVATAASSAFFPVPSPDTSSRPGKLGNGSSSLRPLKPKFVANAGLQVKANASAPP
KINGSSVSLKSGSLKTQEDTPSAPPPRTFINQLPDWSMLLAAITTVFLAAEKQWMMLDW
KPKRPDMLVDPFGFGRIVQDGLVFRQNFSIRSYEIGADRTASIETVMNHLQETALNHVKS
AGLLIEGFGRTPEMYKRDLIWVVAKMQVMVNRYPTWGDTVEVNTWVAKSGKNGMRR
DWLISDCNTGEILTRASSVWVMMNQKTRKLSKIPDEVRHEIEPHFVDSAPVIEDDDWKL
PKLDEKTADSIRKGLTPKWNDLDVNQHVNNVKYIGWILESTPPEVLETQELCSLTLEYR
RECGRESVLESLTAVDPSGKGFGPQFQHLLRLEDGGEIVKGRTEWRPKTAGINGTIASGE
TSPGNS
ChtFATB3c (SEQ ID NO: 99):
MVATAASSAFFPVPSPDTSSRPGKLGNGSSSLRPLKPKFVANAGLQVKANASAPP
KINGSSVSLKSCSLKTHEDTPSAPPPRTFINQLPDWSMLLAAITTVFLAAEKQWMMLDW
KPKRPDMLVDPFGLGRIVQDGLVFRQNFSIRSYEIGADRTASIETVMNHLQETALNHVKS
AGLLNEGFGRTPEMYKRDLIWVVAKMQVMVNRYPTWGDTVEVNTWVAKSGKNGMR
RDWLISDCNTGEILTRASSVWVMMNQKTRKLSKIPDEVRHEIEPHFVDSAPVIEDDDRK
LPKLDEKTADSIRKGLTPKWNDLDVNQHVNNVKYIGWILESTPPEVLETQELCSLTLEY
RRECGRESVLESLTAVDPSEKGFGPQFQHLLRLEDGGEIVKGRTEWRPKTAGINGAIAFG
ETSPGDS
ChtFATB3d (SEQ ID NO: 100):
MVATAASSAFFPVPSPDTSSRPGKLGNGSSSLRPLKPKFVANAGLQVKANASAPP
KINGSSVSLKSCSLKTHEDTPSAPPPRTFINQLPDWSMLLAAITTVFLAAEKQWMMLDW
KPKRPDMLVDPFGLGRIVQDGLVFRQNFSIRSYEIGADRTASIKTVMNHLQETALNHVK
SAGLLNEGFGRTPEMYKRDLIWVVAKMQVMVNRYPTWGDTVEVNTWVAKSGKNGM
RRDWLISDCNTGEILTRASSVWVMMNQKTRKLSKIPDEVRHEIEPHFVDSAPVIEDDDW
KLPKLDEKTADSIRKGLTPKWNDLDVNQHVNNVKYIGWILESTPPEVLETQELCSLTLE
YRRECGRESVLESLTAVDPSGKGFGPQFQHLLRLEDGGEIVKGRTEWRPKTAGINGTIAS
GETSPGNS
ChtFATB3e (SEQ ID NO: 101):
MVATAASSAFFPVPSPDTSSRPGKLGNGSSSLRPLKPKFVANAGLQVKANASAPP
KINGSSVSLKSGSLKTHEDTPSAPPPRTFINQLPDWSMLLAAITTVFLAAEKQWMMLDW
KPKRPDMLVDPFGLGRIVQDGLVFRQNFSIRSYEIGADRTASIETVMNHLQETALNHVKS
AGLLNEGFGRTPEMYKRDLIWVVAKMQVMVNRYPTWGDTVEVNTWVAKSGKNGMR
RDWLISDCNTGEILTRASSVWVMMNQKTRKLSKIPDEVRHEIEPHFVDSAPVIEDDDWK
LPKLDEKTADSIRKGLTPKWNDLDVNQHVNNVKYIGWILESTPPEVLETQELCSLTLEY
RRECGRESVLESLTAVDPSGKGFGPQFQHLLRLEDGGEIVKGRTEWRPKTAGINGTIASG
ETSPGNS
ChtFATB3f (SEQ ID NO: 102):
MVATAASSAFFPVPSPDTSSRLGKLGNGSSSLRPLKPKFVANAGLQVKANASAPP
KINGSSVSLKSGSLKTQEDTPSAPPPRTFINQLPDWSMLLAAITTVFLAAEKQWMMLDW
KPKRPDMPVDPFGLGRIVQDGLVFRQNFSIRSYEIGADRTASIETVMNHLQETALNHVKS
AGLLNEGFGRTPEMYKRDLIWVVAKMQVMVNRYPTWGDTVEVNTWVAKSGKNGMR
RDWLISDCNTGEILTRASSVWVMMNQKTRKLSKIPDEVRHEIEPHFVDSAPVIEDDDWK
LPKLDEKTADSIRKGLTPKWNDLDVNQHVNNVKYIGWILESTPPEVLETQELCSLTLEY
RRECGRESVLESLTAVDPSEKGFGPQFQHLLRLEDGGEIVKGRTEWRPKTAGINGTIASG
ETSPGNS
ChtFATB3g (SEQ ID NO: 103):
MVATAASSAFFPVPSPDTSSRAGKLGNGSSSLRPLKPKFVANAGLQVKANASAPP
KINGS SVS LKSGSLKTQEDTPSAPPPRTFINQLPDWSMLLAAITTVFLAAEKQWMMLDW
KPKRPDMLVDPFGLGRIVQDGLVFRQNFSIRSYEIGADRTASIETVMNHLQETALNHVKS
AGLLNEGFGRTPEMYKRDLIWVVAKMQVMVNRYPTWGDTVEVNTWVAKSGKNGMR
RDWLISDCNTGEILTRASSVWVMMNQKTRKLSKIPDEVRHEIEPHEVDSAPVIEDDDWK
LPKLDEKTADSIRKGLTPKWNDLDVNQHVNNVKYIGWILESTPPEVLETQELCSLTLEY
RRECGRESVLESLTAVDPSGKGFGPQFQHLLRLEDGGEIVKGRTEWRPKTAGINGTIASG
ETSPGNS
ChsFATB1 (SEQ ID NO: 10):
MVATNAAAFSAYTFFLTSPTHGYSSKRLADTQNGYPGTSLKSKSTPPPAAAAAR
NGALPLLASICKCPKKADGSMQLDSSLVFGFQFYIRSYEVGADQTVSIQTVLNYLQEAAI
NHVQSAGYFGDSFGATPEMTKRNLIWVITKMQVLVDRYPAWGDVVQVDTWTCSSGKN
SMQRDWFVRDLKTGDIITRASSVWVLMNRLTRKLSKIPEAVLEEAKLFVMNTAPTVDD
NRKLPKLDGSSADYVLSGLTPRWSDLDMNQHVNNVKYIAWILESVPQSIPETHKLSAIT
VEYRRECGKNSVLQSLTNVSGDGITCGNSIIECHHLLQLETGPEILLARTEWISKEPGFRG
APIQAEKVYNNK
ChsFATB2 (SEQ ID NO: 13):
MVATAASSAFFPVPSPDASSRPGKLGNGSSSLSPLKPKLMANGGLQVKANASAP
PKINGSSVGLKSGSLKTQEDTPSAPPPRTFINQLPDWSMLLAAITTVFLAAEKQWMMLD
WKPKRPDMLVDPFGLGRIVQDGLVFRQNFSIRSYEIGADRTASIETVMNHLQETALNHV
KSAGLLNDGFGRTLEMYKRDLIWVVAKMQVMVNRYPTWGDTVEVNTWVAKSGKNG
MRRDWLISDCNTGEILTRASSVWVMMNQKTRRLSKIPDEVRHEIEPHFVDSAPVIEDDD
RKLPKLDEKTADSIRKGLTPKWNDLDVNQHVNNVKYIGWILESTPPEVLETQELCSLTL
EYRRECGRESVLESLTAVDPSGKGSGSQFQHLLRLEDGGEIVKGRTEWRPKTAGINGPIA
SGETSPGDSS
ChsFatB2b (SEQ ID NO: 16):
MVATAASSAFFPVPSPDASSRPGKLGNGSSSLSPLKPKLMANGGLQVKANASAP
PKINGSSVGLKSGSLKTQEDTPSAPPPRTFINQLPDWSMLLAAITTVFLAAEKQWMMLD
WKPKRPDMLVDPFGLGRIVQDGLVFRQNFSIRSYEIGADRTASIETVMNHLQETALNHV
KSAGLLNDGFGRTLEMYKRDLIWVVAKMQVMVNRYPTWGDTVEVNTWVAKSGKNG
MRRDWLISDCNTGEILTRASSKSQIMLPLHYCSVWVMMNQKTRRLSKIPDEVRHEIEPH
FVDSAPVIEDDDRKLPKLDEKTADSIRKGLTPKWNDLDVNQHVNNVKYIGWILESTPPE
VLETQELCSLTLEYRRECGRESVLESLTAVDPSGKGSGSQFQHLLRLEDGGEIVKGRTEW
RPKTAGINGPIASGETSPGDSS
ChsFatB2c (SEQ ID NO: 104):
MVATAASSAFFPVPSPDASSRPGKLGNGSSSLSPLKPKLMANGGLQVKANASAP
PKINGSSVGLKSGSLKTQEDTPSAPPPRTFINQLPDWSMLLAAITTVFLAAEKQWMMLD
WKPKRPDMLVDPFGLGRIVQDGLVFRQNFSIRSYEIGADRTASIETVMNHLQETALNHV
KSAGLLNDGFGRTLEMYKRDLIWVVAKMQVMVNRYPTWGDTVEVNTWVAKSGKNG
MRRDWLISDCNTGEILTRASSVWVMMNQKTRRLSKIPDEVRHEIEPHFVDSAPVIEDDD
RKLPKLDEKTADSIRKGLTPKWNDLDVNQHVNNVKYIGWILESTPPEVLETQELCSLTL
EYRRECGRESVLESLTAVDPSGKGSGSQFQHLMRLEDGGEIVKGRTEWRPKTAGINGPI
ASGETSPGDSS
ChsFatB2d (SEQ ID NO: 105):
MVATAASSAFFPVPSPDASSRPGKLGNGSSSLSPLKPKLMANGGLQVKANASAP
PKINGSSVGLKSGSLKTQEDTPSAPPPRTFINQLPDWSMLLAAITTVFLAAEKQWMMLD
WKPKRPDMLVDPFGLGRIVQDGLVFRQNFSIRSYEIGADRTASIETVMNHLQETALNHV
KSAGLLNDGFGRTPEMYKRDLIWVVAKMQVMVNRYPTWGDTVEVNTWVAKSGKNG
MRRDWLISDCNTGEILTRASSVWVMMNQKTRRLSKIPDEVRHEIEPHFVDSAPVIEDDD
RKLPKLDEKTADSIRKGLTPKWNDLDVNQHVNNVKYIGWILESTPPEVLETQELCSLTL
EYRRECGRESVLESLTAVDPSGKGSGSQFQHLLRLEDGGEIVKGRTEWRPKTAGINGPIA
SGETSPGDSS
Chs FATB3 (SEQ ID NO: 19):
MVAAEASSALFSVRTPGTSPKPGKFGNWPTSLSVPFKSKSNHNGGFQVKANASA
RPKANGSAVSLKSGSLDTQEDTSSSSSPPRTFINQLPDWSMLLSAITTVFVAAEKQWTML
DRKSKRPDMLMDPFGVDRVVQDGAVFRQSFSIRSYEIGADRTASIETLMNIFQETSLNHC
KSIGLLNDGFGRTPEMCKRDLIWVVTKMHVEVNRYPTWGDTIEVNTWVSESGKTGMG
RDWLISDCHTGEILIRATSMCAMMNQKTRRFSKFPYEVRQELAPHFVDSAPVIEDYQKL
HKLDVKTGDSICNGLTPRWNDLDVNQHVNNVKYIGWILESVPTEVFETQELCGLTLEYR
RECGRDSVLESVTAMDPSKEGDRSLYQHLLRLEDGADIAKGRTKWRPKNAGTNGAIST
GKTSNGNSIS
ChsFatb3b (SEQ ID NO: 22):
MVAAEASSALFSVRTPGTSPKPGKFGNWPTSLSVPFKSKSNHNGGFQVKANASA
RPKANGSAVSLKSGSLDTQEDTSSSSSPPRTFINQLPDWSMLLSAITTVINAAEKQWTML
DRKSKRPDMLMDPFGVDRVVQDGAVFRQSFSIRSYEIGADRTASIETLMNIFQETSLNHC
KSIGLLNDGFGRTPEMCKRDLIWVVTKMHIEVNRYPTWGDTIEVNTWVSESGKTGMGR
DWLISDFHTGDILIRATSVCAMMNQKTRRFSKFPYEVRQELAPHFVDSAPVIEDYQKLH
KLDVKTGDSICNGLTPRWNDLDVNQHVNNVKYIGWILESVPTEVFETQELCGLTLEYRR
ECGRDSVLESVTAMDPSKEGDRSLYQHLLRLEDGADIAKGRTKWRPKNAGTNGAISTG
KTSNGNSIS
ChsFatB3c (SEQ ID NO: 106):
MVAAEASSALFSVRTPGTSPKPGKFGNWPTSLSVPFKSKSNHNGGFQVKANASA
RPKANGSAVSLKSGSLDTQEDTSSSSSPPRTFINQLPDWSMLLSAITTVFVAAEKQWTML
DRKSKRPDMLMDPFGVDRVVQDGAVFRQSFSIRSYEIGADRTASIETLMNIFQETSLNHC
KSIGLLNDGFGRTPEMCKRDLIWVVTKMHVEVNRYPTWGDTIEVNTWVSESGKTGMG
RDWLISDCHTGEILIRATSMCAMMNQKTRRFSKFPYEVRQELAPHFVDSAPVIEDYQKL
HKLDVKTGDSICNGLTPRWNDLDVNQHVNNVKYIGWILESVPTEVFETQELCGLTLEYR
QECGRDSVLESVTAMDPSKEGDRSLYQHLLRLEDGTDIAKGRTKWRPKNAGKTSNGNS
IS
ChsFATB3d (SEQ ID NO: 107):
MVAAEASSALFSVRTPGTSPKPGKFGNWPSSLSVPFKSKSNHNGGFQVKANASA
RPKANGSAVSLKSGSLDTQEDASSSSSPPRTFINQLPDWSMLLSAITTVFVAAEKQWTML
DRKSKRSDMLMDPFGVDRVVQDGAVFRQSFSIRSYEIGADRTASIETLMNIFQETSLNHC
KSIGLLNDGFGRTPEMCKRDLIWVVTKMHVEVNRYPTWGDTIEVNTWVSESGKTGMG
RDWLISDCHTGEILIRATSMCAMMNQKTRRFSKFPYEVRQELAPHFVDSAPVIEDYQKL
HKLDVKTGDSICNGLTPRWNDLDVNQHVNNVKYIGWILESVPTEVFETQELCGLTLEYR
RECGRDSVLESVTAMDPSKEGDRSLYQHLLRLEDGADIAKGRTKWRPKNAGTNGAIST
GKTSNGNSIS
ChsFATB3e (SEQ ID NO: 108):
MVAAEASSALFSVRTPGTSPKPGKFGNWPSSLSVPFKSKSNHNGGFQVKANASA
RPKANGSAVSLKSGSLDTQEDASSSSSPPRTFINQLPDWSMLLSAITTVFVAAEKQWTML
DRKSKRSDMLMDPFGVDRVVQDGVVFRQSFSIRSYEIGADRTASIETLMNIFQETSLNHC
KSIGLLNDGFGRTPEMCKRDLIWVVTKMHVEVNRYPTWGDTIEVNTWVSESGKTGMG
RDWLISDCHTGEILIRATSMCAMMNQKTRRFSKFPYEVRQELAPHFVDSAPVIEDYQKL
HKLDVKTGDSICNGLTPRWNDLDVNQHVNNVKYIGWILESVPTEVFETQELCGLTLEYR
RECGRDSVLESVTAMDPSKEGDRSLYQHLLRLEDGADIAKGRTKWRPKNAGTNGAIST
GKTSNGNSIS
ChsFATB3f (SEQ ID NO: 109):
MVAAEASSALFSVRTPGTSPKPGKFGNWPSSLSVPFKSKSNHNGGFQVKANASA
RPKANGSAVSLKSGSLDTQEDTSSSSSPPRTFINQLPDWSMLLSAITTVFVAAEKQWTML
DRKSKRPDMLMDPFGVDRVVQDGAVFRQSFSIRSYEIGADRTASIETLMNIFQETSLNHC
KSIGLLNDGFGRTPEMCKRDLIWVVTKMHVEVNRYPTWGDTIEVNTWVSESGKTGMG,
RDWLISDCHTGEILIRATSMCAMMNQKTRRFSKFPYEVRQELAPHFVDSAPVIEDYQKL
HKLDVKTGDSICNGLTPRWNDLDVNQHVNNVKYIGWILESVPTEVFETQELCGLTLEYR
RECGRDSVLESVTAMDPSKEGDRSLYQHLLRLEDGADIAKGRTKWRPKNAGTNGAIST
GKTSNGNSIS
ChsFATB3g (SEQ ID NO: 110):
MVAAEASSALFSVRTPGTSPKPGKFGNWPTSLSVPFKSKSNHNGGFQVKANASA
RPKANGSAVSLKSGSLDTQEDTSSSSSPPRTFINQLPDWSMLLSAITTVFVAAEKQWTML
DRKSKRPDMLMDPFGVDRVVQDGAVFRQSFSIRSYEIGADRTASIETLMNIFQETSLNHC
KSIGLLNDGFGRTPEMCKRDLIWVVTKMHIEVNRYPTWGDTIEVNTWVSESGKTGMGR
DWLISDFHTGDILIRATSVCAMMNQKTRRFSKFPYEVRQELAPHFVDSAPVIEDYQKLH
KLDVKTGDSICNGLTPRWNDLDVNQHVNNVKYIGWILESVPTEVFETQELCGLTLEYRQ
ECGRDSVLESVTAMDPSKEGDRSLYQHLLRLEDGTDIAKGRTKWRPKNAGKTSNGNSIS
ChsFATB3h (SEQ ID NO: 111):
MVAAEASSALFSVRTPGTSPKPGKFGNWPSSLSVPFKSKSNHNGGFQVKANASA
RPKANGSAVSLKSGSLDTQEDASSSSSPPRTFINQLPDWSMLLSAITTVFVAAEKQWTML
DRKSKRSDMLMDPFGVDRVVQDGVVFRQSFSIRSYEIGADRTASIETLMNIFQETSLNHC
KSIGLLNDGFGRTPEMCKRDLIWVVTKMHIEVNRYPTWGDTIEVNTWVSESGKTGMGR
DWLISDFHTGDILIRATSVCAMMNQKTRRFSKFPYEVRQELAPHFVDSAPVIEDYQKLH
KLDVKTGDSICNGLTPRWNDLDVNQHVNNVKYIGWILESVPTEVFETQELCGLTLEYRQ
ECGRDSVLESVTAMDPSKEGDRSLYQHLLRLEDGTDIAKGRTKWRPKNAGKTSNGNSIS
ChsFATB3i (SEQ ID NO: 112):
MVAAEASSALFSVRTPGTSPKPGKFGNWPTSLSVPFKSKSNHNGGFQVKANASA
RPKANGSAVSLKSGSLDTQEDTSSSSSPPRTFINQLPDWSMLLSAITTVFVAAEKQWTML
DRICSKRPDMLMDPFGVDRVVQDGAVFRQSFSIRSYEIGADRTASIETLMNIFQETSLNHC
KSIGLLNDGFGRTPEMCKRDLIWVVTKMHVEVNRYPTWGDTIEVNTWVSESGKTGMG
RDWLISDCHTGEILIRATSMCAMMNQKTRRFSKFPYEVRQELAPHFVDSAPVIEDYQKL
HKLDVKTGDSICNGLTPRWNDLDVNQHVNNVKYIGWILESVPTEVFETQELCGLTLEYR
RECGGDSVLESVTAMDPSKEGDRSLYQHLLRLEDGADIAKGRTKWRPKNAGTNGAIST
GKTSNGNSIS
ChsFATB3j (SEQ ID NO: 113):
MVAAEASSALFSVRTPGTSPKPGKFGNWPTSLSVPFKSKSNHNGGFQVKANASA
RPKANGSAVSLKSGSLDTQEDTSSSSSPPRTFINQLPDWSMLLSAITTVFVAAEKQWTML
DRKSKRPDMLMDPFGVDRVVQDGAVFRQSFSIRSYEIGADRTASIETLMNIFQETSLNHC
KSIGLLNDGFGRTPEMCKRDLIWVVTKMHIEVNRYPTWGDTIEVNTWVSESGKTGMGR
DWLISDFHTGDILIRATSVCAMMNQKTRRFSKFPYEVRQELAPHFVDSAPVIEDYQKLH
KLDVKTGDSICNGLTPRWNDLDVNQHVNNVKYIGWILESVPTEVFETQELCGLTLEYRQ
ECGRDSVLESVTAMDPSKEGDRSLYQHLLRLEDGTDIAKGRTKWRPKNAGKTSNGNSIS
Example 2 In the example below, we detail the effect of expressing plant oilseed transcriptome-derived, heterologous thioesterases in the UTEX1435 (web.biosci.utexas.edu/utex/) strain, Strain A.
As in Example 1, RNA was extracted from dried plant seeds and submitted for paired-end sequencing using the Illumina Hiseq 2000 platform. RNA sequence reads were assembled into corresponding seed transcriptomes using Trinity or Oases packages and putative thioesterase-containing cDNA contigs were identified by mining transcriptomes for sequences with homology to known thioesterases. These in silico identified putative thioesterase cDNAs were verified by direct reverse transcription PCR analysis using seed RNA and primer pairs targeting full-length thioesterase cDNAs. The resulting amplified products were cloned and sequenced de novo to confirm authenticity of identified thioesterase genes and to identify sequence variants arising from expression of different gene alleles or diversity of sequences within a population of seeds. The resulting amino acid sequences were subjected to phylogenetic analysis using published full-length (Mayer and Shanklin, 2007) and truncated (THYME database) FatB sequences. The thioesterases that clustered with acyl-ACP FatB thioesterases, which are involved in biosynthesis of C8-C16 fatty acids, were pursued.
Construction of Transforming Vectors Expressing Acyl-ACP FatB Thioesterases 27 putative acyl-ACP FatB thioesterases from the species Cinnamomum camphora, Cuphea hyssopifolia, Cuphea PSR23, Cuphea wrightii, Cuphea heterophylla, and Cuphea viscosissima were synthesized in a codon-optimized form to reflect Prototheca moriformis (UTEX 1435) codon usage. Of the 27 genes synthesized, 24 were identified by our transcriptome sequencing efforts and the 3 genes from Cuphea viscosissima, were from published sequences in GenBank.
Transgenic strains were generated via transformation of the base strain Strain A (Prototheca moriformis, derived from UTEX 1435 by classical mutation and screening for high oil production) with a construct encoding 1 of the 27 FatB thioesterases. The construct pSZ2760 encoding Cinnamomum camphora (Cc) FATB1b is shown as an example, but identical methods were used to generate each of the remaining 26 constructs encoding the different respective thioesterases. Construct pSZ2760 can be written as 6S::CrTUB2:ScSUC2:CvNR::PmAMT3:CcFATB1b:CvNR::6S. The sequence of the transforming DNA is provided in Table 5 (pSZ2760). The relevant restriction sites in the construct from 5′-3′, BspQ1, KpnI, AscI, MfeI, EcoRI, SpeI, XhoI, SacI, BspQ1, respectively, are indicated in lowercase, bold, and underlined. BspQ1 sites delimit the 5′ and 3′ ends of the transforming DNA. Bold, lowercase sequences at the 5′ and 3′ end of the construct represent genomic DNA from UTEX 1435 that target integration to the 6S locus via homologous recombination. Proceeding in the 5′ to 3′ direction, the selection cassette has the C. reinhardtii β-tubulin promoter driving expression of the S. cerevisiae gene SUC2 (conferring the ability to grow on sucrose) and the Chlorella vulgaris Nitrate Reductase (NR) gene 3′ UTR. The promoter is indicated by lowercase, boxed text. The initiator ATG and terminator TGA for ScSUC2 are indicated by bold, uppercase italics, while the coding region is indicated with lowercase italics. The 3′ UTR is indicated by lowercase underlined text. The spacer region between the two cassettes is indicated by upper case text. The second cassette containing the codon optimized CcFATB1b gene (Table 5; pSZ2760) from Cinnamomum camphora is driven by the Prototheca moriformis endogenous AMT3 promoter, and has the Chlorella vulgaris Nitrate Reductase (NR) gene 3′ UTR. In this cassette, the AMT3 promoter is indicated by lowercase, boxed text. The initiator ATG and terminator TGA for the CcFATB1b gene are indicated in bold, uppercase italics, while the coding region is indicated by lowercase italics and the spacer region is indicated by upper case text. The 3′ UTR is indicated by lowercase underlined text. The final construct was sequenced to ensure correct reading frame and targeting sequences.
TABLE 5
pSZ2760 Transforming construct (SEQ ID NO: 114)
ctgcaggccttcctgttcctgctggccggcttcgccgccaagatcagcgcctccatgacgaacgagacgtccgaccgccccctggtgca
cttcacccccaacaagggctggatgaacgaccccaacggcctgtggtacgacgagaaggacgccaagtggcacctgtacttccagt
acaacccgaacgacaccgtctgggggacgcccttgttctggggccacgccacgtccgacgacctgaccaactgggaggaccagccc
atcgccatcgccccgaagcgcaacgactccggcgccttctccggctccatggtggtggactacaacaacacctccggcttcttcaacga
caccatcgacccgcgccagcgctgcgtggccatctggacctacaacaccccggagtccgaggagcagtacatctcctacagcctgga
cggcggctacaccttcaccgagtaccagaagaaccccgtgctggccgccaactccacccagttccgcgacccgaaggtcttctggtac
gagccctcccagaagtggatcatgaccgcggccaagtcccaggactacaagatcgagatctactcctccgacgacctgaagtcctgg
aagctggagtccgcgttcgccaacgagggcttcctcggctaccagtacgagtgccccggcctgatcgaggtccccaccgagcaggac
cccagcaagtcctactgggtgatgttcatctccatcaaccccggcgccccggccggcggctccttcaaccagtacttcgtcggcagcttc
aacggcacccacttcgaggccucgacaaccagtcccgcgtggtggacttcggcaaggactactacgccctgcagaccttcttcaaca
ccgacccgacctacgggagcgccctgggcatcgcgtgggcctccaactgggagtactccgccttcgtgcccaccaacccctggcgctc
ctccatgtccctcgtgcgcaaguctccctcaacaccgagtaccaggccaacccggagacggagctgatcaacctgaaggccgagcc
gatcctgaacatcagcaacgccggcccctggagccggttcgccaccaacaccacgttgacgaaggccaacagctacaacgtcgacc
tgtccaacagcaccggcaccctggagttcgagctggtgtacgccgtcaacaccacccagacgatctccaagtccgtgttcgcggacct
ctccctctggttcaagggcctggaggaccccgaggagtacctccgcatgggcttcgaggtgtccgcgtcctccttcttcctggaccgcgg
gaacagcaaggtgaagttcgtgaaggagaacccctacttcaccaaccgcatgagcgtgaacaaccagcccttcaagagcgagaac
gacctgtcctactacaaggtgtacggcttgctggaccagaacatcctggagctgtacttcaacgacggcgacgtcgtgtccaccaacac
ctacttcatgaccaccgggaacgccctgggctccgtgaacatgacgacgggggtggacaacctgttctacatcgacaagttccaggtg
catgaaggccgtgatgctggcccgcgacggccgcggcctgaagccccgctcctccgacctgcagctgcgcgccggcaacgcccaga
cctccctgaagatgatcaacggcaccaagttctcctacaccgagtccctgaagaagctgcccgactggtccatgctgttcgccgtgatc
accaccatcttctccgccgccgagaagcagtggaccaacctggagtggaagcccaagcccaaccccccccagctgctggacgacca
cttcggcccccacggcctggtgttccgccgcaccttcgccatccgctcctacgaggtgggccccgaccgctccacctccatcgtggccgt
gatgaaccacctgcaggaggccgccctgaaccacgccaagtccgtgggcatcctgggcgacggcttcggcaccaccctggagatgt
ccaagcgcgacctgatctgggtggtgaagcgcacccacgtggccgtggagcgctaccccgcctggggcgacaccgtggaggtgga
gtgctgggtgggcgcctccggcaacaacggccgccgccacgacttcctggtgcgcgactgcaagaccggcgagatcctgacccgct
gcacctccctgtccgtgatgatgaacacccgcacccgccgcctgtccaagatccccgaggaggtgcgcggcgagatcggccccgcct
tcatcgacaacgtggccgtgaaggacgaggagatcaagaagccccagaagctgaacgactccaccgccgactacatccagggcg
gcctgaccccccgctggaacgacctggacatcaaccagcacgtgaacaacatcaagtacgtggactggatcctggagaccgtgccc
gactccatcttcgagtcccaccacatctcctccttcaccatcgagtaccgccgcgagtgcacccgcgactccgtgctgcagtccctgacc
accgtgtccggcggctcctccgaggccggcctggtgtgcgagcacctgctgcagctggagggcggctccgaggtgctgcgcgccaag
accgagtggcgccccaagctgtccttccgcggcatctccgtgatccccgccgagtcctccgtgatggactacaaggaccacgacggcg
Constructs encoding the identified heterologous FatB genes, such as CcFATB1b from pSZ2760 in Table 6, were transformed into Strain A, and selected for the ability to grow on sucrose. Transformations, cell culture, lipid production and fatty acid analysis were all carried out as previously described. After cultivating on sucrose under low nitrogen conditions to accumulate oil, fatty acid profiles were determined by FAME-GC. The top performer from each transformation, as judged by the ability to produce the highest level of midchain fatty acids, is shown in Table 4.
TABLE 6
Alteration of Fatty Acid Profiles in S3150 upon Expression of Heterologous FatB Thioesterases
FA profile of top performer from each transformation
(%; primary lipid in Strain A background)
Species Gene Name SZ Plasmid Strain C8:0 C10:0 C12:0
Cinnamomum camphora CcFATB1b pSZ2760 A; T526; D1670-13 0 0 1
Cinnamomum camphora CcFATB4 pSZ2756 A; T525; D1666-31 0 1 33
Cinnamomum camphora CcFATB3 pSZ2755 A; T525; D1665-4 0 0 0
Cuphea hyssopifolia ChsFATB1 pSZ2778 A; T535; D1689-30 0 0 0
Cuphea hyssopifolia ChsFATB2 pSZ2796 A; T537; D1700-46 0 0 0
Cuphea hyssopifolia ChsFATB2b pSZ2792 A; T537; D1696-9 0 0 0
Cuphea hyssopifolia ChsFATB3 pSZ2797 A; T537; D1701-48 0 0 8
Cuphea hyssopifolia ChsFATB3b pSZ2795 A; T537; D1699-1 0 0 7
Cuphea PSR23 CuPSR23FATB3 pSZ2793 A; T537; D1697-13 0 1 0
Cuphea wrightii CwFATB3 pSZ2751 A; T525; D1661-22 0 2 17
Cuphea wrightii CwFATB4a pSZ2752 A; T525; D1662-30 0 0 0
Cuphea wrightii CwFATB4b pSZ2753 A; T525; D1663-29 0 0 0
Cuphea wrightii CwFATB5 pSZ2754 A; T525; D1664-39 0 0 0
Cuphea heterophylla ChtFATB1a pSZ2757 A; T525; D1667-19 0 0 5
Cuphea heterophylla ChtFATB1b pSZ2773 A; T535; D1685-29 0 0 2
Cuphea heterophylla ChtFATB2b pSZ2780 A; T535; D1691-8 0 0 0
Cuphea heterophylla ChtFATB2a pSZ2774 A; T537; D1702-24 0 0 0
Cuphea heterophylla ChtFATB2c pSZ2758 A; T525; D1668-22 0 0 3
Cuphea heterophylla ChtFATB2d pSZ2759 A; T526; D1669-19 0 0 4
Cuphea heterophylla ChtFATB2e pSZ2775 A; T535; D1686-23 0 1 2
Cuphea heterophylla ChtFATB2f pSZ2777 A; T535; D1688- 33 0 0 0
Cuphea heterophylla ChtFATB2g pSZ2794 A; T537; D1698-19 0 0 0
Cuphea heterophylla ChtFATB3a pSZ2776 A; T535; D1687-23 0 0 0
Cuphea heterophylla ChtFATB3b pSZ2779 A; T535; D1690- 31 0 0 0
Cuphea viscosissima CvisFATB1 pSZ2810 A; T540; D1711-30 0 1 0
Cuphea viscosissima CvisFATB2 pSZ2817 A; T547; D1718-1 0 0 0
Cuphea viscosissima CvisFATB3 pSZ2791 A; T537; D1695-1 0 0 0
A (parent strain): 0 0 0
FA profile of top performer from each transformation
(%; primary lipid in Strain A background)
Species C14:0 C16:0 C18:0 C18:1 C18:2 C18:3α
Cinnamomum camphora 15 26 2 46 9 1
Cinnamomum camphora 4 7 2 41 10 1
Cinnamomum camphora 3 44 3 41 8 0
Cuphea hyssopifolia 2 22 4 63 8 1
Cuphea hyssopifolia 6 53 3 32 6 0
Cuphea hyssopifolia 5 26 2 56 9 1
Cuphea hyssopifolia 34 27 2 24 5 1
Cuphea hyssopifolia 29 27 1 28 6 1
Cuphea PSR23 2 24 3 61 8 1
Cuphea wrightii 9 19 2 41 8 1
Cuphea wrightii 4 48 3 36 7 1
Cuphea wrightii 5 52 3 32 6 1
Cuphea wrightii 3 27 3 57 7 1
Cuphea heterophylla 18 27 2 39 7 1
Cuphea heterophylla 7 27 3 53 8 1
Cuphea heterophylla 2 25 3 61 8 1
Cuphea heterophylla 2 27 3 59 6 0
Cuphea heterophylla 2 23 3 58 7 1
Cuphea heterophylla 4 23 3 54 9 1
Cuphea heterophylla 3 24 3 57 8 1
Cuphea heterophylla 2 28 3 57 8 1
Cuphea heterophylla 2 22 3 62 9 1
Cuphea heterophylla 5 47 4 37 7 1
Cuphea heterophylla 6 49 5 32 7 0
Cuphea viscosissima 2 24 3 60 8 0
Cuphea viscosissima 4 51 2 36 6 0
Cuphea viscosissima 8 28 2 52 8 1
2 28 3 58 7 0
Many of the acyl-ACP FatB thioesterases were found to exhibit midchain activity when expressed in Prototheca moriformis. For example, expression of CcFATB1b causes an increase in myristate levels from 2% of total fatty acids in the parent, Strain A, to ˜15% in the D1670-13 primary transformant. Other examples include CcFATB4, which exhibits an increase in laurate levels from 0% in Strain A to ˜33%, and ChsFATB3, which exhibits an increase in myristate levels to ˜34%. Although some of the acyl-ACP thioesterases did not exhibit dramatic effects on midchain levels in the current incarnation, efforts will likely develop to optimize some of these constructs.
Sequences of the Heterologous Acyl-ACP Thioesterases Identified and Transformed into P. moriformis (UTEX 1435)
A complete listing of relevant sequences for the transforming constructs, such as the deduced amino acid sequence of the encoded acyl-ACP thioesterase, the native CDS coding sequence, the Prototheca moriformis codon-optimized coding sequence, and the nature of the sequence variants examined, is provided as SEQ ID NOS: 1-78.
Sequence Listing
SEQ ID NO 1:
Cinnamomum camphora (Cc) FATB1b variant M25L, M322R, AT367-D368 amino
acid sequence
MATTSLASAFCSMKAVMLARDGRGLKPRSSDLQLRAGNAQTSLKMINGTKFSYTESLKKLPDWSMLFAVI
TTIFSAAEKQWTNLEWKPKPNPPQLLDDHFGPHGLVFRRTFAIRSYEVGPDRSTSIVAVMNHLQEAALNH
AKSVGILGDGFGTTLEMSKRDLIWVVKRTHVAVERYPAWGDTVEVECWVGASGNNGRRHDFLVRDCKTGE
ILTRCTSLSVMMNTRTRRLSKIPEEVRGEIGPAFIDNVAVKDEEIKKPQKLNDSTADYIQGGLTPRWNDL
DINQHVNNIKYVDWILETVPDSIFESHHISSFTIEYRRECTRDSVLQSLTTVSGGSSEAGLVCEHLLQLE
GGSEVLRAKTEWRPKLSFRGISVIPAESSV*
SEQ ID NO 2:
Cinnamomum camphora (Cc) FATB1b variant M25L, M322R, 6T367-D368 coding
DNA sequence
TTAGCTTCTGCTTTCTGCTCGATGAAAGCTGTAATGTTGGCTCGTGATGGCAGGGGCTTGAAACCCAGGA
GCAGTGATTTGCAGCTGAGGGCGGGAAATGCACAAACCTCTTTGAAGATGATCAATGGGACCAAGTTCAG
TTACACAGAGAGCTTGAAAAAGTTGCCTGACTGGAGCATGCTCTTTGCAGTGATCACGACCATCTTTTCG
GCTGCTGAGAAGCAGTGGACCAATCTAGAGTGGAAGCCGAAGCCGAATCCACCCCAGTTGCTTGATGACC
ATTTTGGGCCGCATGGGTTAGTTTTCAGGCGCACCTTTGCCATCAGATCGTATGAGGTGGGACCTGACCG
CTCCACATCTATAGTGGCTGTTATGAATCACTTGCAGGAGGCTGCACTTAATCATGCGAAGAGTGTGGGA
ATTCTAGGAGATGGATTCGGTACGACGCTAGAGATGAGTAAGAGAGATCTGATATGGGTTGTGAAACGCA
CGCATGTTGCTGTGGAACGGTACCCTGCTTGGGGTGATACTGTTGAAGTAGAGTGCTGGGTTGGTGCATC
GGGAAATAATGGCAGGCGCCATGATTTCCTTGTCCGGGACTGCAAAACAGGCGAAATTCTTACAAGATGT
ACCAGTCTTTCGGTGATGATGAATACAAGGACAAGGAGGTTGTCCAAAATCCCTGAAGAAGTTAGAGGGG
AGATAGGGCCTGCATTCATTGATAATGTGGCTGTCAAGGACGAGGAAATTAAGAAACCACAGAAGCTCAA
TGACAGCACTGCAGATTACATCCAAGGAGGATTGACTCCTCGATGGAATGATTTGGATATCAATCAGCAC
GTTAACAACATCAAATACGTTGACTGGATTCTTGAGACTGTCCCAGACTCAATCTTTGAGAGTCATCATA
TTTCCAGCTTCACTATTGAATACAGGAGAGAGTGCACGAGGGATAGCGTGCTGCAGTCCCTGACCACTGT
CTCCGGTGGCTCGTCGGAAGCTGGGTTAGTGTGCGAGCACTTGCTCCAGCTTGAAGGTGGGTCTGAGGTA
TTGAGGGCAAAAACAGAGTGGAGGCCTAAGCTTAGTTTCAGAGGGATTAGTGTGATACCCGCAGAATCGA
GTGTCTAA
SEQ ID NO 3:
Cinnamomum camphora (Cc) FATB1b variant M25L, M322R, AT367-D368 coding
DNA sequence codon optimized for Prototheca moriformis
TTAGCTTCTGCTTTCTGCTCGATGAAAGCTGTAATGTTGGCTCGTGATGGCAGGGGCTTGAAACCCAGGA
GCAGTGATTTGCAGCTGAGGGCGGGAAATGCACAAACCTCTTTGAAGATGATCAATGGGACCAAGTTCAG
TTACACAGAGAGCTTGAAAAAGTTGCCTGACTGGAGCATGCTCTTTGCAGTGATCACGACCATCTTTTCG
GCTGCTGAGAAGCAGTGGACCAATCTAGAGTGGAAGCCGAAGCCGAATCCACCCCAGTTGCTTGATGACC
ATTTTGGGCCGCATGGGTTAGTTTTCAGGCGCACCTTTGCCATCAGATCGTATGAGGTGGGACCTGACCG
CTCCACATCTATAGTGGCTGTTATGAATCACTTGCAGGAGGCTGCACTTAATCATGCGAAGAGTGTGGGA
ATTCTAGGAGATGGATTCGGTACGACGCTAGAGATGAGTAAGAGAGATCTGATATGGGTTGTGAAACGCA
CGCATGTTGCTGTGGAACGGTACCCTGCTTGGGGTGATACTGTTGAAGTAGAGTGCTGGGTTGGTGCATC
GGGAAATAATGGCAGGCGCCATGATTTCCTTGTCCGGGACTGCAAAACAGGCGAAATTCTTACAAGATGT
ACCAGTCTTTCGGTGATGATGAATACAAGGACAAGGAGGTTGTCCAAAATCCCTGAAGAAGTTAGAGGGG
AGATAGGGCCTGCATTCATTGATAATGTGGCTGTCAAGGACGAGGAAATTAAGAAACCACAGAAGCTCAA
TGACAGCACTGCAGATTACATCCAAGGAGGATTGACTCCTCGATGGAATGATTTGGATATCAATCAGCAC
GTTAACAACATCAAATACGTTGACTGGATTCTTGAGACTGTCCCAGACTCAATCTTTGAGAGTCATCATA
TTTCCAGCTTCACTATTGAATACAGGAGAGAGTGCACGAGGGATAGCGTGCTGCAGTCCCTGACCACTGT
CTCCGGTGGCTCGTCGGAAGCTGGGTTAGTGTGCGAGCACTTGCTCCAGCTTGAAGGTGGGTCTGAGGTA
TTGAGGGCAAAAACAGAGTGGAGGCCTAAGCTTAGTTTCAGAGGGATTAGTGTGATACCCGCAGAATCGA
GTGTCTAA
SEQ ID NO: 4
Cinnamomum camphora (Cc) FATB4 amino acid sequence
MVTTSLASAYFSMKAVMLAPDGRGIKPRSSGLQVRAGNERNSCKVINGTKVKDTEGLKGCSTLQGQSMLD
DHFGLHGLVFRRTFAIRCYEVGPDRSTSIMAVMNHLQEAARNHAESLGLLGDGFGETLEMSKRDLIWVVR
RTHVAVERYPAWGDTVEVEAWVGASGNTGMRRDFLVRDCKTGHILTRCTSVSVMMNMRTRRLSKIPQEVR
AEIDPLFIEKVAVKEGEIKKLQKLNDSTADYIQGGWTPRWNDLDVNQHVNNIIYVGWIFKSVPDSISENH
HLSSITLEYRRECTRGNKLQSLTTVCGGSSEAGIICEHLLQLEDGSEVLRARTEWRPKHTDSFQGISERF
PQQEPHK
SEQ ID NO: 5
Cinnamomum camphora (Cc) FATB4 coding DNA sequence
ATGGTCACCACCTCTTTAGCTTCCGCTTACTTCTCGATGAAAGCTGTAATGTTGGCTCCTGACGGCAGGG
GCATAAAGCCCAGGAGCAGTGGTTTGCAGGTGAGGGCGGGAAATGAACGAAACTCTTGCAAGGTGATCAA
TGGGACCAAGGTCAAAGACACGGAGGGCTTGAAAGGGTGCAGCACGTTGCAAGGCCAGAGCATGCTTGAT
GACCATTTTGGTCTGCATGGGCTAGTTTTCAGGCGCACCTTTGCAATCAGATGCTATGAGGTTGGACCTG
ACCGCTCCACATCCATAATGGCTGTTATGAATCACTTGCAGGAAGCTGCACGTAATCATGCGGAGAGTCT
GGGACTTCTAGGAGATGGATTCGGTGAGACACTGGAGATGAGTAAGAGAGATCTGATATGGGTTGTGAGA
CGCACGCATGTTGCTGTGGAACGGTACCCTGCTTGGGGCGATACTGTTGAAGTCGAGGCCTGGGTGGGTG
CATCAGGTAACACTGGCATGCGCCGCGATTTCCTTGTCCGCGACTGCAAAACTGGCCACATTCTTACAAG
ATGTACCAGTGTTTCAGTGATGATGAATATGAGGACAAGGAGATTGTCCAAAATTCCCCAAGAAGTTAGA
GCGGAGATTGACCCTCTTTTCATTGAAAAGGTTGCTGTCAAGGAAGGGGAAATTAAAAAATTACAGAAGT
TGAATGATAGCACTGCAGATTACATTCAAGGGGGTTGGACTCCTCGATGGAATGATTTGGATGTCAATCA
GCACGTGAACAATATCATATACGTTGGCTGGATTTTTAAGAGCGTCCCAGACTCTATCTCTGAGAATCAT
CATCTTTCTAGCATCACTCTCGAATACAGGAGAGAGTGCACAAGGGGCAACAAGCTGCAGTCCCTGACCA
CTGTTTGTGGTGGCTCGTCGGAAGCTGGGATCATATGTGAGCACCTACTCCAGCTTGAGGATGGGTCTGA
GGTTTTGAGGGCAAGAACAGAGTGGAGGCCCAAGCACACCGATAGTTTCCAAGGCATTAGTGAGAGATTC
CCGCAGCAAGAACCGCATAAGTAA
SEQ ID NO: 6
Cinnamomum camphora (Cc) FATB4 coding DNA sequence codon optimized for
Prototheca moriformis
ATGGTGACCACCTCCCTGGCCTCCGCCTACTTCTCCATGAAGGCCGTGATGCTGGCCCCCGACGGCCGCG
GCATCAAGCCCCGCTCCTCCGGCCTGCAGGTGCGCGCCGGCAACGAGCGCAACTCCTGCAAGGTGATCAA
CGGCACCAAGGTGAAGGACACCGAGGGCCTGAAGGGCTGCTCCACCCTGCAGGGCCAGTCCATGCTGGAC
GACCACTTCGGCCTGCACGGCCTGGTGTTCCGCCGCACCTTCGCCATCCGCTGCTACGAGGTGGGCCCCG
ACCGCTCCACCTCCATCATGGCCGTGATGAACCACCTGCAGGAGGCCGCCCGCAACCACGCCGAGTCCCT
GGGCCTGCTGGGCGACGGCTTCGGCGAGACCCTGGAGATGTCCAAGCGCGACCTGATCTGGGTGGTGCGC
CGCACCCACGTGGCCGTGGAGCGCTACCCCGCCTGGGGCGACACCGTGGAGGTGGAGGCCTGGGTGGGCG
CCTCCGGCAACACCGGCATGCGCCGCGACTTCCTGGTGCGCGACTGCAAGACCGGCCACATCCTGACCCG
CTGCACCTCCGTGTCCGTGATGATGAACATGCGCACCCGCCGCCTGTCCAAGATCCCCCAGGAGGTGCGC
GCCGAGATCGACCCCCTGTTCATCGAGAAGGTGGCCGTGAAGGAGGGCGAGATCAAGAAGCTGCAGAAGC
TGAACGACTCCACCGCCGACTACATCCAGGGCGGCTGGACCCCCCGCTGGAACGACCTGGACGTGAACCA
GCACGTGAACAACATCATCTACGTGGGCTGGATCTTCAAGTCCGTGCCCGACTCCATCTCCGAGAACCAC
CACCTGTCCTCCATCACCCTGGAGTACCGCCGCGAGTGCACCCGCGGCAACAAGCTGCAGTCCCTGACCA
CCGTGTGCGGCGGCTCCTCCGAGGCCGGCATCATCTGCGAGCACCTGCTGCAGCTGGAGGACGGCTCCGA
GGTGCTGCGCGCCCGCACCGAGTGGCGCCCCAAGCACACCGACTCCTTCCAGGGCATCTCCGAGCGCTTC
CCCCAGCAGGAGCCCCACAAGTGA
SEQ ID NO: 7
Cinnamomum camphora (Cc) FATB3 amino acid sequence
MVATAAASAFFPVGAPATSSATSAKASMMPDNLDARGIKPKPASSSGLQVKANAHASPKINGSKVSTDTL
KGEDTLTSSPAPRTFINQLPDWSMFLAAITTIFLAAEKQWTNLDWKPRRPDMLADPFGIGRFMQDGLIFR
QHFAIRSYEIGADRTASIETLMNHLQETALNHVRSAGLLGDGFGATPEMSRRDLIWVVTRMQVLVDRYPA
WGDIVEVETWVGASGKNGMRRDWLVRDSQTGEILTRATSVWVMMNKRTRRLSKLPEEVRGEIGPYFIEDV
AIIEEDNRKLQKLNENTADNVRRGLTPRWSDLDVNQHVNNVKYIGWILESAPGSILESHELSCMTLEYRR
ECGKDSVLQSMTAVSGGGSAAGGSPESSVECDHLLQLESGPEVVRGRTEWRPKSANNSRSILEMPAESL
SEQ ID NO: 8
Cinnamomum camphora (Cc) FATB3 coding DNA sequence
ATGGTTGCCACCGCTGCTGCTTCTGCTTTCTTCCCGGTCGGTGCTCCGGCTACGTCATCTGCAACTTCAG
CCAAAGCGTCGATGATGCCTGATAATTTGGATGCCAGAGGCATCAAACCGAAGCCGGCTTCGTCCAGCGG
CTTGCAGGTTAAGGCAAATGCCCATGCCTCTCCCAAGATTAATGGTTCCAAGGTGAGCACGGATACCTTG
AAGGGGGAAGACACCTTAACTTCCTCGCCCGCCCCACGGACCTTTATCAACCAATTGCCTGACTGGAGCA
TGTTCCTTGCTGCCATCACAACTATTTTCTTGGCTGCCGAGAAGCAGTGGACGAATCTCGACTGGAAGCC
CAGAAGACCCGACATGCTTGCTGACCCGTTTGGCATCGGGAGGTTTATGCAGGATGGGCTGATTTTCAGG
CAGCACTTTGCAATCAGATCTTATGAGATTGGGGCTGATAGAACGGCGTCTATAGAGACTTTAATGAATC
ACTTGCAGGAGACTGCACTTAATCATGTGAGGAGTGCTGGACTCCTAGGTGATGGATTTGGTGCGACACC
TGAGATGAGTAGAAGAGATCTGATATGGGTTGTAACACGTATGCAGGTTCTTGTGGACCGCTACCCTGCT
TGGGGTGATATTGTTGAAGTAGAGACCTGGGTTGGTGCATCTGGAAAAAATGGTATGCGCCGTGATTGGC
TTGTTCGGGACAGCCAAACTGGTGAAATTCTCACACGAGCTACCAGTGTTTGGGTGATGATGAATAAACG
GACAAGGCGATTGTCCAAACTTCCTGAAGAAGTTAGAGGGGAAATAGGGCCTTATTTTATAGAAGATGTT
GCTATCATAGAGGAGGACAACAGGAAACTACAGAAGCTCAATGAAAACACTGCTGATAATGTTCGAAGGG
GTTTGACTCCTCGCTGGAGTGATCTGGATGTTAATCAGCATGTGAACAATGTCAAATACATTGGTTGGAT
TCTTGAGAGTGCACCAGGATCCATCTTGGAGAGTCATGAGCTTTCCTGCATGACCCTTGAATACAGGAGA
GAATGTGGGAAGGACAGTGTGCTGCAGTCAATGACTGCTGTCTCTGGTGGAGGCAGTGCAGCAGGTGGCT
CACCAGAATCTAGCGTTGAGTGTGACCACTTGCTCCAGCTAGAGAGTGGGCCTGAAGTTGTGAGGGGAAG
AACCGAGTGGAGGCCCAAGAGTGCTAATAACTCGAGGAGCATCCTGGAGATGCCGGCCGAGAGC
SEQ ID NO: 9
Cinnamomum camphora (Cc) FATB4 coding DNA sequence codon optimized for
Prototheca moriformis
ATGGTGGCCACCGCCGCCGCCTCCGCCTTCTTCCCCGTGGGCGCCCCCGCCACCTCCTCCGCCACCTCCG
CCAAGGCCTCCATGATGCCCGACAACCTGGACGCCCGCGGCATCAAGCCCAAGCCCGCCTCCTCCTCCGC
CCTGCAGGTGAAGGCCAACGCCCACGCCTCCCCCAAGATCAACGGCTCCAAGGTGTCCACCGACACCCTG
AAGGGCGAGGACACCCTGACCTCCTCCCCCGCCCCCCGCACCTTCATCAACCAGCTGCCCGACTGGTCCA
TGTTCCTGGCCGCCATCACCACCATCTTCCTGGCCGCCGAGAAGCAGTGGACCAACCTGGACTGGAAGCC
CCGCCGCCCCGACATGCTGGCCGACCCCTTCGGCATCGGCCGCTTCATGCAGGACGGCCTGATCTTCCGC
CAGCACTTCGCCATCCGCTCCTACGAGATCGGCGCCGACCGCACCGCCTCCATCGAGACCCTGATGAACC
ACCTGCAGGAGACCGCCCTGAACCACGTGCGCTCCGCCGGCCTGCTGGGCGACGGCTTCGGCGCCACCCC
CGAGATGTCCCGCCGCGACCTGATCTGGGTGGTGACCCGCATGCAGGTGCTGGTGGACCGCTACCCCGCC
TGGGGCGACATCGTGGAGGTGGAGACCTGGGTGGGCGCCTCCGGCAAGAACGGCATGCGCCGCGACTGGC
TGGTGCGCGACTCCCAGACCGGCGAGATCCTGACCCGCGCCACCTCCGTGTGGGTGATGATGAACAAGCG
CACCCGCCGCCTGTCCAAGCTGCCCGAGGAGGTGCGCGGCGAGATCGGCCCCTACTTCATCGAGGACGTG
GCCATCATCGAGGAGGACAACCGCAAGCTGCAGAAGCTGAACGAGAACACCGCCGACAACGTGCGCCGCG
GCCTGACCCCCCGCTGGTCCGACCTGGACGTGAACCAGCACGTGAACAACGTGAAGTACATCGGCTGGAT
CCTGGAGTCCGCCCCCGGCTCCATCCTGGAGTCCCACGAGCTGTCCTGCATGACCCTGGAGTACCGCCGC
GAGTGCGGCAAGGACTCCGTGCTGCAGTCCATGACCGCCGTGTCCGGCGGCGGCTCCGCCGCCGGCGGCT
CCCCCGAGTCCTCCGTGGAGTGCGACCACCTGCTGCAGCTGGAGTCCGGCCCCGAGGTGGTGCGCGGCCG
CACCGAGTGGCGCCCCAAGTCCGCCAACAACTCCCGCTCCATCCTGGAGATGCCCGCCGAGTCCCTGTGA
SEQ ID NO: 10
Cuphea hyssopifolia (Chs) FATB1 amino acid sequence
MVATNAAAFSAYTFFLTSPTHGYSSKRLADTQNGYPGTSLKSKSTPPPAAAAARNGALPLLASICKCPKK
ADGSMQLDSSLVFGFQFYIRSYEVGADQTVSIQTVLNYLQEAAINHVQSAGYFGDSFGATPEMTKRNLIW
VITKMQVLVDRYPAWGDVVQVDTWTCSSGKNSMQRDWFVRDLKTGDIITRASSVWVLMNRLTRKLSKIPE
AVLEEAKLFVMNTAPTVDDNRKLPKLDGSSADYVLSGLTPRWSDLDMNQHVNNVKYIAWILESVPQSIPE
THKLSAITVEYRRECGKNSVLQSLTNVSGDGITCGNSIIECHHLLQLETGPEILLARTEWISKEPGFRGA
PIQAEKVYNNK*
SEQ ID NO: 11
Cuphea hyssopifolia (Chs) FATB1 coding DNA sequence
ATGGTTGCCACTAATGCTGCTGCCTTTTCTGCTTATACTTTCTTCCTTACTTCACCAACTCATGGTTACT
CTTCCAAACGTCTCGCCGATACTCAAAATGGTTATCCGGGTACCTCCTTGAAATCGAAATCCACTCCTCC
ACCAGCTGCTGCTGCTGCTCGTAACGGTGCATTGCCACTGCTGGCCTCCATCTGCAAATGCCCCAAAAAG
GCTGATGGGAGTATGCAACTAGACAGCTCCTTGGTCTTCGGGTTTCAATTTTACATTAGATCATATGAAG
TGGGTGCGGATCAAACCGTGTCAATACAGACAGTACTCAATTACTTACAGGAGGCAGCCATCAATCATGT
TCAGAGTGCTGGCTATTTTGGTGATAGTTTTGGCGCCACCCCGGAAATGACCAAGAGGAACCTCATCTGG
GTTATCACTAAGATGCAGGTTTTGGTGGATCGCTATCCCGCTTGGGGCGATGTTGTTCAAGTTGATACAT
GGACCTGTAGTTCTGGTAAAAACAGCATGCAGCGTGATTGGTTCGTACGGGATCTCAAAACTGGAGATAT
TATAACAAGAGCCTCGAGCGTGTGGGTGCTGATGAATAGACTCACCAGAAAATTATCAAAAATTCCTGAA
GCAGTTCTGGAAGAAGCAAAACTTTTTGTGATGAACACTGCCCCCACCGTAGATGACAACAGGAAGCTAC
CAAAGCTGGATGGCAGCAGTGCTGATTATGTCCTCTCTGGCTTAACTCCTAGATGGAGCGACTTAGATAT
GAACCAGCATGTCAACAATGTGAAGTACATAGCCTGGATCCTTGAGAGTGTCCCTCAGAGCATACCGGAG
ACACACAAGCTGTCAGCGATAACCGTGGAGTACAGGAGAGAATGTGGCAAGAACAGCGTCCTCCAGTCTC
TGACCAACGTCTCCGGGGATGGAATCACATGTGGAAACAGTATTATCGAGTGCCACCATTTGCTTCAACT
TGAGACTGGCCCAGAGATTCTACTAGCGCGGACGGAGTGGATATCCAAGGAACCTGGGTTCAGGGGAGCT
CCAATCCAGGCAGAGAAAGTCTACAACAACAAATAA
SEQ ID NO: 12
Cuphea hyssopifolia (Chs) FATB1 coding DNA sequence codon optimized
for Prototheca moriformis
ATGGTGGCCACCAACGCCGCCGCCTTCTCCGCCTACACCTTCTTCCTGACCTCCCCCACCCACGGCTACT
CCTCCAAGCGCCTGGCCGACACCCAGAACGGCTACCCCGGCACCTCCCTGAAGTCCAAGTCCACCCCCCC
CCCCGCCGCCGCCGCCGCCCGCAACGGCGCCCTGCCCCTGCTGGCCTCCATCTGCAAGTGCCCCAAGAAG
GCCGACGGCTCCATGCAGCTGGACTCCTCCCTGGTGTTCGGCTTCCAGTTCTACATCCGCTCCTACGAGG
TGGGCGCCGACCAGACCGTGTCCATCCAGACCGTGCTGAACTACCTGCAGGAGGCCGCCATCAACCACGT
GCAGTCCGCCGGCTACTTCGGCGACTCCTTCGGCGCCACCCCCGAGATGACCAAGCGCAACCTGATCTGG
GTGATCACCAAGATGCAGGTGCTGGTGGACCGCTACCCCGCCTGGGGCGACGTGGTGCAGGTGGACACCT
GGACCTGCTCCTCCGGCAAGAACTCCATGCAGCGCGACTGGTTCGTGCGCGACCTGAAGACCGGCGACAT
CATCACCCGCGCCTCCTCCGTGTGGGTGCTGATGAACCGCCTGACCCGCAAGCTGTCCAAGATCCCCGAG
GCCGTGCTGGAGGAGGCCAACCTGTTCGTGATGAACACCGCCCCCACCGTGGACGACAACCGCAAGCTGC
CCAAGCTGGACGGCTCCTCCGCCGACTACGTGCTGTCCGGCCTGACCCCCCGCTGGTCCGACCTGGACAT
GAACCAGCACGTGAACAACGTGAAGTACATCGCCTGGATCCTGGAGTCCGTGCCCCAGTCCATCCCCGAG
ACCCACAAGCTGTCCGCCATCACCGTGGAGTACCGCCGCGAGTGCGGCAAGAACTCCGTGCTGCAGTCCC
TGACCAACGTGTCCGGCGACGGCATCACCTGCGGCAACTCCATCATCGAGTGCCACCACCTGCTGCAGCT
GGAGACCGGCCCCGAGATCCTGCTGGCCCGCACCGAGTGGATCTCCAAGGAGCCCGGCTTCCGCGGCGCC
CCCATCCAGGCCGAGAAGGTGTACAACAACAAGTGA
SEQ ID NO: 13
Cuphea hyssopifolia (Chs) FATB2 amino acid sequence
MVATAASSAFFPVPSPDASSRPGKLGNGSSSLSPLKPKLMANGGLQVKANASAPPKINGSSVGLKSGSLK
TQEDTPSAPPPRTFINQLPDWSMLLAAITTVFLAAEKQWMMLDWKPKRPDMLVDPFGLGRIVQDGLVFRQ
NFSIRSYEIGADRTASIETVMNHLQETALNHVKSAGLLNDGFGRTLEMYKRDLIWVVAKMQVMVNRYPTW
GDTVEVNTWVAKSGKNGMRRDWLISDCNTGEILTRASSVWVMMNQKTRRLSKIPDEVRHEIEPHFVDSAP
VIEDDDRKLPKLDEKTADSIRKGLTPKWNDLDVNQHVNNVKYIGWILESTPPEVLETQELCSLTLEYRRE
CGRESVLESLTAVDPSGKGSGSQFQHLLRLEDGGEIVKGRTEWRPKTAGINGPIASGETSPGDSS*
SEQ ID NO: 14
Cuphea hyssopifolia (Chs) FATB2 coding DNA sequence
ATGGTGGCTACCGCTGCAAGTTCAGCATTCTTCCCTGTGCCGTCCCCCGACGCCTCCTCTAGACCTGGAA
AGCTCGGCAATGGGTCATCGAGCTTGAGCCCCCTCAAGCCCAAATTGATGGCCAATGGCGGGTTGCAGGT
TAAGGCAAACGCCAGTGCCCCTCCTAAGATCAATGGTTCTTCGGTCGGTCTAAAGTCCGGCAGTCTCAAG
ACTCAGGAAGACACTCCTTCGGCGCCTCCTCCCCGGACTTTTATTAACCAGCTGCCTGATTGGAGTATGC
TTCTTGCTGCAATCACTACTGTCTTCTTGGCAGCAGAGAAGCAGTGGATGATGCTTGATTGGAAACCCAA
GAGGCCTGACATGCTTGTGGACCCGTTCGGATTGGGAAGGATTGTTCAAGATGGGCTTGTGTTCAGGCAG
AATTTTTCGATTAGGTCCTATGAAATAGGCGCTGATCGCACTGCGTCTATAGAGACGGTGATGAACCACT
TGCAGGAAACAGCTCTCAATCATGTTAAGAGTGCTGGGCTTCTTAATGACGGCTTTGGTCGTACTCTTGA
GATGTATAAAAGGGACCTTATTTGGGTTGTTGCAAAAATGCAGGTCATGGTTAACCGCTATCCTACTTGG
GGCGACACGGTTGAAGTGAATACTTGGGTTGCCAAGTCAGGGAAAAATGGTATGCGTCGTGATTGGCTCA
TAAGTGATTGCAATACAGGAGAAATTCTTACTAGAGCATCAAGTGTGTGGGTCATGATGAATCAAAAGAC
AAGAAGATTGTCAAAAATTCCAGATGAGGTTCGACATGAGATAGAGCCTCATTTCGTGGACTCTGCTCCC
GTCATTGAAGATGATGACCGGAAACTTCCCAAGCTGGATGAGAAGACTGCTGACTCCATCCGCAAGGGTC
TAACTCCGAAGTGGAATGACTTGGATGTCAATCAGCACGTCAACAACGTGAAGTACATTGGGTGGATTCT
TGAGAGTACTCCACCAGAAGTTCTGGAGACCCAGGAGTTATGTTCCCTTACCCTGGAATATAGGCGGGAA
TGCGGAAGGGAGAGCGTGCTGGAGTCCCTCACTGCTGTGGACCCCTCTGGAAAGGGCTCTGGGTCTCAGT
TCCAGCACCTTCTGCGGCTTGAGGATGGAGGTGAGATTGTGAAGGGGAGAACTGAGTGGCGACCCAAGAC
TGCAGGAATCAATGGGCCAATAGCATCCGGGGAGACCTCACCTGGAGACTCTTCTTAG
SEQ ID NO: 15
Cuphea hyssopifolia (Chs) FATB2 coding DNA sequence codon optimized
for Prototheca moriformis
ATGGTGGCCACCGCCGCCTCCTCCGCCTTCTTCCCCGTGCCCTCCCCCGACGCCTCCTCCCGCCCCGGCA
AGCTGGGCAACGGCTCCTCCTCCCTGTCCCCCCTGAAGCCCAAGCTGATGGCCAACGGCGGCCTGCAGGT
GAAGGCCAACGCCTCCGCCCCCCCCAAGATCAACGGCTCCTCCGTGGGCCTGAAGTCCGGCTCCCTGAAG
ACCCAGGAGGACACCCCCTCCGCCCCCCCCCCCCGCACCTTCATCAACCAGCTGCCCGACTGGTCCATGC
TGCTGGCCGCCATCACCACCGTGTTCCTGGCCGCCGAGAAGCAGTGGATGATGCTGGACTGGAAGCCCAA
GCGCCCCGACATGCTGGTGGACCCCTTCGGCCTGGGCCGCATCGTGCAGGACGGCCTGGTGTTCCGCCAG
AACTTCTCCATCCGCTCCTACGAGATCGGCGCCGACCGCACCGCCTCCATCGAGACCGTGATGAACCACC
TGCAGGAGACCGCCCTGAACCACGTGAAGTCCGCCGGCCTGCTGAACGACGGCTTCGGCCGCACCCTGGA
GATGTACAAGCGCGACCTGATCTGGGTGGTGGCCAAGATGCAGGTGATGGTGAACCGCTACCCCACCTGG
GGCGACACCGTGGAGGTGAACACCTGGGTGGCCAAGTCCGGCAAGAACGGCATGCGCCGCGACTGGCTGA
TCTCCGACTGCAACACCGGCGAGATCCTGACCCGCGCCTCCTCCGTGTGGGTGATGATGAACCAGAAGAC
CCGCCGCCTGTCCAAGATCCCCGACGAGGTGCGCCACGAGATCGAGCCCCACTTCGTGGACTCCGCCCCC
GTGATCGAGGACGACGACCGCAAGCTGCCCAAGCTGGACGAGAAGACCGCCGACTCCATCCGCAAGGGCC
TGACCCCCAAGTGGAACGACCTGGACGTGAACCAGCACGTGAACAACGTGAAGTACATCGGCTGGATCCT
GGAGTCCACCCCCCCCGAGGTGCTGGAGACCCAGGAGCTGTGCTCCCTGACCCTGGAGTACCGCCGCGAG
TGCGGCCGCGAGTCCGTGCTGGAGTCCCTGACCGCCGTGGACCCCTCCGGCAAGGGCTCCGGCTCCCAGT
TCCAGCACCTGCTGCGCCTGGAGGACGGCGGCGAGATCGTGAAGGGCCGCACCGAGTGGCGCCCCAAGAC
CGCCGGCATCAACGGCCCCATCGCCTCCGGCGAGACCTCCCCCGGCGACTCCTCCTGA
SEQ ID NO: 16
Cuphea hyssopifolia (Chs) FATB2b +a.a.248-259 variant amino acid
sequence
MVATAASSAFFPVPSPDASSRPGKLGNGSSSLSPLKPKLMANGGLQVKANASAPPKINGSSVGLKSGSLK
TQEDTPSAPPPRTFINQLPDWSMLLAAITTVFLAAEKQWMMLDWKPKRPDMLVDPFGLGRIVQDGLVFRQ
NFSIRSYEIGADRTASIETVMNHLQETALNHVKSAGLLNDGFGRTLEMYKRDLIWVVAKMQVMVNRYPTW
GDTVEVNTWVAKSGKNGMRRDWLISDCNTGEILTRASSKSQIMLPLHYCSVWVMMNQKTRRLSKIPDEVR
HEIEPHFVDSAPVIEDDDRKLPKLDEKTADSIRKGLTPKWNDLDVNQHVNNVKYIGWILESTPPEVLETQ
ELCSLTLEYRRECGRESVLESLTAVDPSGKGSGSQFQHLLRLEDGGEIVKGRTEWRPKTAGINGPIASGE
TSPGDSS*
SEQ ID NO: 17
Cuphea hyssopifolia (Chs) FATB2b +a.a.248-259 variant coding DNA
sequence
ATGGTGGCTACCGCTGCAAGTTCAGCATTCTTCCCTGTGCCGTCCCCCGACGCCTCCTCTAGACCTGGAA
AGCTCGGCAATGGGTCATCGAGCTTGAGCCCCCTCAAGCCCAAATTGATGGCCAATGGCGGGTTGCAGGT
TAAGGCAAACGCCAGTGCCCCTCCTAAGATCAATGGTTCTTCGGTCGGTCTAAAGTCCGGCAGTCTCAAG
ACTCAGGAAGACACTCCTTCGGCGCCTCCTCCdCGGACTTTTATTAACCAGCTGCCTGATTGGAGTATGC
TTCTTGCTGCAATCACTACTGTCTTCTTGGCAGCAGAGAAGCAGTGGATGATGCTTGATTGGAAACCCAA
GAGGCCTGACATGCTTGTGGACCCGTTCGGATTGGGAAGGATTGTTCAAGATGGGCTTGTGTTCAGGCAG
AATTTTTCGATTAGGTCCTATGAAATAGGCGCTGATCGCACTGCGTCTATAGAGACGGTGATGAACCACT
TGCAGGAAACAGCTCTCAATCATGTTAAGAGTGCTGGGCTTCTTAATGACGGCTTTGGTCGTACTCTTGA
GATGTATAAAAGGGACCTTATTTGGGTTGTTGCAAAAATGCAGGTCATGGTTAACCGCTATCCTACTTGG
GGCGACACGGTTGAAGTGAATACTTGGGTTGCCAAGTCAGGGAAAAATGGTATGCGTCGTGATTGGCTCA
TAAGTGATTGCAATACAGGAGAAATTCTTACTAGAGCATCAAGTAAAAGCCAAATTATGTTACCCTTACA
TTATTGCAGTGTGTGGGTCATGATGAATCAAAAGACAAGAAGATTGTCAAAAATTCCAGATGAGGTTCGA
CATGAGATAGAGCCTCATTTCGTGGACTCTGCTCCCGTCATTGAAGATGATGACCGGAAACTTCCCAAGC
TGGATGAGAAGACTGCTGACTCCATCCGCAAGGGTCTAACTCCGAAGTGGAATGACTTGGATGTCAATCA
GCACGTCAACAACGTGAAGTACATTGGGTGGATTCTTGAGAGTACTCCACCAGAAGTTCTGGAGACCCAG
GAGTTATGTTCCCTTACCCTGGAATATAGGCGGGAATGCGGAAGGGAGAGCGTGCTGGAGTCCCTCACTG
CTGTGGACCCCTCTGGAAAGGGCTCTGGGTCTCAGTTCCAGCACCTTCTGCGGCTTGAGGATGGAGGTGA
GATTGTGAAGGGGAGAACTGAGTGGCGACCCAAGACTGCAGGAATCAATGGGCCAATAGCATCCGGGGAG
ACCTCACCTGGAGACTCTTCTTAG
SEQ ID NO: 18
Cuphea hyssopifolia (Chs) FATB2b +a.a.248-259 variant coding DNA
sequence codon optimized for Prototheca moriformis
ATGGTGGCCACCGCCGCCTCCTCCGCCTTCTTCCCCGTGCCCTCCCCCGACGCCTCCTCCCGCCCCGGCA
AGCTGGGCAACGGCTCCTCCTCCCTGTCCCCCCTGAAGCCCAAGCTGATGGCCAACGGCGGCCTGCAGGT
GAAGGCCAACGCCTCCGCCCCCCCCAAGATCAACGGCTCCTCCGTGGGCCTGAAGTCCGGCTCCCTGAAG
ACCCAGGAGGACACCCCCTCCGCCCCCCCCCCCCGCACCTTCATCAACCAGCTGCCCGACTGGTCCATGC
TGCTGGCCGCCATCACCACCGTGTTCCTGGCCGCCGAGAAGCAGTGGATGATGCTGGACTGGAAGCCCAA
GCGCCCCGACATGCTGGTGGACCCCTTCGGCCTGGGCCGCATCGTGCAGGACGGCCTGGTGTTCCGCCAG
AACTTCTCCATCCGCTCCTACGAGATCGGCGCCGACCGCACCGCCTCCATCGAGACCGTGATGAACCACC
TGCAGGAGACCGCCCTGAACCACGTGAAGTCCGCCGGCCTGCTGAACGACGGCTTCGGCCGCACCCTGGA
GATGTACAAGCGCGACCTGATCTGGGTGGTGGCCAAGATGCAGGTGATGGTGAACCGCTACCCCACCTGG
GGCGACACCGTGGAGGTGAACACCTGGGTGGCCAAGTCCGGCAAGAACGGCATGCGCCGCGACTGGCTGA
TCTCCGACTGCAACACCGGCGAGATCCTGACCCGCGCCTCCTCCAAGTCCCAGATCATGCTGCCCCTGCA
CTACTGCTCCGTGTGGGTGATGATGAACCAGAAGACCCGCCGCCTGTCCAAGATCCCCGACGAGGTGCGC
CACGAGATCGAGCCCCACTTCGTGGACTCCGCCCCCGTGATCGAGGACGACGACCGCAAGCTGCCCAAGC
TGGACGAGAAGACCGCCGACTCCATCCGCAAGGGCCTGACCCCCAAGTGGAACGACCTGGACGTGAACCA
GCACGTGAACAACGTGAAGTACATCGGCTGGATCCTGGAGTCCACCCCCCCCGAGGTGCTGGAGACCCAG
GAGCTGTGCTCCCTGACCCTGGAGTACCGCCGCGAGTGCGGCCGCGAGTCCGTGCTGGAGTCCCTGACCG
CCGTGGACCCCTCCGGCAAGGGCTCCGGCTCCCAGTTCCAGCACCTGCTGCGCCTGGAGGACGGCGGCGA
GATCGTGAAGGGCCGCACCGAGTGGCGCCCCAAGACCGCCGGCATCAACGGCCCCATCGCCTCCGGCGAG
ACCTCCCCCGGCGACTCCTCCTGA
SEQ ID NO: 19
Cuphea hyssopifolia (Chs) FATB3 amino acid sequence
MVAAEASSALFSVRTPGTSPKPGKFGNWPTSLSVPFKSKSNHNGGFQVKANASARPKANGSAVSLKSGSL
DTQEDTSSSSSPPRTFINQLPDWSMLLSAITTVFVAAEKQWTMLDRKSKRPDMLMDPFGVDRVVQDGAVF
RQSFSIRSYEIGADRTASIETLMNIFQETSLNHCKSIGLLNDGFGRTPEMCKRDLIWVVTKMHVEVNRYP
TWGDTIEVNTWVSESGKTGMGRDWLISDCHTGEILIRATSMCAMMNQKTRRFSKFPYEVRQELAPHFVDS
APVIEDYQKLHKLDVKTGDSICNGLTPRWNDLDVNQHVNNVKYIGWILESVPTEVFETQELCGLTLEYRR
ECGRDSVLESVTAMDPSKEGDRSLYQHLLRLEDGADIAKGRTKWRPKNAGTNGAISTGKTSNGNSIS*
SEQ ID NO: 20
Cuphea hyssopifolia (Chs) FATB3 coding DNA sequence
ATGGTGGCTGCCGAAGCAAGTTCTGCACTCTTCTCCGTTCGAACCCCGGGAACCTCCCCTAAACCCGGGA
AGTTCGGGAATTGGCCAACGAGCTTGAGCGTCCCCTTCAAGTCCAAATCAAACCACAATGGCGGCTTTCA
GGTTAAGGCAAACGCCAGTGCCCGTCCTAAGGCTAACGGTTCTGCAGTAAGTCTAAAGTCTGGCAGCCTC
GACACTCAGGAGGACACTTCATCGTCGTCCTCTCCTCCTCGGACTTTCATTAACCAGTTGCCCGACTGGA
GTATGCTGCTGTCCGCGATCACGACCGTCTTCGTGGCGGCTGAGAAGCAGTGGACGATGCTTGATCGGAA
ATCTAAGAGGCCCGACATGCTCATGGACCCGTTTGGGGTTGACAGGGTTGTTCAGGATGGGGCTGTGTTC
AGACAGAGTTTTTCGATTAGGTCTTACGAAATAGGCGCTGATCGAACAGCCTCTATAGAGACGCTGATGA
ACATCTTCCAGGAAACATCTCTCAATCATTGTAAGAGTATCGGTCTTCTCAATGACGGCTTTGGTCGTAC
TCCTGAGATGTGTAAGAGGGACCTCATTTGGGTGGTTACAAAAATGCACGTCGAGGTTAATCGCTATCCT
ACTTGGGGTGATACTATCGAGGTCAATACTTGGGTCTCCGAGTCGGGGAAAACCGGTATGGGTCGTGATT
GGCTGATAAGTGATTGTCATACAGGAGAAATTCTAATAAGAGCAACGAGCATGTGTGCTATGATGAATCA
AAAGACGAGAAGATTCTCAAAATTTCCATATGAGGTTCGACAGGAGTTGGCGCCTCATTTTGTGGACTCT
GCTCCTGTCATTGAAGACTATCAAAAATTGCACAAGCTTGATGTGAAGACGGGTGATTCCATTTGCAATG
GCCTAACTCCAAGGTGGAATGACTTGGATGTCAATCAGCACGTTAACAATGTGAAGTACATTGGGTGGAT
TCTCGAGAGTGTTCCAACGGAAGTTTTCGAGACCCAGGAGCTATGTGGCCTCACCCTTGAGTATAGGCGG
GAATGCGGAAGGGACAGTGTGCTGGAGTCCGTGACCGCTATGGATCCATCAAAAGAGGGAGACAGATCTC
TGTACCAGCACCTTCTTCGGCTTGAGGATGGGGCTGATATCGCGAAGGGCAGAACCAAGTGGCGGCCGAA
GAATGCAGGAACCAATGGGGCAATATCAACAGGAAAGACTTCAAATGGAAACTCGATCTCTTAG
SEQ ID NO: 21
Cuphea hyssopifolia (Chs) FATB3 coding DNA sequence codon optimized
for Prototheca moriformis
ATGGTGGCCGCCGAGGCCTCCTCCGCCCTGTTCTCCGTGCGCACCCCCGGCACCTCCCCCAAGCCCGGCA
AGTTCGGCAACTGGCCCACCTCCCTGTCCGTGCCCTTCAAGTCCAAGTCCAACCACAACGGCGGCTTCCA
GGTGAAGGCCAACGCCTCCGCCCGCCCCAAGGCCAACGGCTCCGCCGTGTCCCTGAAGTCCGGCTCCCTG
GACACCCAGGAGGACACCTCCTCCTCCTCCTCCCCCCCCCGCACCTTCATCAACCAGCTGCCCGACTGGT
CCATGCTGCTGTCCGCCATCACCACCGTGTTCGTGGCCGCCGAGAAGCAGTGGACCATGCTGGACCGCAA
GTCCAAGCGCCCCGACATGCTGATGGACCCCTTCGGCGTGGACCGCGTGGTGCAGGACGGCGCCGTGTTC
CGCCAGTCCTTCTCCATCCGCTCCTACGAGATCGGCGCCGACCGCACCGCCTCCATCGAGACCCTGATGA
ACATCTTCCAGGAGACCTCCCTGAACCACTGCAAGTCCATCGGCCTGCTGAACGACGGCTTCGGCCGCAC
CCCCGAGATGTGCAAGCGCGACCTGATCTGGGTGGTGACCAAGATGCACGTGGAGGTGAACCGCTACCCC
ACCTGGGGCGACACCATCGAGGTGAACACCTGGGTGTCCGAGTCCGGCAAGACCGGCATGGGCCGCGACT
GGCTGATCTCCGACTGCCACACCGGCGAGATCCTGATCCGCGCCACCTCCATGTGCGCCATGATGAACCA
GAAGACCCGCCGCTTCTCCAAGTTCCCCTACGAGGTGCGCCAGGAGCTGGCCCCCCACTTCGTGGACTCC
GCCCCCGTGATCGAGGACTACCAGAAGCTGCACAAGCTGGACGTGAAGACCGGCGACTCCATCTGCAACG
GCCTGACCCCCCGCTGGAACGACCTGGACGTGAACCAGCACGTGAACAACGTGAAGTACATCGGCTGGAT
CCTGGAGTCCGTGCCCACCGAGGTGTTCGAGACCCAGGAGCTGTGCGGCCTGACCCTGGAGTACCGCCGC
GAGTGCGGCCGCGACTCCGTGCTGGAGTCCGTGACCGCCATGGACCCCTCCAAGGAGGGCGACCGCTCCC
TGTACCAGCACCTGCTGCGCCTGGAGGACGGCGCCGACATCGCCAAGGGCCGCACCAAGTGGCGCCCCAA
GAACGCCGGCACCAACGGCGCCATCTCCACCGGCAAGACCTCCAACGGCAACTCCATCTCCTGA
SEQ ID NO: 22
Cuphea hyssopifolia (Chs) FATB3b (V204I, C239F, E243D, M251V variant)
amino acid sequence
MVAAEASSALFSVRTPGTSPKPGKFGNWPTSLSVPFKSKSNHNGGFQVKANASARPKANGSAVSLKSGSL
DTQEDTSSSSSPPRTFINQLPDWSMLLSAITTVFVAAEKQWTMLDRKSKRPDMLMDPFGVDRVVQDGAVF
RQSFSIRSYEIGADRTASIETLMNIFQETSLNHCKSIGLLNDGFGRTPEMCKRDLIWVVTKMHIEVNRYP
TWGDTIEVNTWVSESGKTGMGRDWLISDFHTGDILIRATSVCAMMNQKTRRFSKFPYEVRQELAPHFVDS
APVIEDYQKLHKLDVKTGDSICNGLTPRWNDLDVNQHVNNVKYIGWILESVPTEVFETQELCGLTLEYRR
ECGRDSVLESVTAMDPSKEGDRSLYQHLLRLEDGADIAKGRTKWRPKNAGTNGAISTGKTSNGNSIS*
SEQ ID NO: 23
Cuphea hyssopifolia (Chs) FATB3b (V204I, C239F, E243D, M251V variant)
coding DNA sequence
ATGGTGGCTGCCGAAGCAAGTTCTGCACTCTTCTCCGTTCGAACCCCGGGAACCTCCCCTAAACCCGGGA
AGTTCGGGAATTGGCCAACGAGCTTGAGCGTCCCCTTCAAGTCCAAATCAAACCACAATGGCGGCTTTCA
GGTTAAGGCAAACGCCAGTGCCCGTCCTAAGGCTAACGGTTCTGCAGTAAGTCTAAAGTCTGGCAGCCTC
GACACTCAGGAGGACACTTCATCGTCGTCCTCTCCTCCTCGGACTTTCATTAACCAGTTGCCCGACTGGA
GTATGCTGCTGTCCGCGATCACGACCGTCTTCGTGGCGGCTGAGAAGCAGTGGACGATGCTTGATCGGAA
ATCTAAGAGGCCCGACATGCTCATGGACCCGTTTGGGGTTGACAGGGTTGTTCAGGATGGGGCTGTGTTC
AGACAGAGTTTTTCGATTAGGTCTTACGAAATAGGCGCTGATCGAACAGCCTCTATAGAGACGCTGATGA
ACATCTTCCAGGAAACATCTCTCAATCATTGTAAGAGTATCGGTCTTCTCAATGACGGCTTTGGTCGTAC
TCCTGAGATGTGTAAGAGGGACCTCATTTGGGTGGTTACAAAAATGCACATCGAGGTTAATCGCTATCCT
ACTTGGGGTGATACTATCGAGGTCAATACTTGGGTCTCCGAGTCGGGGAAAACCGGTATGGGTCGTGATT
GGCTGATAAGTGATTTTCATACAGGAGACATTCTAATAAGAGCAACGAGCGTGTGTGCTATGATGAATCA
AAAGACGAGAAGATTCTCAAAATTTCCATATGAGGTTCGACAGGAGTTAGCGCCTCATTTTGTGGACTCT
GCTCCAGTCATTGAAGACTATCAAAAATTGCACAAGCTTGATGTGAAGACGGGTGATTCCATTTGCAATG
GCCTAACTCCAAGGTGGAATGACTTGGATGTCAATCAGCACGTTAACAATGTGAAGTACATTGGGTGGAT
TCTCGAGAGTGTTCCAACGGAAGTTTTCGAGACCCAGGAGCTATGTGGCCTCACCCTTGAGTATAGGCGG
GAATGCGGAAGGGACAGTGTGCTGGAGTCCGTGACCGCTATGGATCCCTCAAAAGAGGGAGACAGATCTC
TGTACCAGCACCTTCTTCGGCTTGAGGATGGGGCTGATATCGCGAAGGGCAGAACCAAGTGGCGGCCGAA
GAATGCAGGAACCAATGGGGCAATATCAACAGGAAAGACTTCAAATGGAAACTCGATCTCTTAG
SEQ ID NO: 24
Cuphea hyssopifolia (Chs) FATB3b (V204I, C239F, E243D, M251V variant)
coding DNA sequence codon optimized for Prototheca moriformis
ATGGTGGCCGCCGAGGCCTCCTCCGCCCTGTTCTCCGTGCGCACCCCCGGCACCTCCCCCAAGCCCGGCA
AGTTCGGCAACTGGCCCACCTCCCTGTCCGTGCCCTTCAAGTCCAAGTCCAACCACAACGGCGGCTTCCA
GGTGAAGGCCAACGCCTCCGCCCGCCCCAAGGCCAACGGCTCCGCCGTGTCCCTGAAGTCCGGCTCCCTG
GACACCCAGGAGGACACCTCCTCCTCCTCCTCCCCCCCCCGCACCTTCATCAACCAGCTGCCCGACTGGT
CCATGCTGCTGTCCGCCATCACCACCGTGTTCGTGGCCGCCGAGAAGCAGTGGACCATGCTGGACCGCAA
GTCCAAGCGCCCCGACATGCTGATGGACCCCTTCGGCGTGGACCGCGTGGTGCAGGACGGCGCCGTGTTC
CGCCAGTCCTTCTCCATCCGCTCCTACGAGATCGGCGCCGACCGCACCGCCTCCATCGAGACCCTGATGA
ACATCTTCCAGGAGACCTCCCTGAACCACTGCAAGTCCATCGGCCTGCTGAACGACGGCTTCGGCCGCAC
CCCCGAGATGTGCAAGCGCGACCTGATCTGGGTGGTGACCAAGATGCACATCGAGGTGAACCGCTACCCC
ACCTGGGGCGACACCATCGAGGTGAACACCTGGGTGTCCGAGTCCGGCAAGACCGGCATGGGCCGCGACT
GGCTGATCTCCGACTTCCACACCGGCGACATCCTGATCCGCGCCACCTCCGTGTGCGCCATGATGAACCA
GAAGACCCGCCGCTTCTCCAAGTTCCCCTACGAGGTGCGCCAGGAGCTGGCCCCCCACTTCGTGGACTCC
GCCCCCGTGATCGAGGACTACCAGAAGCTGCACAAGCTGGACGTGAAGACCGGCGACTCCATCTGCAACG
GCCTGACCCCCCGCTGGAACGACCTGGACGTGAACCAGCACGTGAACAACGTGAAGTACATCGGCTGGAT
CCTGGAGTCCGTGCCCACCGAGGTGTTCGAGACCCAGGAGCTGTGCGGCCTGACCCTGGAGTACCGCCGC
GAGTGCGGCCGCGACTCCGTGCTGGAGTCCGTGACCGCCATGGACCCCTCCAAGGAGGGCGACCGCTCCC
TGTACCAGCACCTGCTGCGCCTGGAGGACGGCGCCGACATCGCCAAGGGCCGCACCAAGTGGCGCCCCAA
GAACGCCGGCACCAACGGCGCCATCTCCACCGGCAAGACCTCCAACGGCAACTCCATCTCCTGA
SEQ ID NO: 25
Cuphea PSR23 (Cu) FATB3 amino acid sequence
MVVAAATSAFFPVPAPGTSPKPGKSGNWPSSLSPTFKPKSIPNAGFQVKANASAHPKANGSAVNLKSGSL
NTQEDTSSSPPPRAFLNQLPDWSMLLTAITTVFVAAEKQWTMLDRKSKRPDMLVDSVGLKCIVRDGLVSR
QSFLIRSYEIGADRTASIETLMNHLQETSINHCKSLGLLNDGFGRTPGMCKNDLIWVLTKMQIMVNRYPT
WGDTVEINTWFSQSGKIGMASDWLISDCNTGEILIRATSVWAMMNQKTRRFSRLPYEVRQELTPHFVDSP
HVIEDNDQKLHKFDVKTGDSIRKGLTPRWNDLDVNQHVSNVKYIGWILESMPIEVLETQELCSLTVEYRR
ECGMDSVLESVTAVDPSENGGRSQYKHLLRLEDGTDIVKSRTEWRPKNAGTNGAISTSTAKTSNGNSVS*
SEQ ID NO: 26
Cuphea PSR23 (Cu) FATB3 coding DNA sequence
ATGGTGGTGGCTGCAGCAACTTCTGCATTCTTCCCCGTTCCAGCCCCGGGAACCTCCCCTAAACCCGGGA
AGTCCGGCAACTGGCCATCGAGCTTGAGCCCTACCTTCAAGCCCAAGTCAATCCCCAATGCCGGATTTCA
GGTTAAGGCAAATGCCAGTGCCCATCCTAAGGCTAACGGTTCTGCAGTAAATCTAAAGTCTGGCAGCCTC
AACACTCAGGAGGACACTTCGTCGTCCCCTCCTCCCCGGGCTTTCCTTAACCAGTTGCCTGATTGGAGTA
TGCTTCTGACTGCAATCACGACCGTCTTCGTGGCGGCAGAGAAGCAGTGGACTATGCTTGATAGGAAATC
TAAGAGGCCTGACATGCTCGTGGACTCGGTTGGGTTGAAGTGTATTGTTCGGGATGGGCTCGTGTCCAGA
CAGAGTTTTTTGATTAGATCTTATGAAATAGGCGCTGATCGAACAGCCTCTATAGAGACGCTGATGAACC
ACTTGCAGGAAACATCTATCAATCATTGTAAGAGTTTGGGTCTTCTCAATGACGGCTTTGGTCGTACTCC
TGGGATGTGTAAAAACGACCTCATTTGGGTGCTTACAAAAATGCAGATCATGGTGAATCGCTACCCAACT
TGGGGCGATACTGTTGAGATCAATACCTGGTTCTCTCAGTCGGGGAAAATCGGTATGGCTAGCGATTGGC
TAATAAGTGATTGCAACACAGGAGAAATTCTTATAAGAGCAACGAGCGTGTGGGCTATGATGAATCAAAA
GACGAGAAGATTCTCAAGACTTCCATACGAGGTTCGCCAGGAGTTAACGCCTCATTTTGTGGACTCTCCT
CATGTCATTGAAGACAATGATCAGAAATTGCATAAGTTTGATGTGAAGACTGGTGATTCCATTCGCAAGG
GTCTAACTCCGAGGTGGAACGACTTGGATGTGAATCAGCACGTAAGCAACGTGAAGTACATTGGGTGGAT
TCTCGAGAGTATGCCAATAGAAGTTTTGGAGACACAGGAGCTATGCTCTCTCACCGTAGAATATAGGCGG
GAATGCGGAATGGACAGTGTGCTGGAGTCCGTGACTGCTGTGGATCCCTCAGAAAATGGAGGCCGGTCTC
AGTACAAGCACCTTCTGCGGCTTGAGGATGGGACTGATATCGTGAAGAGCAGAACTGAGTGGCGACCGAA
GAATGCAGGAACTAACGGGGCGATATCAACATCAACAGCAAAGACTTCAAATGGAAACTCGGTCTCTTAG
SEQ ID NO: 27
Cuphea PSR23 (Cu) FATB3 coding DNA sequence codon optimized for
Prototheca moriformis
ATGGTGGTGGCCGCCGCCACCTCCGCCTTCTTCCCCGTGCCCGCCCCCGGCACCTCCbCCAAGCCCGGCA
AGTCCGGCAACTGGCCCTCCTCCCTGTCCCCCACCTTCAAGCCCAAGTCCATCCCCAACGCCGGCTTCCA
GGTGAAGGCCAACGCCTCCGCCCACCCCAAGGCCAACGGCTCCGCCGTGAACCTGAAGTCCGGCTCCCTG
AACACCCAGGAGGACACCTCCTCCTCCCCCCCCCCCCGCGCCTTCCTGAACCAGCTGCCCGACTGGTCCA
TGCTGCTGACCGCCATCACCACCGTGTTCGTGGCCGCCGAGAAGCAGTGGACCATGCTGGACCGCAAGTC
CAAGCGCCCCGACATGCTGGTGGACTCCGTGGGCCTGAAGTGCATCGTGCGCGACGGCCTGGTGTCCCGC
CAGTCCTTCCTGATCCGCTCCTACGAGATCGGCGCCGACCGCACCGCCTCCATCGAGACCCTGATGAACC
ACCTGCAGGAGACCTCCATCAACCACTGCAAGTCCCTGGGCCTGCTGAACGACGGCTTCGGCCGCACCCC
CGGCATGTGCAAGAACGACCTGATCTGGGTGCTGACCAAGATGCAGATCATGGTGAACCGCTACCCCACC
TGGGGCGACACCGTGGAGATCAACACCTGGTTCTCCCAGTCCGGCAAGATCGGCATGGCCTCCGACTGGC
TGATCTCCGACTGCAACACCGGCGAGATCCTGATCCGCGCCACCTCCGTGTGGGCCATGATGAACCAGAA
GACCCGCCGCTTCTCCCGCCTGCCCTACGAGGTGCGCCAGGAGCTGACCCCCCACTTCGTGGACTCCCCC
CACGTGATCGAGGACAACGACCAGAAGCTGCACAAGTTCGACGTGAAGACCGGCGACTCCATCCGCAAGG
GCCTGACCCCCCGCTGGAACGACCTGGACGTGAACCAGCACGTGTCCAACGTGAAGTACATCGGCTGGAT
CCTGGAGTCCATGCCCATCGAGGTGCTGGAGACCCAGGAGCTGTGCTCCCTGACCGTGGAGTACCGCCGC
GAGTGCGGCATGGACTCCGTGCTGGAGTCCGTGACCGCCGTGGACCCCTCCGAGAACGGCGGCCGCTCCC
AGTACAAGCACCTGCTGCGCCTGGAGGACGGCACCGACATCGTGAAGTCCCGCACCGAGTGGCGCCCCAA
GAACGCCGGCACCAACGGCGCCATCTCCACCTCCACCGCCAAGACCTCCAACGGCAACTCCGTGTCCTGA
SEQ ID NO: 28
Cuphea wrightii (Cw) FATB3 amino acid sequence
MVVAAAASSAFFPVPAPRTTPKPGKFGNWPSSLSPPFKPKSNPNGRFQVKANVSPHPKANGSAVSLKSGS
LNTLEDPPSSPPPRTFLNQLPDWSRLRTAITTVFVAAEKQFTRLDRKSKRPDMLVDWFGSETIVQDGLVF
RERFSIRSYEIGADRTASIETLMNHLQDTSLNHCKSVGLLNDGFGRTSEMCTRDLIWVLTKMQIVVNRYP
TWGDTVEINSWFSQSGKIGMGRDWLISDCNTGEILVRATSAWAMMNQKTRRFSKLPCEVRQEIAPHFVDA
PPVIEDNDRKLHKFDVKTGDSICKGLTPGWNDLDVNQHVSNVKYIGWILESMPTEVLETQELCSLTLEYR
RECGRESVVESVTSMNPSKVGDRSQYQHLLRLEDGADIMKGRTEWRPKNAGTNRAIST*
SEQ ID NO: 29
Cuphea wrightii (Cw) FATB3 coding DNA sequence
ATGGTGGTGGCTGCTGCAGCAAGTTCTGCATTCTTCCCTGTTCCAGCACCTAGAACCACGCCTAAACCCG
GGAAGTTCGGCAATTGGCCATCGAGCTTGAGCCCGCCCTTCAAGCCCAAGTCAAACCCCAATGGTAGATT
TCAGGTTAAGGCAAATGTCAGTCCTCATCCTAAGGCTAACGGTTCTGCAGTAAGTCTAAAGTCTGGCAGC
CTCAACACTCTGGAGGACCCTCCGTCGTCCCCTCCTCCTCGGACTTTCCTTAACCAGTTGCCTGATTGGA
GTAGGCTTCGGACTGCAATCACGACCGTCTTCGTGGCGGCAGAGAAGCAGTTCACTAGGCTCGATCGAAA
ATCTAAGAGGCCTGACATGCTAGTGGACTGGTTTGGGTCAGAGACTATTGTTCAGGATGGGCTCGTGTTC
AGAGAGAGATTTTtGATCAGGTCTTACGAAATAGGCGCTGATCGAACAGCCTCTATAGAGACGCTGATGA
ACCACTTGCAGGACACATCTCTGAATCATTGTAAGAGTGTGGGTCTTCTCAATGACGGCTTTGGTCGTAC
CTCGGAGATGTGTACAAGAGACCTCATTTGGGTGCTTACAAAAATGCAGATCGTGGTGAATCGCTATCCA
ACTTGGGGCGATACTGTCGAGATCAATAGCTGGTTCTCCCAGTCGGGGAAAATCGGTATGGGTCGCGATT
GGCTAATAAGTGATTGCAACACAGGAGAAATTCTTGTAAGAGCAACGAGCGCTTGGGCCATGATGAATCA
AAAGACGAGAAGATTCTCAAAACTTCCATGCGAGGTTCGCCAGGAGATAGCGCCTCATTTTGTGGACGCT
CCTCCTGTCATTGAAGACAATGATCGGAAATTGCATAAGTTTGATGTGAAGACTGGTGATTCCATTTGCA
AGGGTCTAACTCCGGGGTGGAATGACTTGGATGTCAATCAGCACGTAAGCAACGTGAAGTACATTGGGTG
GATTCTCGAGAGTATGCCTACAGAAGTTTTGGAGACCCAGGAGCTATGCTCTCTCACCCTTGAATATAGG
CGGGAATGTGGAAGGGAAAGTGTGGTAGAGTCCGTGACCTCTATGAATCCCTCAAAAGTTGGAGACCGGT
CTCAGTACCAACACCTTCTGCGGCTTGAGGATGGGGCTGATATCATGAAGGGCAGAACTGAGTGGAGACC
AAAGAATGCAGGAACCAACCGGGCGATATCAACATGA
SEQ ID NO: 30
Cuphea wrightii (Cw) FATB3 coding DNA sequence codon optimized for
Prototheca moriformis
ATGGTGGTGGCCGCCGCCGCCTCCTCCGCCTTCTTCCCCGTGCCCGCCCCCCGCACCACCCCCAAGCCCG
GCAAGTTCGGCAACTGGCCCTCCTCCCTGTCCCCCCCCTTCAAGCCCAAGTCCAACCCCAACGGCCGCTT
CCAGGTGAAGGCCAACGTGTCCCCCCACCCCAAGGCCAACGGCTCCGCCGTGTCCCTGAAGTCCGGCTCC
CTGAACACCCTGGAGGACCCCCCCTCCTCCCCCCCCCCCCGCACCTTCCTGAACCAGCTGCCCGACTGGT
CCCGCCTGCGCACCGCCATCACCACCGTGTTCGTGGCCGCCGAGAAGCAGTTCACCCGCCTGGACCGCAA
GTCCAAGCGCCCCGACATGCTGGTGGACTGGTTCGGCTCCGAGACCATCGTGCAGGACGGCCTGGTGTTC
CGCGAGCGCTTCTCCATCCGCTCCTACGAGATCGGCGCCGACCGCACCGCCTCCATCGAGACCCTGATGA
ACCACCTGCAGGACACCTCCCTGAACCACTGCAAGTCCGTGGGCCTGCTGAACGACGGCTTCGGCCGCAC
CTCCGAGATGTGCACCCGCGACCTGATCTGGGTGCTGACCAAGATGCAGATCGTGGTGAACCGCTACCCC
ACCTGGGGCGACACCGTGGAGATCAACTCCTGGTTCTCCCAGTCCGGCAAGATCGGCATGGGCCGCGACT
GGCTGATCTCCGACTGCAACACCGGCGAGATCCTGGTGCGCGCCACCTCCGCCTGGGCCATGATGAACCA
GAAGACCCGCCGCTTCTCCAAGCTGCCCTGCGAGGTGCGCCAGGAGATCGCCCCCCACTTCGTGGACGCC
CCCCCCGTGATCGAGGACAACGACCGCAAGCTGCACAAGTTCGACGTGAAGACCGGCGACTCCATCTGCA
AGGGCCTGACCCCCGGCTGGAACGACCTGGACGTGAACCAGCACGTGTCCAACGTGAAGTACATCGGCTG
GATCCTGGAGTCCATGCCCACCGAGGTGCTGGAGACCCAGGAGCTGTGCTCCCTGACCCTGGAGTACCGC
CGCGAGTGCGGCCGCGAGTCCGTGGTGGAGTCCGTGACCTCCATGAACCCCTCCAAGGTGGGCGACCGCT
CCCAGTACCAGCACCTGCTGCGCCTGGAGGACGGCGCCGACATCATGAAGGGCCGCACCGAGTGGCGCCC
CAAGAACGCCGGCACCAACCGCGCCATCTCCACCTGA
SEQ ID NO: 31
Cuphea wrightii (Cw) FATB4a amino acid sequence
MVATAASSAFFPVPSADTSSSRPGKLGSGPSSLSPLKPKSIPNGGLQVKANASAPPKINGSSVGLKSGGF
KTQEDSPSAPPPRTFINQLPDWSMLLAAITTVFLAAEKQWMMLDWKPKRPDMLVDPFGLGSIVQDGLVFR
QNFSIRSYEIGADRTASIETVMNHLQETALNHVKIAGLSNDGFGRTPEMYKRDLIWVVAKMQVMVNRYPT
WGDTVEVNTWVAKSGKNGMRRDWLISDCNTGEILTRASSVWVMMNQKTRRLSKIPDEVRNEIEPHFVDSA
PVVEDDDRKLPKLDENTADSIRKGLTPRWNDLDVNQHVNNVKYIGWILESTPPEVLETQELCSLTLEYRR
ECGRESVLESLTAVDPSAEGYASRFQHLLRLEDGGEIVKARTEWRPKNAGINGVVPSEESSPGDFF*
SEQ ID NO: 32
Cuphea wrightii (Cw) FATB4a coding DNA sequence
TTGGTGGCTACCGCTGCAAGTTCTGCATTTTTCCCCGTGCCATCCGCCGACACCTCCTCCTCGAGACCCG
GAAAGCTCGGCAGTGGACCATCGAGCTTGAGCCCCCTCAAGCCCAAATCGATCCCCAATGGCGGCTTGCA
GGTTAAGGCAAACGCCAGTG6CCCTCCTAAGATCAATGGTTCCTCGGTCGGTCTAAAGTCGGGCGGTTTC
AAGACTCAGGAAGACTCTCCTTCGGCCCCTCCTCCGCGGACTTTTATCAACCAGTTGCCTGATTGGAGTA
TGCTTCTTGCTGCAATCACTACTGTCTTCTTGGCTGCAGAGAAGCAGTGGATGATGCTTGATTGGAAACC
TAAGAGGCCTGACATGCTCGTGGACCCGTTCGGATTGGGAAGTATTGTTCAGGATGGGCTTGTGTTCAGG
CAGAATTTTTCAATTAGGTCCTACGAAATAGGCGCCGATCGAACTGCGTCTATAGAGACGGTGATGAACC
ATTTGCAGGAAACAGCTCTCAATCATGTCAAGATTGCTGGGCTTTCTAATGACGGCTTTGGTCGTACTCC
TGAGATGTATAAAAGAGACCTTATTTGGGTTGTTGCAAAAATGCAGGTCATGGTTAACCGCTATCCTACT
TGGGGTGACACGGTTGAAGTGAATACTTGGGTTGCCAAGTCAGGGAAAAATGGTATGCGTCGTGACTGGC
TCATAAGTGATTGCAATACTGGAGAGATTCTTACAAGAGCATCAAGCGTGTGGGTCATGATGAATCAAAA
GACAAGAAGATTGTCAAAAATTCCAGATGAGGTTCGAAATGAGATAGAGCCTCATTTTGTGGACTCTGCT
CCCGTCGTTGAAGATGATGATCGGAAACTTCCCAAGCTGGATGAGAACACTGCTGACTCCATCCGCAAGG
GTCTAACTCCGAGGTGGAATGACTTGGATGTCAATCAGCACGTCAACAACGTGAAGTACATCGGATGGAT
TCTTGAGAGTACTCCACCAGAAGTTCTGGAGACCCAGGAGTTATGCTCCCTGACCCTGGAATACAGGCGG
GAATGTGGAAGGGAGAGCGTGCTGGAGTCCCTCACTGCTGTCGACCCGTCTGCAGAGGGCTATGCGTCCC
GGTTTCAGCACCTTCTGCGGCTTGAGGATGGAGGTGAGATCGTGAAGGCGAGAACTGAGTGGCGACCCAA
GAATGCTGGAATCAATGGGGTGGTACCATCCGAGGAGTCCTCACCTGGAGACTTCTTTTAG
SEQ ID NO: 33
Cuphea wrightii (Cw) FATB4a coding DNA sequence codon optimized for
Prototheca moriformis
ATGGTGGCCACCGCCGCCTCCTCCGCCTTCTTCCCCGTGCCCTCCGCCGACACCTCCTCCTCCCGCCCCG
GCAAGCTGGGCTCCGGCCCCTCCTCCCTGTCCCCCCTGAAGCCCAAGTCCATCCCCAACGGCGGCCTGCA
GGTGAAGGCCAACGCCTCCGCCCCCCCCAAGATCAACGGCTCCTCCGTGGGCCTGAAGTCCGGCGGCTTC
AAGACCCAGGAGGACTCCCCCTCCGCCCCCCCCCCCCGCACCTTCATCAACCAGCTGCCCGACTGGTCCA
TGCTGCTGGCCGCCATCACCACCGTGTTCCTGGCCGCCGAGAAGCAGTGGATGATGCTGGACTGGAAGCC
CAAGCGCCCCGACATGCTGGTGGACCCCTTCGGCCTGGGCTCCATCGTGCAGGACGGCCTGGTGTTCCGC
CAGAACTTCTCCATCCGCTCCTACGAGATCGGCGCCGACCGCACCGCCTCCATCGAGACCGTGATGAACC
ACCTGCAGGAGACCGCCCTGAACCACGTGAAGATCGCCGGCCTGTCCAACGACGGCTTCGGCCGCACCCC
CGAGATGTACAAGCGCGACCTGATCTGGGTGGTGGCCAAGATGCAGGTGATGGTGAACCGCTACCCCACC
TGGGGCGACACCGTGGAGGTGAACACCTGGGTGGCCAAGTCCGGCAAGAACGGCATGCGCCGCGACTGGC
TGATCTCCGACTGCAACACCGGCGAGATCCTGACCCGCGCCTCCTCCGTGTGGGTGATGATGAACCAGAA
GACCCGCCGCCTGTCCAAGATCCCCGACGAGGTGCGCAACGAGATCGAGCCCCACTTCGTGGACTCCGCC
CCCGTGGTGGAGGACGACGACCGCAAGCTGCCCAAGCTGGACGAGAACACCGCCGACTCCATCCGCAAGG
GCCTGACCCCCCGCTGGAACGACCTGGACGTGAACCAGCACGTGAACAACGTGAAGTACATCGGCTGGAT
CCTGGAGTCCACCCOCCCCGAGGTGCTGGAGACCCAGGAGCTGTGCTCCCTGACCCTGGAGTACCGCCGC
GAGTGCGGCCGCGAGTCCGTGCTGGAGTCCCTGACCGCCGTGGACCCCTCCGCCGAGGGCTACGCCTCCC
GCTTCCAGCACCTGCTGCGCCTGGAGGACGGCGGCGAGATCGTGAAGGCCCGCACCGAGTGGCGCCCCAA
GAACGCCGGCATCAACGGCGTGGTGCCCTCCGAGGAGTCCTCCCCCGGCGACTTCTTCTGA
SEQ ID NO: 34
Cuphea wrightii (Cw) FATB4b amino acid sequence
MVATAASSAFFPVPSADTSSSRPGKLGNGPSSLSPLKPKSIPNGGLQVKANASAPPKINGSSVGLKSGSF
KTQEDAPSAPPPRTFINQLPDWSMLLAAITTVFLAAEKQWMMLDWKPKRPDMLVDPFGLGSIVQDGLVFR
QNFSIRSYEIGADRTASIETVMNHLQETALNHVKIAGLSSDGFGRTPAMSKRDLIWVVAKMQVMVNRYPA
WGDTVEVNTWVAKSGKNGMRRDWLISDCNTGEILTRASSVWVMMNQKTRRLSKIPDEVRNEIEPHFVDSA
PVVEDDDRKLPKLDENTADSIRKGLTPRWNDLDVNQHVNNVKYIGWILESTPAEVLETQELCSLTLEYRR
ECGRESVLESLTAVDPSGEGDGSKFQHLLRLEDGGEIVKARTEWRPKNAGINGVVPSEESSPGGDFF*
SEQ ID NO: 35
Cuphea wrightii (Cw) FATB4b coding DNA sequence
TTGGTGGCTACCGCTGCAAGTTCTGCATTTTTCCCCGTACCATCCGCCGACACCTCCTCATCGAGACCCG
GAAAGCTCGGCAATGGGCCATCGAGCTTGAGCCCCCTCAAGCCGAAATCGATCCCCAATGGCGGGTTGCA
GGTTAAGGCAAACGCCAGTGCCCCTCCTAAGATCAATGGTTCCTCGGTCGGTCTGAAGTCGGGCAGTTTC
AAGACTCAGGAAGACGCTCCTTCGGCCCCTCCTCCTCGGACTTTTATCAACCAGTTGCCTGATTGGAGTA
TGCTTCTTGCTGCAATCACTACTGTCTTCTTGGCTGCAGAGAAGCAGTGGATGATGCTTGATTGGAAACC
TAAGAGGCCTGACATGCTTGTCGACCCGTTCGGATTGGGAAGTATTGTTCAGGATGGGCTTGTTTTCAGG
CAGAATTTCTCGATTAGGTCCTACGAAATAGGCGCTGATCGCACTGCGTCTATAGAGACGGTGATGAACC
ATTTGCAGGAAACAGCTCTCAATCATGTTAAGATTGCTGGGCTTTCTAGTGATGGCTTTGGTCGTACTCC
TGCGATGTCTAAACGGGACCTCATTTGGGTTGTTGCGAAAATGCAGGTCATGGTTAACCGCTACCCTGCT
TGGGGTGACACGGTTGAAGTGAATACTTGGGTTGCCAAGTCAGGGAAAAATGGTATGCGTCGTGACTGGC
TCATAAGTGATTGCAACACTGGAGAGATTCTTACAAGAGCATCAAGCGTGTGGGTCATGATGAATCAAAA
GACAAGAAGATTGTCAAAAATTCCAGATGAGGTTCGAAATGAGATAGAGCCTCATTTTGTGGACTCTGCG
CCCGTCGTTGAAGACGATGACCGGAAACTTCCCAAGCTGGATGAGAACACTGCTGACTCCATCCGCAAGG
GTCTAACTCCGAGGTGGAATGACTTGGATGTCAATCAGCACGTCAACAACGTGAAGTACATTGGGTGGAT
TCTTGAGAGTACTCCAGCAGAAGTTCTGGAGACCCAGGAATTATGTTCCCTGACCCTGGAATACAGGCGG
GAATGTGGAAGGGAGAGCGTGCTGGAGTCCCTCACTGCTGTAGATCCGTCTGGAGAGGGCGATGGGTCCA
AGTTCCAGCACCTTCTGCGGCTTGAGGATGGAGGTGAGATCGTGAAGGCGAGAACTGAGTGGCGACCAAA
GAATGCTGGAATCAATGGGGTGGTACCATCCGAGGAGTCCTCACCTGGTGGAGACTTCTTTTAA
SEQ ID NO: 36
Cuphea wrightii (Cw) FATB4b coding DNA sequence codon optimized for
Prototheca moriformis
ATGGTGGCCACCGCCGCCTCCTCCGCCTTCTTCCCCGTGCCCTCCGCCGACACCTCCTCCTCCCGCCCCG
GCAAGCTGGGCAACGGCCCCTCCTCCCTGTCCCCCCTGAAGCCCAAGTCCATCCCCAACGGCGGCCTGCA
GGTGAAGGCCAACGCCTCCGCCCCCCCCAAGATCAACGGCTCCTCCGTGGGCCTGAAGTCCGGCTCCTTC
AAGACCCAGGAGGACGCCCCCTCCGCCCCCCCCCCCCGCACCTTCATCAACCAGCTGCCCGACTGGTCCA
TGCTGCTGGCCGCCATCACCACCGTGTTCCTGGCCGCCGAGAAGCAGTGGATGATGCTGGACTGGAAGCC
CAAGCGCCCCGACATGCTGGTGGACCCCTTCGGCCTGGGCTCCATCGTGCAGGACGGCCTGGTGTTCCGC
CAGAACTTCTCCATCCGCTCCTACGAGATCGGCGCCGACCGCACCGCCTCCATCGAGACCGTGATGAACC
ACCTGCAGGAGACCGCCCTGAACCACGTGAAGATCGCCGGCCTGTCCTCCGACGGCTTCGGCCGCACCCC
CGCCATGTCCAAGCGCGACCTGATCTGGGTGGTGGCCAAGATGCAGGTGATGGTGAACCGCTACCCCGCC
TGGGGCGACACCGTGGAGGTGAACACCTGGGTGGCCAAGTCCGGCAAGAACGGCATGCGCCGCGACTGGC
TGATCTCCGACTGCAACACCGGCGAGATCCTGACCCGCGCCTCCTCCGTGTGGGTGATGATGAACCAGAA
GACCCGCCGCCTGTCCAAGATCCCCGACGAGGTGCGCAACGAGATCGAGCCCCACTTCGTGGACTCCGCC
CCCGTGGTGGAGGACGACGACCGCAAGCTGCCCAAGCTGGACGAGAACACCGCCGACTCCATCCGCAAGG
GCCTGACCCCCCGCTGGAACGACCTGGACGTGAACCAGCACGTGAACAACGTGAAGTACATCGGCTGGAT
CCTGGAGTCCACCCCCGCCGAGGTGCTGGAGACCCAGGAGCTGTGCTCCCTGACCCTGGAGTACCGCCGC
GAGTGCGGCCGCGAGTCCGTGCTGGAGTCCCTGACCGCCGTGGACCCCTCCGGCGAGGGCGACGGCTCCA
AGTTCCAGCACCTGCTGCGCCTGGAGGACGGCGGCGAGATCGTGAAGGCCCGCACCGAGTGGCGCCCCAA
GAACGCCGGCATCAACGGCGTGGTGCCCTCCGAGGAGTCCTCCCCCGGCGGCGACTTCTTCTGA
SEQ ID NO: 37
Cuphea wrightii (Cw) FATB5 amino acid sequence
MVAAAASSAFFSVPTPGTPPKPGKFGNWPSSLSVPFKPDNGGFHVKANASAHPKANGSAVNLKSGSLETP
PRSFINQLPDLSVLLSKITTVFGAAEKQWKRPGMLVEPFGVDRIFQDGVFFRQSFSIRSYEIGVDRTASI
ETLMNIFQETSLNHCKSIGLLNDGFGRTPEMCKRDLIWVVTKIQVEVNRYPTWGDTIEVNTWVSESGKNG
MGRDWLISDCRTGEILIRATSVWAMMNQNTRRLSKFPYEVRQEIAPHFVDSAPVIEDDQKLQKLDVKTGD
SIRDGLTPRWNDLDVNQHVNNVKYIGWILKSVPIEVFETQELCGVTLEYRRECGRDSVLESVTAMDPAKE
GDRCVYQHLLRLEDGADITIGRTEWRPKNAGANGAMSSGKTSNGNCLIEGRGWQPFRVVRLIF*
SEQ ID NO: 38
Cuphea wrightii (Cw) FATB5 coding DNA sequence
ATGGTGGCTGCCGCAGCAAGTTCTGCATTCTTCTCTGTTCCAACCCCGGGAACGCCCCCTAAACCCGGGA
AGTTCGGTAACTGGCCATCGAGCTTGAGCGTCCCCTTCAAGCCCGACAATGGTGGCTTTCATGTCAAGGC
AAACGCCAGTGCCCATCCTAAGGCTAATGGTTCTGCGGTAAATCTAAAGTCTGGCAGCCTCGAGACTCCT
CCTCGGAGTTTCATTAACCAGCTGCCGGACTTGAGTGTGCTTCTGTCCAAAATCACGACTGTCTTCGGGG
CGGCTGAGAAGCAGTGGAAGAGGCCCGGCATGCTCGTGGAACCGTTTGGGGTTGACAGGATTTTTCAGGA
TGGTGTTTtTTTCAGACAGAGTTTTTCTATCAGGTCTTACGAAATAGGCGTTGATCGAACAGCCTCGATA
GAGACACTGATGAACATCTTCCAGGAAACATCTTTGAATCATTGCAAGAGTATCGGTCTTCTCAACGATG
GCTTTGGTCGTACTCCTGAGATGTGTAAGAGGGACCTCATTTGGGTGGTTACGAAAATTCAGGTCGAGGT
GAATCGCTATCCTACTTGGGGTGACACTATCGAAGTCAATACTTGGGTCTCGGAGTCGGGGAAAAACGGT
ATGGGTCGGGATTGGCTGATAAGTGATTGCCGTACTGGAGAGATTCTTATAAGAGCAACGAGCGTGTGGG
CGATGATGAATCAAAACACGAGAAGATTGTCAAAATTTCCATATGAGGTTCGACAGGAGATAGCGCCTCA
TTTTGTGGACTCTGCTCCTGTCATTGAAGACGATCAAAAGTTGCAGAAGCTTGATGTGAAGACAGGTGAT
TCCATTCGCGATGGTCTAACTCCGAGATGGAATGACTTGGATGTCAATCAACACGTTAACAATGTGAAGT
ACATTGGATGGATTCTCAAGAGTGTTCCAATAGAAGTTTTCGAGACACAGGAGCTATGCGGCGTCACACT
TGAATATAGGCGGGAATGCGGAAGGGACAGTGTGCTGGAGTCAGTGACCGCTATGGATCCAGCAAAAGAG
GGAGACCGGTGTGTGTACCAGCACCTTCTTCGGCTTGAGGATGGAGCTGATATCACTATAGGCAGAACCG
AGTGGCGGCCGAAGAATGCAGGAGCCAATGGTGCAATGTCATCAGGAAAGACTTCAAATGGAAACTGTCT
CATAGAAGGAAGGGGTTGGCAACCTTTCCGAGTTGTGCGTTTAATTTTCTGA
SEQ ID NO: 39
Cuphea wrightii (Cw) FATB5 coding DNA sequence codon optimized for
Prototheca moriformis
ATGGTGGCCGCCGCCGCCTCCTCCGCCTTCTTCTCCGTGCCCACCCCCGGCACCCCCCCCAAGCCCGGCA
AGTTCGGCAACTGGCCCTCCTCCCTGTCCGTGCCCTTCAAGCCCGACAACGGCGGCTTCCACGTGAAGGC
CAACGCCTCCGCCCACCCCAAGGCCAACGGCTCCGCCGTGAACCTGAAGTCCGGCTCCCTGGAGACCCCC
CCCCGCTCCTTCATCAACCAGCTGCCCGACCTGTCCGTGCTGCTGTCCAAGATCACCACCGTGTTCGGCG
CCGCCGAGAAGCAGTGGAAGCGCCCCGGCATGCTGGTGGAGCCCTTCGGCGTGGACCGCATCTTCCAGGA
CGGCGTGTTCTTCCGCCAGTCCTTCTCCATCCGCTCCTACGAGATCGGCGTGGACCGCACCGCCTCCATC
GAGACCCTGATGAACATCTTCCAGGAGACCTCCCTGAACCACTGCAAGTCCATCGGCCTGCTGAACGACG
GCTTCGGCCQCACCCCCGAGATGTGCAAGCGCGACCTGATCTGGGTGGTGACCAAGATCCAGGTGGAGGT
GAACCGCTACCCCACCTGGGGCGACACCATCGAGGTGAACACCTGGGTGTCCGAGTCCGGCAAGAACGGC
ATGGGCCGCGACTGGCTGATCTCCGACTGCCGCACCGGUGAGATCCTGATCCGCGCCACCTCCGTGTGGG
CCATGATGAACCAGAACACCCGCCGCCTGTCCAAGTTCCCCTACGAGGTGCGCCAGGAGATCGCCCCCCA
CTTCGTGGACTCCGCCCCCGTGATCGAGGACGACCAGAAGCTGCAGAAGCTGGACGTGAAGACCGGCGAC
TCCATCCGCGACGGCCTGACCCCCCGCTGGAACGACCTGGACGTGAACCAGCACGTGAACAACGTGAAGT
ACATCGGCTGGATCCTGAAGTCCGTGCCCATCGAGGTGTTCGAGACCCAGGAGCTGTGCGGCGTGACCCT
GGAGTACCGCCGCGAGTGCGGCCGCGACTCCGTGCTGGAGTCCGTGACCGCCATGGACCCCGCCAAGGAG
GGCGACCGCTGCGTGTACCAGCACCTGCTGCGCCTGGAGGACGGCGCCGACATCACCATCGGCCGCACCG
AGTGGCGCCCCAAGAACGCCGGCGCCAACGGCGCCATGTCCTCCGGCAAGACCTCCAACGGCAACTGCCT
GATCGAGGGCCGCGGCTGGCAGCCCTTCCGCGTGGTGCGCCTGATCTTCTGA
SEQ ID NO: 40
Cuphea heterophylla (Cht) FATB1a amino acid sequence
MVAAAASSAFFSVPTPGTSTKPGNFGNWPSSLSVPFKPESNHNGGFRVKANASAHPKANGSAVNLKSGSL
ETQEDTSSSSPPPRTFIKQLPDWGMLLSKITTVFGAAERQWKRPGMLVEPFGVDRIFQDGVFFRQSFSIR
SYEIGADRTASIETLMNIFQETSLNHCKSIGLLNDGFGRTPEMCKRDLIWVVTKIQVEVNRYPTWGDTIE
VNTWVSESGKNGMGRDWLISDCRTGEILIRATSVWAMMNRKTRRLSKFPYEVRQEIAPHFVDSAPVIEDD
KKLHKLDVKTGDSIRKGLTPRWNDLDVNQHVNNVKYIGWILKSVPAEVFETQELCGVTLEYRRECGRDSV
LESVTAMDTAKEGDRSLYQHLLRLEDGADITIGRTEWRPKNAGANGAISTGKTSNENSVS*
SEQ ID NO: 41
Cuphea heterophylla (Cht) FATB1a coding DNA sequence
ATGGTGGCTGCCGCAGCAAGTTCTGCATTCTTCTCCGTTCCAACCCCGGGAACCTCCACTAAACCCGGGA
ACTTCGGCAATTGGCCATCGAGCTTGAGCGTCCCCTTCAAGCCCGAATCAAACCACAATGGTGGCTTTCG
GGTCAAGGCAAACGCCAGTGCTCATCCTAAGGCTAACGGTTCTGCAGTAAATCTAAAGTCTGGCAGCCTC
GAGACTCAGGAGGACACTTCATCGTCGTCCCCTCCTCCTCGGACTTTTATTAAGCAGTTGCCCGACTGGG
GTATGCTTCTGTCCAAAATCACGACTGTCTTCGGGGCGGCTGAGAGGCAGTGGAAGAGGCCCGGCATGCT
TGTGGAACCGTTTGGGGTTGACAGGATTTTTCAGGATGGGGTTTTTTTCAGACAGAGTTTTTCGATCAGG
TCTTACGAAATAGGCGCTGATCGAACAGCCTCAATAGAGACGCTGATGAACATCTTCCAGGAAACATCTC
TGAATCATTGTAAGAGTATCGGTCTTCTCAATGACGGCTTTGGTCGTACTCCTGAGATGTGTAAGAGGGA
CCTCATTTGGGTGGTTACGAAAATTCAGGTCGAGGTGAATCGCTATCCTACTTGGGGTGATACTATTGAG
GTCAATACTTGGGTCTCAGAGTCGGGGAAAAACGGTATGGGTCGTGATTGGCTGATAAGCGATTGCCGTA
CCGGAGAAATTCTTATAAGAGCAACGAGCGTGTGGGCTATGATGAATCGAAAGACGAGAAGATTGTCAAA
ATTTCCATATGAGGTTCGACAGGAGATAGCGCCTCATTTTGTGGACTCTGCTCCTGTCATTGAAGACGAT
AAAAAATTGCACAAGCTTGATGTTAAGACGGGTGATTCCATTCGCAAGGGTCTAACTCCAAGGTGGAATG
ACTTGGATGTCAATCAGCACGTTAACAATGTGAAGTACATTGGGTGGATTCTCAAGAGTGTTCCAGCAGA
AGTTTTCGAGACCCAGGAGCTATGCGGAGTCACCCTTGAGTACAGGCGGGAATGTGGAAGGGACAGTGTG
CTGGAGTCCGTGACCGCTATGGATACCGCAAAAGAGGGAGACCGGTCTCTGTACCAGCACCTTCTTCGGC
TTGAGGATGGGGCTGATATCACCATAGGCAGAACCGAGTGGCGGCCGAAGAATGCAGGAGCCAATGGGGC
AATATCAACAGGAAAGACTTCAAATGAAAACTCTGTCTCTTAG
SEQ ID NO: 42
Cuphea heterophylla (Cht) FATB1a coding DNA sequence codon optimized
for Prototheca moriformis
ATGGTGGCCGCCGCCGCCTCCTCCGCCTTCTTCTCCGTGCCCACCCCCGGCACCTCCACCAAGCCCGGCA
ACTTCGGCAACTGGCCCTCCTCCCTGTCCGTGCCCTTCAAGCCCGAGTCCAACCACAACGGCGGCTTCCG
CGTGAAGGCCAACGCCTCCGCCCACCCCAAGGCCAACGGCTCCGCCGTGAACCTGAAGTCCGGCTCCCTG
GAGACCCAGGAGGACACCTCCTCCTCCTCCCCCCCCCCCCGCACCTTCATCAAGCAGCTGCCCGACTGGG
GCATGCTGCTGTCCAAGATCACCACCGTGTTCGGCGCCGCCGAGCGCCAGTGGAAGCGCCCCGGCATGCT
GGTGGAGCCCTTCGGCGTGGACCGCATCTTCCAGGACGGCGTGTTCTTCCGCCAGTCCTTCTCCATCCGC
TCCTACGAGATCGGCGCCGACCGCACCGCCTCCATCGAGACCCTGATGAACATCTTCCAGGAGACCTCCC
TGAACCACTGCAAGTCCATCGGCCTGCTGAACGACGGCTTCGGCCGCACCCCCGAGATGTGCAAGCGCGA
CCTGATCTGGGTGGTGACCAAGATCCAGGTGGAGGTGAACCGCTACCCCACCTGGGGCGACACCATCGAG
GTGAACACCTGGGTGTCCGAGTCCGGCAAGAACGGCATGGGCCGCGACTGGCTGATCTCCGACTGCCGCA
CCGGCGAGATCCTGATCCGCGCCACCTCCGTGTGGGCCATGATGAACCGCAAGACCCGCCGCCTGTCCAA
GTTCCCCTACGAGGTGCGCCAGGAGATCGCCCCCCACTTCGTGGACTCCGCCCCCGTGATCGAGGACGAC
AAGAAGCTGCACAAGCTGGACGTGAAGACCGGCGACTCCATCCGCAAGGGCCTGACCCCCCGCTGGAACG
ACCTGGACGTGAACCAGCACGTGAACAACGTGAAGTACATCGGCTGGATCCTGAAGTCCGTGCCCGCCGA
GGTGTTCGAGACCCAGGAGCTGTGCGGCGTGACCCTGGAGTACCGCCGCGAGTGCGGCCGCGACTCCGTG
CTGGAGTCCGTGACCGCCATGGACACCGCCAAGGAGGGCGACCGCTCCCTGTACCAGCACCTGCTGCGCC
TGGAGGACGGCGCCGACATCACCATCGGCCGCACCGAGTGGCGCCCCAAGAACGCCGGCGCCAACGGCGC
CATCTCCACCGGCAAGACCTCCAACGAGAACTCCGTGTCCTGA
SEQ ID NO: 43
Cuphea heterophylla (Cht) FATB1b (P16S, T20P, G94S, G105W, S293F,
L305F variant) amino acid sequence
MVAAAASSAFFSVPTSGTSPKPGNFGNWPSSLSVPFKPESSHNGGFQVKANASAHPKANGSAVNLKSGSL
ETQEDTSSSSPPPRTFIKQLPDWSMLLSKITTVFWAAERQWKRPGMLVEPFGVDRIFQDGVFFRQSFSIR
SYEIGADRTASIETLMNIFQETSLNHCKSIGLLNDGFGRTPEMCKRDLIWVVTKIQVEVNRYPTWGDTIE
VNTWVSESGKNGMGRDWLISDCRTGEILIRATSVWAMMNRKTRRLSKFPYEVRQEIAPHFVDSAPVIEDD
KKLHKLDVKTGDFIRKGLTPRWNDFDVNQHVNNVKYIGWILKSVPAEVFETQELCGVTLEYRRECGRDSV
LESVTAMDTAKEGDRSLYQHLLRLEDGADITIGRTEWRPKNAGANGAISTGKTSNENSVS*
SEQ ID NO: 44
Cuphea heterophylla (Cht) FATB1b (P16S, T2OP, G94S, G105W, S293F, L305F
variant) coding DNA sequence
ATGGTGGCTGCCGCAGCAAGTTCTGCATTCTTCTCCGTTCCAACCTCGGGAACCTCCCCTAAACCCGGGA
ACTTCGGCAATTGGCCATCGAGCTTGAGCGTCCCCTTCAAGCCCGAATCAAGCCACAATGGTGGCTTTCA
GGTCAAGGCAAACGCCAGTGCCCATCCTAAGGCTAACGGTTCTGCAGTAAATCTAAAGTCTGGCAGCCTC
GAGACTCAGGAGGACACTTCATCGTCGTCCCCTCCTCCTCGGACTTTTATTAAGCAGTTGCCCGACTGGA
GTATGCTTCTGTCCAAAATCACGACTGTCTTCTGGGCGGCTGAGAGGCAGTGGAAGAGGCCCGGCATGCT
TGTGGAACCGTTTGGGGTTGACAGGATTTTTCAGGATGGGGTTTTTTTCAGACAGAGTTTTTCGATCAGG
TCTTACGAAATAGGCGCTGATCGAACAGCCTCAATAGAGACGCTGATGAACATCTTCCAGGAAACATCTC
TGAATCATTGTAAGAGTATCGGTCTTCTCAATGACGGCTTTGGTCGTACTCCTGAGATGTGTAAGAGGGA
CCTCATTTGGGTGGTTACGAAAATTCAGGTCGAGGTGAATCGCTATCCTACTTGGGGTGATACTATTGAG
GTCAATACTTGGGTCTCAGAGTCGGGGAAAAACGGTATGGGTCGTGATTGGCTGATAAGCGATTGCCGTA
CCGGAGAAATTCTTATAAGAGCAACGAGCGTGTGGGCTATGATGAATCGAAAGACGAGAAGATTGTCAAA
ATTTCCATATGAGGTTCGACAGGAGATAGCGCCTCATTTTGTGGACTCTGCTCCTGTCATTGAAGACGAT
AAAAAATTGCACAAGCTTGATGTTAAGACGGGTGATTTCATTCGCAAGGGTCTAACTCCAAGGTGGAATG
ACTTTGATGTCAATCAGCACGTTAACAATGTGAAGTACATTGGGTGGATTCTCAAGAGTGTTCCAGCAGA
AGTTTTCGAGACCCAGGAGCTATGCGGAGTCACCCTTGAGTATAGGCGGGAATGTGGAAGGGACAGTGTG
CTGGAGTCCGTGACCGCTATGGATACCGCAAAAGAGGGAGACCGGTCTCTGTACCAGCACCTTCTTCGGC
TTGAGGATGGGGCTGATATCACCATAGGCAGAACCGAGTGGCGGCCGAAGAATGCAGGAGCCAATGGGGC
AATATCAACAGGAAAGACTTCAAATGAAAACTCTGTCTCTTAG
SEQ ID NO: 45
Cuphea heterophylla (Cht) FATB1b (P16S, T20P, G94S, G105W, S293F,
L305F variant) coding DNA sequence codon optimized for Prototheca
moriformis
ATGGTGGCCGCCGCCGCCTCCTCCGCCTTCTTCTCCGTGCCCACCTCCGGCACCTCCCCCAAGCCCGGCA
ACTTCGGCAACTGGCCCTCCTCCCTGTCCGTGCCCTTCAAGCCCGAGTCCTCCCACAACGGCGGCTTCCA
GGTGAAGGCCAACGCCTCCGCCCACCCCAAGGCCAACGGCTCCGCCGTGAACCTGAAGTCCGGCTCCCTG
GAGACCCAGGAGGACACCTCCTCCTCCTCCCCCCCCCCCCGCACCTTCATCAAGCAGCTGCCCGACTGGT
CCATGCTGCTGTCCAAGATCACCACCGTGTTCTGGGCCGCCGAGCGCCAGTGGAAGCGCCCCGGCATGCT
GGTGGAGCCCTTCGGCGTGGACCGCATCTTCCAGGACGGCGTGTTCTTCCGCCAGTCCTTCTCCATCCGC
TCCTACGAGATCGGCGCCGACCGCACCGCCTCCATCGAGACCCTGATGAACATCTTCCAGGAGACCTCCC
TGAACCACTGCAAGTCCATCGGCCTGCTGAACGACGGCTTCGGCCGCACCCCCGAGATGTGCAAGCGCGA
CCTGATCTGGGTGGTGACCAAGATCCAGGTGGAGGTGAACCGCTACCCCACCTGGGGCGACACCATCGAG
GTGAACACCTGGGTGTCCGAGTCCGGCAAGAACGGCATGGGCCGCGACTGGCTGATCTCCGACTGCCGCA
CCGGCGAGATCCTGATCCGCGCCACCTCCGTGTGGGCCATGATGAACCGCAAGACCCGCCGCCTGTCCAA
GTTCCCCTACGAGGTGCGCCAGGAGATCGCCCCCCACTTCGTGGACTCCGCCCCCGTGATCGAGGACGAC
AAGAAGCTGCACAAGCTGGACGTGAAGACCGGCGACTTCATCCGCAAGGGCCTGACCCCCCGCTGGAACG
ACTTCGACGTGAACCAGCACGTGAACAACGTGAAGTACATCGGCTGGATCCTGAAGTCCGTGCCCGCCGA
GGTGTTCGAGACCCAGGAGCTGTGCGGCGTGACCCTGGAGTACCGCCGCGAGTGCGGCCGCGACTCCGTG
CTGGAGTCCGTGACCGCCATGGACACCGCCAAGGAGGGCGACCGCTCCCTGTACCAGCACCTGCTGCGCC
TGGAGGACGGCGCCGACATCACCATCGGCCGCACCGAGTGGCGCCCCAAGAACGCCGGCGCCAACGGCGC
CATCTCCACCGGCAAGACCTCCAACGAGAACTCCGTGTCCTGA
SEQ ID NO: 46
Cuphea heterophylla (Cht) FATB2b amino acid sequence
MVVAAAASSAFFPVPASGTSPKPGKFGTWLSSSSPSYKPKSNPSGGFQVKANASAHPKANGSAVSLKSGS
LNTQEGTSSSPPPRTFLNQLPDWSRLRTAITTVFVAAEKQLTMLDRKSKKPDMHVDWFGLEIIVQDGLVF
RESFSIRSYEIGADRTASIETLMNHLQDTSLNHCKSVGLLNDGFGRTPEMCKRDLIWVLTKMQIMVNRYP
TWGDTVEINSWFSQSGKIGMGRNWLISDCNTGEILIRATSIWAMMNQKTRRFSKLPNEVRQEIAPHFVDA
PPVIEDNDRKLHKFDVKTGDSICKGLTPEWNDLDVNQHVSNVKYIGWILESMPKEVLDTQELCSLTLEYR
RECGRDSVLESVTAMDPSKVGDRSQYQHLLRLEDGTDIMKGRTEWRPKNAGTNGAISTGKTSNGNSVS*
SEQ ID NO: 47
Cuphea heterophylla (Cht) FATB2b coding DNA sequence
ATGGTGGTGGCTGCTGCAGCAAGCTCTGCATTCTTCCCTGTTCCGGCATCTGGAACCTCCCCTAAACCCG
GGAAGTTCGGGACTTGGCTATCGAGCTCGAGCCCTTCCTACAAGCCCAAGTCAAACCCCAGTGGTGGATT
TCAGGTTAAGGCAAATGCCAGTGCTCATCCTAAGGCTAACGGTTCCGCAGTAAGTCTAAAGTCTGGCAGC
CTCAACACTCAGGAGGGCACTTCGTCGTCCCCTCCTCCTCGGACTTTCCTTAACCAGTTGCCTGATTGGA
GTAGGCTTCGGACTGCAATCACGACCGTCTTCGTGGCGGCAGAGAAGCAGTTGACTATGCTCGATCGAAA
GTCTAAGAAGCCTGACATGCACGTGGACTGGTTTGGGTTGGAGATTATTGTTCAGGATGGGCTCGTGTTC
AGAGAGAGTTTTTCGATCAGGTCTTACGAAATAGGCGCTGATCGAACAGCCTCTATAGAAACGTTGATGA
ACCATTTGCAGGACACATCTTTGAACCATTGTAAGAGTGTGGGTCTTCTCAATGACGGCTTTGGTCGTAC
CCCGGAGATGTGTAAAAGGGACCTCATTTGGGTGCTTACAAAAATGCAGATCATGGTGAATCGCTATCCA
ACTTGGGGCGATACTGTCGAGATCAATAGCTGGTTCTCCCAGTCCGGGAAAATCGGTATGGGTCGCAATT
GGCTAATAAGTGATTGCAACACAGGAGAAATTCTTATAAGAGCAACGAGCATTTGGGCCATGATGAATCA
AAAGACGAGAAGATTCTCAAAACTTCCAAACGAGGTTCGCCAGGAGATAGCGCCTCATTTTGTGGACGCC
CCTCCTGTCATTGAAGACAATGATCGAAAATTGCATAAGTTTGATGTGAAGACTGGTGATTCCATTTGCA
AGGGTCTAACACCGGAGTGGAATGACTTGGATGTCAATCAGCACGTAAGCAACGTGAAGTACATTGGGTG
GATTCTCGAGAGTATGCCAAAAGAAGTTTTGGACACCCAGGAGCTATGCTCTCTCACCCTTGAATATAGG
CGGGAATGCGGAAGGGATAGTGTGCTGGAGTCTGTGACCGCTATGGATCCCTCAAAAGTTGGAGACCGAT
CTCAGTACCAGCACCTTCTGCGGCTTGAAGATGGGACTGATATCATGAAGGGCAGAACTGAGTGGCGACC
AAAGAATGCAGGAACCAACGGGGCTATATCAACAGGAAAGACTTCAAATGGAAACTCGGTCTCTTAG
SEQ ID NO: 48
Cuphea heterophylla (Cht) FATB2b coding DNA sequence codon optimized
for Prototheca moriformis
ATGGTGGTGGCCGCCGCCGCCTCCTCCGCCTTCTTCCCCGTGCCCGCCTCCGGCACCTCCCCCAAGCCCG
GCAAGTTCGGCACCTGGCTGTCCTCCTCCTCCCCCTCCTACAAGCCCAAGTCCAACCCCTCCGGCGGCTT
CCAGGTGAAGGCCAACGCCTCCGCCCACCCCAAGGCCAACGGCTCCGCCGTGTCCCTGAAGTCCGGCTCC
CTGAACACCCAGGAGGGCACCTCCTCCTCCCCCCCCCCCCGCACCTTCCTGAACCAGCTGCCCGACTGGT
CCCGCCTGCGCACCGCCATCACCACCGTGTTCGTGGCCGCCGAGAAGCAGCTGACCATGCTGGACCGCAA
GTCCAAGAAGCCCGACATGCACGTGGACTGGTTCGGCCTGGAGATCATCGTGCAGGACGGCCTGGTGTTC
CGCGAGTCCTTCTCCATCCGCTCCTACGAGATCGGCGCCGACCGCACCGCCTCCATCGAGACCCTGATGA
ACCACCTGCAGGACACCTCCCTGAACCACTGCAAGTCCGTGGGCCTGCTGAACGACGGCTTCGGCCGCAC
CCCCGAGATGTGCAAGCGCGACCTGATCTGGGTGCTGACCAAGATGCAGATCATGGTGAACCGCTACCCC
ACCTGGGGCGACACCGTGGAGATCAACTCCTGGTTCTCCCAGTCCGGCAAGATCGGCATGGGCCGCAACT
GGCTGATCTCCGACTGCAACACCGGCGAGATCCTGATCCGCGCCACCTCCATCTGGGCCATGATGAACCA
GAAGACCCGCCGCTTCTCCAAGCTGCCCAACGAGGTGCGCCAGGAGATCGCCCCCCACTTCGTGGACGCC
CCCCCCGTGATCGAGGACAACGACCGCAAGCTGCACAAGTTCGACGTGAAGACCGGCGACTCCATCTGCA
AGGGCCTGACCCCCGAGTGGAACGACCTGGACGTGAACCAGCACGTGTCCAACGTGAAGTACATCGGCTG
GATCCTGGAGTCCATGCCCAAGGAGGTGCTGGACACCCAGGAGCTGTGCTCCCTGACCCTGGAGTACCGC
CGCGAGTGCGGCCGCGACTCCGTGCTGGAGTCCGTGACCGCCATGGACCCCTCCAAGGTGGGCGACCGCT
CCCAGTACCAGCACCTGCTGCGCCTGGAGGACGGCACCGACATCATGAAGGGCCGCACCGAGTGGCGCCC
CAAGAACGCCGGCACCAACGGCGCCATCTCCACCGGCAAGACCTCCAACGGCAACTCCGTGTCCTGA
SEQ ID NO: 49
Cuphea heterophylla (Cht) FATB2a (S17P, P21S, T28N, L30P, S33L, G76D,
S78P, G137W variant) amino acid sequence
MVVAAAASSAFFPVPAPGTTSKPGKFGNWPSSLSPSFKPKSNPNGGFQVKANASAHPKANGSAVSLKSGS
LNTKEDTPSSPPPRTFLNQLPDWSRLRTAITTVFVAAEKQLTMLDRKSKKPDMHVDWFGLEIIVQDWLVF
RESFSIRSYEIGADRTASIETLMNHLQDTSLNHCKSVGLLNDGFGRTPEMCKRDLIWVLTKMQIMVNRYP
TWGDTVEINSWFSQSGKIGMGRNWLISDCNTGEILIRATSIWAMMNQKTRRFSKLPNEVRQEIAPHFVDA
PPLIEDNDRKLHKFDVKTGDSICKGLTPEWNDLDVNQHVSNVKYIGWILESMPKEVLDTQELCSLTLEYR
RECGRDSVLESVTAMDPSKVGDRSQYQHLLRLEDGTDIMKGRTEWRPKNAGTNGAISTGKTSNGNSVS*
SEQ ID NO: 50
Cuphea heterophylla (Cht) FATB2a (S17P, P21S, T28N, L30P, S33L, G76D,
S78P, G137W variant) coding DNA sequence
ATGGTGGTGGCTGCTGCAGCAAGTTCTGCATTCTTCCCTGTTCCAGCACCTGGAACCACGTCTAAACCCG
GGAAGTTCGGCAATTGGCCATCGAGCTTGAGCCCTTCCTTCAAGCCCAAGTCAAACCCCAATGGTGGATT
TCAGGTTAAGGCAAATGCCAGCGCTCATCCTAAGGCTAACGGGTCTGCAGTAAGTCTAAAGTCTGGCAGC
CTCAACACTAAGGAGGACACTCCGTCGTCCCCTCCTCCTCGGACTTTCCTTAACCAGTTGCCTGATTGGA
GTAGGCTTCGGACTGCAATCACGACCGTCTTCGTGGCGGCAGAGAAGCAGTTGACTATGCTCGATCGAAA
GTCTAAGAAGCCTGACATGCACGTGGACTGGTTTGGGTTGGAGATTATTGTTCAGGATTGGCTCGTGTTC
AGAGAGAGTTTTTCGATCAGGTCTTACGAAATAGGCGCTGATCGAACAGCCTCTATAGAAACGTTGATGA
ACCATTTGCAGGACACATCTTTGAACCATTGTAAGAGTGTGGGTCTTCTCAATGACGGCTTTGGTLGTAC
CCCGGAGATGTGTAAAAGGGACCTCATTTGGGTGCTTACAAAAATGCAGATCATGGTGAATCGCTATCCA
ACTTGCCGCGATACTGTCGAGATCAATAGCTGGTTCTCCCAGTCCGGGAAAATCGGTATGGGTCGCAATT
GGCTAATAAGTGATTGCAACACAGGAGAAATTCTTATAAGAGCAACGAGCATTTGGGCCATGATGAATCA
AAAGACGAGAAGATTCTCAAAACTTCCAAACGAGGTTCGCCAGGAGATAGCTCCTCATTTTGTGGACGCC
CCTCCTCTCATTGAAGACAATGATCGAAAATTGCATAAGTTTGATGTGAAGACTGGTGATTCCATTTGCA
AGGGTCTAACACCGGAGTGGAATGACTTGGATGTCAATCAGCACGTAAGCAACGTGAAGTACATTGGGTG
GATTCTCGAGAGTATGCCAAAAGAAGTTTTGGACACCCAGGAGCTATGCTCTCTCACCCTTGAATATAGG
CGGGAATGCGGAAGGGACAGTGTGCTGGAGTCTGTGACCGCTATGGATCCCTCAAAAGTTGGAGACCGAT
CTCAGTACCAGCACCTTCTGCGGCTTGAAGATGGGACTGATATCATGAAGGGCAGAACTGAGTGGCGACC
AAAGAATGCAGGAACCAACGGGGCGATATCAACAGGAAAGACTTCAAATGGAAACTCGGTCTCTTAG
SEQ ID NO: 51
Cuphea heterophylla (Cht) FATB2a (S17P, P21S, T28N, L30P, S33L, G76D,
S78P, G137W variant) coding DNA sequence codon optimized for
Prototheca moriformis
ATGGTGGTGGCCGCCGCCGCCTCCTCCGCCTTCTTCCCCGTGCCCGCCCCCGGCACCACCTCCAAGCCCG
GCAAGTTCGGCAACTGGCCCTCCTCCCTGTCCCCCTCCTTCAAGCCCAAGTCCAACCCCAACGGCGGCTT
CCAGGTGAAGGCCAACGCCTCCGCCCACCCCAAGGCCAACGGCTCCGCCGTGTCCCTGAAGTCCGGCTCC
CTGAACACCAAGGAGGACACCCCCTCCTCCCCCCCCCCCCGCACCTTCCTGAACCAGCTGCCCGACTGGT
CCCGCCTGCGCACCGCCATCACCACCGTGTTCGTGGCCGCCGAGAAGCAGCTGACCATGCTGGACCGCAA
GTCCAAGAAGCCCGACATGCACGTGGACTGGTTCGGCCTGGAGATCATCGTGCAGGACTGGCTGGTGTTC
CGCGAGTCCTTCTCCATCCGCTCCTACGAGATCGGCGCCGACCGCACCGCCTCCATCGAGACCCTGATGA
ACCACCTGCAGGACACCTCCCTGAACCACTGCAAGTCCGTGGGCCTGCTGAACGACGGCTTCGGCCGCAC
CCCCGAGATGTGCAAGCGCGACCTGATCTGGGTGCTGACCAAGATGCAGATCATGGTGAACCGCTACCCC
ACCTGGGGCGACACCGTGGAGATCAACTCCTGGTTCTCCCAGTCCGGCAAGATCGGCATGGGCCGCAACT
GGCTGATCTCCGACTGCAACACCGGCGAGATCCTGATCCGCGCCACCTCCATCTGGGCCATGATGAACCA
GAAGACCCGCCGCTTCTCCAAGCTGCCCAACGAGGTGCGCCAGGAGATCGCCCCCCACTTCGTGGACGCC
CCCCCCCTGATCGAGGACAACGACCGCAAGCTGCACAAGTTCGACGTGAAGACCGGCGACTCCATCTGCA
AGGGCCTGACCCCCGAGTGGAACGACCTGGACGTGAACCAGCACGTGTCCAACGTGAAGTACATCGGCTG
GATCCTGGAGTCCATGCCCAAGGAGGTGCTGGACACCCAGGAGCTGTGCTCCCTGACCCTGGAGTACCGC
CGCGAGTGCGGCCGCGACTCCGTGCTGGAGTCCGTGACCGCCATGGACCCCTCCAAGGTGGGCGACCGCT
CCCAGTACCAGCACCTGCTGCGCCTGGAGGACGGCACCGACATCATGAAGGGCCGCACCGAGTGGCGCCC
CAAGAACGCCGGCACCAACGGCGCCATCTCCACCGGCAAGACCTCCAACGGCAACTCCGTGTCCTGA
SEQ ID NO: 52
Cuphea heterophylla (Cht) FATB2c (G76D, S78P variant) amino acid
sequence
MVVAAAASSAFFPVPASGTSPKPGKFGTWLSSSSPSYKPKSNPSGGFQVKANASAHPKANGSAVSLKSGS
LNTKEDTPSSPPPRTFLNQLPDWNRLRTAITTVFVAAEKQLTMLDRKSKKPDMHVDWFGLEIIVQDGLVF
RESFSIRSYEIGADRTASIETLMNHLQDTSLNHCKSVGLLNDGFGRTPEMCKRDLIWVLTKMQIMVNRYP
TWGDTVEINSWFSQSGKIGMGRNWLISDCNTGEILIRATSIWAMMNQKTRRFSKLPNEVRQEIAPHFVDA
PPVIEDNDRKLHKFDVKTGDSICKGLTPEWNDLDVNQHVSNVKYIGWILESMPKEVLDTQELCSLTLEYR
RECGRDSVLESVTAMDPSKVGDRSQYQHLLRLEDGTDIMKGRTEWRPKNAGTNGAISTGKTSNGNSVS*
SEQ ID NO: 53
Cuphea heterophylla (Cht) FATB2c (G76D, S78P variant) coding DNA
sequence
ATGGTGGTGGCTGCTGCAGCAAGCTCTGCATTCTTCCCTGTTCCGGCATCTGGAACCTCCCCTAAACCCG
GGAAGTTCGGGACTTGGCTATCGAGCTCGAGCCCTTCCTACAAGCCCAAGTCAAACCCCAGTGGTGGATT
TCAGGTTAAGGCAAATGCCAGTGCTCATCCTAAGGCTAACGGTTCCGCAGTAAGTCTAAAGTCTGGCAGC
CTCAACACTAAGGAGGACACTCCGTCGTCCCCTCCTCCTCGGACTTTCCTTAACCAGTTGCCTGATTGGA
ATAGGCTTCGGACTGCAATCACGACCGTCTTCGTGGCGGCAGAGAAGCAGTTGACTATGCTCGATCGAAA
GTCTAAGAAGCCTGACATGCACGTGGACTGGTTTGGGTTGGAGATTATTGTTCAGGATGGGCTCGTGTTC
AGAGAGAGTTTTTCGATCAGGTCTTACGAAATAGGCGCTGATCGAACAGCCTCTATAGAAACGTTGATGA
ACCATTTGCAGGACACATCTTTGAACCATTGTAAGAGTGTGGGTCTTCTCAATGACGGCTTTGGTCGTAC
CCCGGAGATGTGTAAAAGGGACCTCATTTGGGTGCTTACAAAAATGCAGATCATGGTGAATCGCTATCCA
ACTTGGGGCGATACTGTCGAGATCAATAGCTGGTTCTCCCAGTCCGGGAAAATCGGTATGGGTCGCAATT
GGCTAATAAGTGATTGCAACACAGGAGAAATTCTTATAAGAGCAACGAGCATTTGGGCCATGATGAATCA
AAAGACGAGAAGATTCTCAAAACTTCCAAACGAGGTTCGCCAGGAGATAGCGCCTCATTTTGTGGACGCC
CCTCCTGTCATTGAAGACAATGATCGAAAATTGCATAAGTTTGATGTGAAGACTGGTGATTCCATTTGCA
AGGGTCTAACACCGGAGTGGAATGACTTGGATGTCAATCAGCACGTAAGCAACGTGAAGTACATTGGGTG
GATTCTCGAGAGTATGCCAAAAGAAGTTTTGGACACCCAGGAGCTATGCTCTCTCACCCTTGAATATAGG
CGGGAATGCGGAAGGGACAGTGTGCTGGAGTCTGTGACCGCTATGGATCCCTCAAAAGTTGGGGACCGAT
CTCAGTACCAGCACCTTCTGCGGCTTGAAGATGGGACTGATATCATGAAGGGCAGAACTGAGTGGCGACC
AAAGAATGCAGGAACCAACGGGGCTATATCAACAGGAAAGACTTCAAATGGAAACTCGGTCTCTTAG
SEQ ID NO: 54
Cuphea heterophylla (Cht) FATB2c (G76D, S78P variant) coding DNA
sequence codon optimized for Prototheca moriformis
ATGGTGGTGGCCGCCGCCGCCTCCTCCGCCTTCTTCCCCGTGCCCGCCTCCGGCACCTCCCCCAAGCCCG
GCAAGTTCGGCACCTGGCTGTCCTCCTCCTCCCCCTCCTACAAGCCCAAGTCCAACCCCTCCGGCGGCTT
CCAGGTGAAGGCCAACGCCTCCGCCCACCCCAAGGCCAACGGCTCCGCCGTGTCCCTGAAGTCCGGCTCC
CTGAACACCAAGGAGGACACCCCCTCCTCCCCCCCCCCCCGCACCTTCCTGAACCAGCTGCCCGACTGGA
ACCGCCTGCGCACCGCCATCACCACCGTGTTCGTGGCCGCCGAGAAGCAGCTGACCATGCTGGACCGCAA
GTCCAAGAAGCCCGACATGCACGTGGACTGGTTCGGCCTGGAGATCATCGTGCAGGACGGCCTGGTGTTC
CGCGAGTCCTTCTCCATCCGCTCCTACGAGATCGGCGCCGACCGCACCGCCTCCATCGAGACCCTGATGA
ACCACCTGCAGGACACCTCCCTGAACCACTGCAAGTCCGTGGGCCTGCTGAACGACGGCTTCGGCCGCAC
CCCCGAGATGTGCAAGCGCGACCTGATCTGGGTGCTGACCAAGATGCAGATCATGGTGAACCGCTACCCC
ACCTGGGGCGACACCGTGGAGATCAACTCCTGGTTCTCCCAGTCCGGCAAGATCGGCATGGGCCGCAACT
GGCTGATCTCCGACTGCAACACCGGCGAGATCCTGATCCGCGCCACCTCCATCTGGGCCATGATGAACCA
GAAGACCCGCCGCTTCTCCAAGCTGCCCAACGAGGTGCGCCAGGAGATCGCCCCCCACTTCGTGGACGCC
CCCCCCGTGATCGAGGACAACGACCGCAAGCTGCACAAGTTCGACGTGAAGACCGGCGACTCCATCTGCA
AGGGCCTGACCCCCGAGTGGAACGACCTGGACGTGAACCAGCACGTGTCCAACGTGAAGTACATCGGCTG
GATCCTGGAGTCCATGCCCAAGGAGGTGCTGGACACCCAGGAGCTGTGCTCCCTGACCCTGGAGTACCGC
CGCGAGTGCGGCCGCGACTCCGTGCTGGAGTCCGTGACCGCCATGGACCCCTCCAAGGTGGGCGACCGCT
CCCAGTACCAGCACCTGCTGCGCCTGGAGGACGGCACCGACATCATGAAGGGCCGCACCGAGTGGCGCCC
CAAGAACGCCGGCACCAACGGCGCCATCTCCACCGGCAAGACCTCCAACGGCAACTCCGTGTCCTGA
SEQ ID NO: 55
Cuphea heterophylla (Cht) FATB2d (S21P, T28N, L30P, S33L, G76D, R97L,
H124L, W127L, I132S, K258N, C303R, E309G, K334T, T386A variant) amino
acid sequence
MVVAAAASSAFFPVPAPGTTSKPGKFGNWPSSLSPSFKPKSNPNGGFQVKANASAHPKANGSAVSLKSGS
LNTQEDTSSSPPPRTFLNQLPDWSRLLTAISTVFVAAEKQLTMLDRKSKRPDMLVDLFGLESIVQDGLVF
RESYSIRSYEIGADRTASIETLMNHLQDTSLNHCKSVGLLNDGFGRTPEMCKRDLIWVLTKMQIMVNRYP
TWGDTVEINSWFSQSGKIGMGRNWLISDCNTGEILIRATSIWAMMNQNTRRFSKLPNEVRQEIAPHFVDA
PPVIEDNDRKLHKFDVKTGDSIRKGLTPGWNDLDVNQHVSNVKYIGWILESMPTEVLETQELCSLTLEYR
RECGRESVLESVTAMNPSKVGDRSQYQHLLRLEDGADIMKGRTEWRPKNAGTNGAISTGKTSNGNSVS*
SEQ ID NO: 56
Cuphea heterophylla (Cht) FATB2d (S21P, T28N, L30P, S33L, G76D, R97L,
H124L, W127L, I132S, K258N, C303R, E309G, K334T, T386A variant) coding
DNA sequence
ATGGTGGTGGCTGCTGCAGCAAGTTCTGCATTCTTCCCTGTTCCAGCACCTGGAACCACGTCTAAACCCG
GGAAGTTCGGCAATTGGCCATCGAGCTTGAGCCCTTCCTTCAAGCCCAAGTCAAACCCCAATGGTGGATT
TCAGGTTAAGGCAAATGCCAGTGCTCATCCTAAGGCTAACGGTTCTGCGGTAAGTCTAAAGTCTGGCAGC
CTCAACACTCAGGAGGACACTTCGTCGTCCCCTCCTCCTCGGACATTCCTTAACCAGTTGCCTGATTGGA
GTAGGCTTCTGACTGCAATCTCGACCGTCTTCGTGGCGGCAGAGAAGCAGTTGACTATGCTCGATCGAAA
ATCTAAGAGGCCTGACATGCTCGTGGACTTGTTTGGGTTGGAGAGTATTGTTCAGGATGGGCTCGTGTTC
AGAGAGAGTTATTCGATCAGGTCTTACGAAATAGGCGCTGATCGAACAGCCTCTATAGAAACGTTGATGA
ACCATTTGCAGGACACATCTTTGAACCATTGTAAGAGTGTGGGTCTTCTCAATGACGGCTTTGGTCGTAC
CCCGGAGATGTGTAAAAGGGACCTCATTTGGGTGCTTACAAAAATGCAGATCATGGTGAATCGCTATCCA
ACTTGGGGCGATACTGTCGAGATCAATAGCTGGTTCTCCCAGTCCGGGAAAATCGGTATGGGTCGCAATT
GGCTAATAAGTGATTGCAACACAGGAGAAATTCTTATAAGAGCAACGAGCATTTGGGCCATGATGAATCA
AAATACGAGAAGATTCTCAAAACTTCCAAACGAGGTTCGCCAGGAGATAGCGCCTCATTTTGTTGACGCT
CCTCCTGTCATTGAAGACAATGATCGAAAATTGCATAAGTTTGATGTGAAGACTGGTGATTCCATTCGCA
AGGGTCTAACTCCGGGGTGGAATGACTTGGATGTCAATCAGCACGTAAGCAACGTGAAGTACATTGGGTG
GATTCTCGAGAGTATGCCAACAGAAGTTTTGGAGACCCAGGAGCTATGCTCTCTCACCCTTGAATATAGG
CGGGAATGCGGAAGGGAAAGTGTGCTGGAGTCCGTGACCGCTATGAATCCCTCAAAAGTTGGAGACCGGT
CTCAGTACCAGCACCTTCTACGGCTTGAGGATGGGGCTGATATCATGAAGGGCAGAACTGAGTGGCGACC
AAAGAATGCAGGAACCAACGGGGCGATATCAACAGGAAAGACTTCAAATGGAAACTCGGTCTCTTAG
SEQ ID NO: 57
Cuphea heterophylla (Cht) FATB2d (S21P, T28N, L30P, S33L, G76D, R97L,
H124L, W127L, I132S, K258N, C303R, E309G, K334T, T386A variant) coding
DNA sequence codon optimized for Prototheca moriformis
ATGGTGGTGGCCGCCGCCGCCTCCTCCGCCTTCTTCCCCGTGCCCGCCCCCGGCACCACCTCCAAGCCCG
GCAAGTTCGGCAACTGGCCCTCCTCCCTGTCCCCCTCCTTCAAGCCCAAGTCCAACCCCAACGGCGGCTT
CCAGGTGAAGGCCAACGCCTCCGCCCACCCCAAGGCCAACGGCTCCGCCGTGTCCCTGAAGTCCGGCTCC
CTGAACACCCAGGAGGACACCTCCTCCTCCCCCCCCCCCCGCACCTTCCTGAACCAGCTGCCCGACTGGT
CCCGCCTGCTGACCGCCATCTCCACCGTGTTCGTGGCCGCCGAGAAGCAGCTGACCATGCTGGACCGCAA
GTCCAAGCGCCCCGACATGCTGGTGGACCTGTTCGGCCTGGAGTCCATCGTGCAGGACGGCCTGGTGTTC
CGCGAGTCCTACTCCATCCGCTCCTACGAGATCGGCGCCGACCGCACCGCCTCCATCGAGACCCTGATGA
ACCACCTGCAGGACACCTCCCTGAACCACTGCAAGTCCGTGGGCCTGCTGAACGACGGCTTCGGCCGCAC
CCCCGAGATGTGCAAGCGCGACCTGATCTGGGTGCTGACCAAGATGCAGATCATGGTGAACCGCTACCCC
ACCTGGGGCGACACCGTGGAGATCAACTCCTGGTTCTCCCAGTCCGGCAAGATCGGCATGGGCCGCAACT
GGCTGATCTCCGACtGCAACACCGGCGAGATCCTGATCCGCGCCACCTCCATCTGGGCCATGATGAACCA
GAACACCCGCCGCTTCTCCAAGCTGCCCAACGAGGTGCGCCAGGAGATCGCCCCCCACTTCGTGGACGCC
CCCCCCGTGATCGAGGACAACGACCGCAAGCTGCACAAGTTCGACGTGAAGACCGGCGACTCCATCCGCA
AGGGCCTGACCCCCGGCTGGAACGACCTGGACGTGAACCAGCACGTGTCCAACGTGAAGTACATCGGCTG
GATCCTGGAGTCCATGCCCACCGAGGTGCTGGAGACCCAGGAGCTGTGCTCCCTGACCCTGGAGTACCGC
CGCGAGTGCGGCCGCGAGTCCGTGCTGGAGTCCGTGACCGCCATGAACCCCTCCAAGGTGGGCGACCGCT
CCCAGTACCAGCACCTGCTGCGCCTGGAGGACGGCGCCGACATCATGAAGGGCCGCACCGAGTGGCGCCC
CAAGAACGCCGGCACCAACGGCGCCATCTCCACCGGCAAGACCTCCAACGGCAACTCCGTGTCCTGA
SEQ ID NO: 58
Cuphea heterophylla (Cht) FATB2e (G76D, R97L, H124L, I132S, G152S,
H165L, T211N, K258N, C303R, E309G, K334T, T386A variant) amino acid
sequence
MVVAAAASSAFFPVPASGTSPKPGKFGTWLSSSSPSYKPKSNPSGGFQVKANASAHPKANGSAVSLKSGS
LNTQEDTSSSPPPQTFLNQLPDWSRLLTAISTVFVAAEKQLTMLDRKSKRPDMLVDWFGLESIVQDGLVF
RESYSIRSYEISADRTASIETVMNLLQETSLNHCKSMGILNDGFGRTPEMCKRDLIWVLTKMQILVNRYP
NWGDTVEINSWFSQSGKIGMGRNWLISDCNTGEILIRATSIWAMMNQNTRRFSKLPNEVRQEIAPHFVDA
PPVIEDNDRKLHKFDVKTGDSIRKGLTPGWNDLDVNQHVSNVKYIGWILESMPTEVLETQELCSLTLEYR
RECGRDSVLESVTAMNPSKVGDRSQYQHLLRLEDGADIMKGRTEWRPKNAGTNGAISTGKTSNGNSVS*
SEQ ID NO: 59
Cuphea heterophylla (Cht) FATB2e (G76D, R97L, H124L, I132S, G152S,
H165L, T211N, K258N, C303R, E309G, K334T, T386A variant) coding DNA
sequence
ATGGTGGTGGCTGCTGCAGCAAGCTCTGCATTCTTCCCTGTTCCGGCATCTGGAACCTCCCCTAAACCCG
GGAAGTTCGGGACTTGGCTATCGAGCTCGAGCCCTTCCTACAAGCCCAAGTCAAACCCCAGTGGTGGATT
TCAGGTTAAGGCAAATGCCAGTGCTCATCCTAAGGCTAACGGTTCTGCAGTAAGTCTAAAGTCTGGCAGC
CTCAACACTCAGGAGGACACTTCGTCGTCCCCTCCTCCTCAGACATTCCTTAACCAGTTGCCTGATTGGA
GTAGGCTTCTGACAGCAATCTCGACCGTCTTCGTGGCGGCAGAGAAGCAGTTGACTATGCTCGATCGAAA
ATCTAAAAGGCCTGACATGCTCGTGGACTGGTTTGGGTTGGAGAGTATTGTTCAGGATGGGCTCGTGTTC
AGAGAGAGTTATTCGATCAGGTCTTACGAAATAAGCGCTGATCGAACAGCCTCTATAGAGACGGTGATGA
ACCTCTTGCAGGAAACATCTCTCAATCATTGTAAGAGTATGGGTATTCTCAATGACGGCTTTGGTCGTAC
CCCGGAGATGTGCAAAAGGGACCTCATTTGGGTGCTTACAAAAATGCAGATCTTGGTGAATCGCTATCCA
AATTGGGGTGATACTGTCGAGATCAATAGCTGGTTCTCCCAGTCCGGGAAAATCGGTATGGGTCGCAATT
GGCTAATAAGTGATTGCAACACAGGAGAAATTCTTATAAGAGCAACGAGCATTTGGGCCATGATGAATCA
AAATACGAGAAGATTCTCAAAACTTCCAAACGAGGTTCGCCAGGAGATAGCGCCTCATTTTGTTGACGCT
CCTCCTGTCATTGAAGACAATGATCGAAAATTGCATAAGTTTGATGTGAAGACTGGTGATTCCATTCGCA
AGGGTCTAACTCCGGGGTGGAATGACTTGGATGTCAATCAGCACGTAAGCAACGTGAAGTACATTGGGTG
GATTCTCGAGAGTATGCCAACAGAAGTTTTGGAGACCCAGGAGCTATGCTCTCTCACCCTTGAATATAGG
CGGGAATGCGGAAGGGACAGTGTGCTGGAGTCCGTGACCGCTATGAATCCCTCAAAAGTTGGAGACCGGT
CTCAGTACCAGCACCTTCTACGGCTTGAGGATGGGGCTGATATCATGAAGGGCAGAACTGAGTGGCGACC
AAAGAATGCAGGAACCAACGGGGCGATATCAACAGGAAAGACTTCAAATGGAAACTCGGTCTCTTAG
SEQ ID NO: 60
Cuphea heterophylla (Cht) FATB2e (G76D, R97L, H124L, I132S, G152S,
H165L, T211N, K258N, C303R, E309G, K334T, T386A variant) coding DNA
sequence codon optimized for Prototheca moriformis
ATGGTGGTGGCCGCCGCCGCCTCCTCCGCCTTCTTCCCCGTGCCCGCCTCCGGCACCTCCCCCAAGCCCG
GCAAGTTCGGCACCTGGCTGTCCTCCTCCTCCCCCTCCTACAAGCCCAAGTCCAACCCCTCCGGCGGCTT
CCAGGTGAAGGCCAACGCCTCCGCCCACCCCAAGGCCAACGGCTCCGCCGTGTCCCTGAAGTCCGGCTCC
CTGAACACCCAGGAGGACACCTCCTCCTCCCCCCCCCCCCAGACCTTCCTGAACCAGCTGCCCGACTGGT
CCCGCCTGCTGACCGCCATCTCCACCGTGTTCGTGGCCGCCGAGAAGCAGCTGACCATGCTGGACCGCAA
GTCCAAGCGCCCCGACATGCTGGTGGACTGGTTCGGCCTGGAGTCCATCGTGCAGGACGGCCTGGTGTTC
CGCGAGTCCTACTCCATCCGCTCCTACGAGATCTCCGCCGACCGCACCGCCTCCATCGAGACCGTGATGA
ACCTGCTGCAGGAGACCTCCCTGAACCACTGCAAGTCCATGGGCATCCTGAACGACGGCTTCGGCCGCAC
CCCCGAGATGTGCAAGCGCGACCTGATCTGGGTGCTGACCAAGATGCAGATCCTGGTGAACCGCTACCCC
AACTGGGGCGACACCGTGGAGATCAACTCCTGGTTCTCCCAGTCCGGCAAGATCGGCATGGGCCGCAACT
GGCTGATCTCCGACTGCAACACCGGCGAGATCCTGATCCGCGCCACCTCCATCTGGGCCATGATGAACCA
GAACACCCGCCGCTTCTCCAAGCTGCCCAACGAGGTGCGCCAGGAGATCGCCCCCCACTTCGTGGACGCC
CCCCCCGTGATCGAGGACAACGACCGCAAGCTGCACAAGTTCGACGTGAAGACCGGCGACTCCATCCGCA
AGGGCCTGACCCCCGGCTGGAACGACCTGGACGTGAACCAGCACGTGTCCAACGTGAAGTACATCGGCTG
GATCCTGGAGTCCATGCCCACCGAGGTGCTGGAGACCCAGGAGCTGTGCTCCCTGACCCTGGAGTACCGC
CGCGAGTGCGGCCGCGACTCCGTGCTGGAGTCCGTGACCGCCATGAACCCCTCCAAGGTGGGCGACCGCT
CCCAGTACCAGCACCTGCTGCGCCTGGAGGACGGCGCCGACATCATGAAGGGCCGCACCGAGTGGCGCCC
CAAGAACGCCGGCACCAACGGCGCCATCTCCACCGGCAAGACCTCCAACGGCAACTCCGTGTCCTGA
SEQ ID NO: 61
Cuphea heterophylla (Cht) FATB2f (R97L, H124L, I132S, G152S, H165L,
T211N variant) amino acid sequence
MVVAAAASSAFFPVPASGTSPKPGKFGTWLSSSSPSYKPKSNPSGGFQVKANASAHPKANGSAVSLKSGS
LNTQEGTSSSPPPRTFLNQLPDWSRLLTAISTVFVAAEKQLTMLDRKSKRPDMLVDWFGLESIVQDGLVF
RESYSIRSYEISADRTASIETVMNLLQETSLNHCKSMGILNDGFGRTPEMCKRDLIWVLTKMQILVNRYP
NWGDTVEINSWFSQSGKIGMGRNWLISDCNTGEILIRATSIWAMMNQKTRRFSKLPNEVRQEIAPHFVDA
PPVIEDNDRKLHKFDVKTGDSICKGLTPEWNDLDVNQHVSNVKYIGWILESMPKEVLDTQELCSLTLEYR
RECGRDSVLESVTAMDPSKVGDRSQYQHLLRLEDGTDIMKGRTEWRPKNAGTNGAISTGKTSNGNSVS*
SEQ ID NO: 62
Cuphea heterophylla (Cht) FATB2f (R97L, H124L, I132S, G152S, H165L,
T211N variant) coding DNA sequence
ATGGTGGTGGCTGCTGCAGCAAGCTCTGCATTCTTCCCTGTTCCGGCATCTGGAACCTCCCCTAAACCCG
GGAAGTTCGGGACTTGGCTATCGAGCTCGAGCCCTTCCTACAAGCCCAAGTCAAACCCCAGTGGTGGATT
TCAGGTTAAAGCAAATGCCAGTGCTCATCCTAAGGCTAACGGTTCCGCAGTAAGTCTAAAGTCTGGCAGC
CTCAACACTCAGGAGGGCACTTCGTCGTCCCCTCCTCCTCGGACATTCCTTAACCAGTTGCCTGATTGGA
GTAGGCTTCTGACTGCAATCTCGACCGTCTTCGTGGCGGCAGAGAAGCAGTTGACTATGCTCGATCGAAA
ATCTAAGAGGCCTGACATGCTCGTGGACTGGTTTGGGTTGGAGAGTATTGTTCAGGATGGGCTCGTGTTC
AGAGAGAGTTATTCGATCAGGTCTTACGAAATAAGCGCTGATCGAACAGCCTCTATAGAGACGGTGATGA
ACCTCTTGCAGGAAACATCTCTCAATCATTGTAAGAGTATGGGTATTCTCAATGACGGCTTTGGTCGTAC
CCCGGAGATGTGCAAAAGGGACCTCATTTGGGTGCTTACAAAAATGCAGATCTTGGTGAATCGCTATCCA
AATTGGGGTGATACTGTCGAGATCAATAGCTGGTTCTCCCAGTCCGGGAAAATCGGTATGGGTCGCAATT
GGCTAATAAGTGATTGCAACACAGGAGAAATTCTTATAAGAGCAACGAGCATTTGGGCCATGATGAATCA
AAAGACGAGAAGATTCTCAAAACTTCCAAATGAGGTTCGCCAGGAGATAGCGCCTCATTTTGTGGACGCC
CCTCCTGTCATTGAAGACAATGATCGAAAATTGCATAAGTTTGATGTGAAGACTGGTGATTCCATTTGCA
AGGGTCTAACACCGGAGTGGAACGACTTGGATGTCAATCAGCACGTAAGCAACGTGAAGTACATTGGGTG
GATTCTCGAGAGTATGCCAAAAGAAGTTTTGGACACCCAGGAGCTATGCTCTCTCACCCTTGAATATAGG
CGGGAATGCGGAAGGGACAGTGTGCTGGAGTCTGTGACCGCTATGGATCCCTCAAAAGTTGGAGACCGAT
CTCAGTACCAGCACCTTCTGCGGCTTGAAGATGGGACTGATATCATGAAGGGCAGAACTGAGTGGCGACC
AAAGAATGCAGGAACCAACGGGGCGATATCAACAGGAAAGACTTCAAATGGAAACTCGGTCTCTTAG
SEQ ID NO: 63
Cuphea heterophylla (Cht) FATB2f (R97L, H124L, I132S, G152S, H165L,
T211N variant) coding DNA sequence codon optimized for Prototheca
moriformis
ATGGTGGTGGCCGCCGCCGCCTCCTCCGCCTTCTTCCCCGTGCCCGCCTCCGGCACCTCCCCCAAGCCCG
GCAAGTTCGGCACCTGGCTGTCCTCCTCCTCCCCCTCCTACAAGCCCAAGTCCAACCCCTCCGGCGGCTT
CCAGGTGAAGGCCAACGCCTCCGCCCACCCCAAGGCCAACGGCTCCGCCGTGTCCCTGAAGTCCGGCTCC
CTGAACACCCAGGAGGGCACCTCCTCCTCCCCCCCCCCCCGCACCTTCCTGAACCAGCTGCCCGACTGGT
CCCGCCTGCTGACCGCCATCTCCACCGTGTTCGTGGCCGCCGAGAAGCAGCTGACCATGCTGGACCGCAA
GTCCAAGCGCCCCGACATGCTGGTGGACTGGTTCGGCCTGGAGTCCATCGTGCAGGACGGCCTGGTGTTC
CGCGAGTCCTACTCCATCCGCTCCTACGAGATCTCCGCCGACCGCACCGCCTCCATCGAGACCGTGATGA
ACCTGCTGCAGGAGACCTCCCTGAACCACTGCAAGTCCATGGGCATCCTGAACGACGGCTTCGGCCGCAC
CCCCGAGATGTGCAAGCGCGACCTGATCTGGGTGCTGACCAAGATGCAGATCCTGGTGAACCGCTACCCC
AACTGGGGCGACACCGTGGAGATCAACTCCTGGTTCTCCCAGTCCGGCAAGATCGGCATGGGCCGCAACT
GGCTGATCTCCGACTGCAACACCGGCGAGATCCTGATCCGCGCCACCTCCATCTGGGCCATGATGAACCA
GAAGACCCGCCGCTTCTCCAAGCTGCCCAACGAGGTGCGCCAGGAGATCGCCCCCCACTTCGTGGACGCC
CCCCCCGTGATCGAGGACAACGACCGCAAGCTGCACAAGTTCGACGTGAAGACCGGCGACTCCATCTGCA
AGGGCCTGACCCCCGAGTGGAACGACCTGGACGTGAACCAGCACGTGTCCAACGTGAAGTACATCGGCTG
GATCCTGGAGTCCATGCCCAAGGAGGTGCTGGACACCCAGGAGCTGTGCTCCCTGACCCTGGAGTACCGC
CGCGAGTGCGGCCGCGACTCCGTGCTGGAGTCCGTGACCGCCATGGACCCCTCCAAGGTGGGCGACCGCT
CCCAGTACCAGCACCTGCTGCGCCTGGAGGACGGCACCGACATCATGAAGGGCCGCACCGAGTGGCGCCC
CAAGAACGCCGGCACCAACGGCGCCATCTCCACCGGCAAGACCTCCAACGGCAACTCCGTGTCCTGA
SEQ ID NO: 64
Cuphea heterophylla (Cht) FATB2g (A6T, A16V, S17P, G76D, R97L, H124L,
I132S, S1431, G152S, A157T, H165L, T211N, G414A variant) amino acid
sequence
MVVAATASSAFFPVPVPGTSPKPGKFGTWLSSSSPSYKPKSNPSGGFQVKANASAHPKANGSAVSLKSGS
LNTQEDTSSSPPPRTFLNQLPDWSRLLTAISTVFVAAEKQLTMLDRKSKRPDMLVDWFGLESIVQDGLVF
REIYSIRSYEISADRTTSIETVMNLLQETSLNHCKSMGILNDGFGRTPEMCKRDLIWVLTKMQILVNRYP
NWGDTVEINSWFSQSGKIGMGRNWLISDCNTGEILIRATSIWAMMNQKTRRFSKLPNEVRQEIAPHFVDA
PPVIEDNDRKLHKFDVKTGDSICKGLTPEWNDLDVNQHVSNVKYIGWILESMPKEVLDTQELCSLTLEYR
RECGRDSVLESVTAMDPSKVGDRSQYQHLLRLEDGTDIMKGRTEWRPKNAGTNGAISTGKTSNANSVS*
SEQ ID NO: 65
Cuphea heterophylla (Cht) FATB2g (A6T, A16V, S17P, G76D, R97L, H124L,
I132S, S1431, G152S, A157T, H165L, T211N, G414A variant) coding DNA
sequence
ATGGTGGTGGCTGCTACAGCAAGTTCTGCATTCTTCCCTGTTCCTGTACCTGGAACCTCCCCTAAACCCG
GAAAGTTCGGGACTTGGCTATCGAGCTCGAGCCCTTCCTACAAGCCCAAGTCAAACCCCAGTGGTGGATT
TCAGGTTAAGGCAAATGCCAGTGCTCATCCTAAGGCTAACGGTTCTGCAGTAAGTCTAAAGTCTGGCAGC
CTCAACACTCAGGAGGACACTTCGTCGTCCCCTCCTCCTCGGACATTCCTTAACCAGTTGCCTGATTGGA
GTAGGCTTCTGACTGCAATCTCGACCGTCTTCGTGGCGGCAGAGAAGCAGTTGACTATGCTCGATCGAAA
ATCTAAGAGGCCTGACATGCTCGTGGACTGGTTTGGGTTGGAGAGTATTGTTCAGGATGGGCTCGTGTTC
AGAGAGATTTATTCGATCAGGTCTTACGAAATAAGCGCTGATCGAACAACCTCTATAGAGACGGTGATGA
ACCTCTTGCAGGAAACATCTCTCAATCATTGTAAGAGTATGGGTATTCTCAATGACGGCTTTGGTCGTAC
CCCGGAGATGTGCAAAAGGGACCTCATTTGGGTGCTTACAAAAATGCAGATCTTGGTGAATCGCTATCCA
AATTGGGGTGATACTGTCGAGATCAATAGCTGGTTCTCCCAGTCCGGGAAAATCGGTATGGGTCGCAATT
GGCTAATAAGTGATTGCAACACAGGAGAAATTCTTATAAGAGCAACGAGCATTTGGGCCATGATGAATCA
AAAGACGAGAAGATTCTCAAAACTTCCAAACGAGGTTCGCCAGGAGATAGCGCCTCATTTTGTGGACGCC
CCTCCTGTCATTGAAGACAATGATCGAAAATTGCATAAGTTTGATGTGAAGACTGGTGATTCCATTTGCA
AGGGTCTAACACCGGAGTGGAATGACTTGGATGTCAATCAGCACGTAAGCAACGTGAAGTACATTGGGTG
GATTCTCGAGAGTATGCCAAAAGAAGTTTTGGACACCCAGGAGCTATGCTCTCTCACCCTTGAATATAGG
CGGGAATGCGGAAGGGACAGTGTGCTGGAGTCTGTGACCGCTATGGATCCCTCAAAAGTTGGAGACCGAT
CTCAGTACCAGCACCTTCTGCGGCTTGAAGATGGGACTGATATCATGAAGGGCAGAACTGAGTGGCGACC
AAAGAATGCAGGAACCAACGGGGCGATATCAACAGGAAAGACTTCAAATGCAAACTCGGTCTCTTAG
SEQ ID NO: 66
Cuphea heterophylla (Cht) FATB2g (A6T, A16V, S17P, G76D, R97L, H124L,
I132S, S1431, G152S, A157T, H165L, T211N, G414A variant) coding DNA
sequence codon optimized for Prototheca moriformis
ATGGTGGTGGCCGCCACCGCCTCCTCCGCCTTCTTCCCCGTGCCCGTGCCCGGCACCTCCCCCAAGCCCG
GCAAGTTCGGCACCTGGCTGTCCTCCTCCTCCCCCTCCTACAAGCCCAAGTCCAACCCCTCCGGCGGCTT
CCAGGTGAAGGCCAACGCCTCCGCCCACCCCAAGGCCAACGGCTCCGCCGTGTCCCTGAAGTCCGGCTCC
CTGAACACCCAGGAGGACACCTCCTCCTCCCCCCCCCCCCGCACCTTCCTGAACCAGCTGCCCGACTGGT
CCCGCCTGCTGACCGCCATCTCCACCGTGTTCGTGGCCGCCGAGAAGCAGCTGACCATGCTGGACCGCAA
GTCCAAGCGCCCCGACATGCTGGTGGACTGGTTCGGCCTGGAGTCCATCGTGCAGGACGGCCTGGTGTTC
CGCGAGATCTACTCCATCCGCTCCTACGAGATCTCCGCCGACCGCACCACCTCCATCGAGACCGTGATGA
ACCTGCTGCAGGAGACCTCCCTGAACCACTGCAAGTCCATGGGCATCCTGAACGACGGCTTCGGCCGCAC
CCCCGAGATGTGCAAGCGCGACCTGATCTGGGTGCTGACCAAGATGCAGATCCTGGTGAACCGCTACCCC
AACTGGGGCGACACCGTGGAGATCAACTCCTGGTTCTCCCAGTCCGGCAAGATCGGCATGGGCCGCAACT
GGCTGATCTCCGACTGCAACACCGGCGAGATCCTGATCCGCGCCACCTCCATCTGGGCCATGATGAACCA
GAAGACCCGCCGCTTCTCCAAGCTGCCCAACGAGGTGCGCCAGGAGATCGCCCCCCACTTCGTGGACGCC
CCCCCCGTGATCGAGGACAACGACCGCAAGCTGCACAAGTTCGACGTGAAGACCGGCGACTCCATCTGCA
AGGGCCTGACCCCCGAGTGGAACGACCTGGACGTGAACCAGCACGTGTCCAACGTGAAGTACATCGGCTG
GATCCTGGAGTCCATGCCCAAGGAGGTGCTGGACACCCAGGAGCTGTGCTCCCTGACCCTGGAGTACCGC
CGCGACTGCGGCCGCGACTCCGTGCTGGAGTCCGTGACCGCCATGGACCCCTCCAAGGTGGGCGACCGCT
CCCAGTACCAGCACCTGCTGCGCCTGGAGGACGGCACCGACATCATGAAGGGCCGCACCGAGTGGCGCCC
CAAGAACGCCGGCACCAACGGCGCCATCTCCACCGGCAAGACCTCCAACGCCAACTCCGTGTCCTGA
SEQ ID NO: 67
Cuphea heterophylla (Cht) FATB3aamino acid sequence
MVATAASSAFFPVPSPDTSSRPGKLGNGSSSLRPLKPKFVANAGLQVKANASAPPKINGSSVSLKSCSLK
THEDTPSAPPPRTFINQLPDWSMLLAAITTVFLAAEKQWMMLDWKPKRPDMLVDPFGLGRIVQDGLVFRQ
NFSIRSYEIGADRTASIETVMNHLQETALNHVKSAGLLNEGFGRTPEMYKRDLIWVVAKMQVMVNRYPTW
GDTVEVNTWVAKSGKNGMRRDWLISDCNTGEILTRASSVWVMMNQKTRKLSKIPDEVRHEIEPHFVDSAP
VIEDDDWKLPKLDEKTADSIRKGLTPKWNDLDVNQHVNNVKYIGWILESTPPEVLETQELCSLTLEYRRE
CGRESVLESLTAVDPSGKGFGPQFQHLLRLEDGGEIVKGRTEWRPKTAGINGTIASGETSPGNS*
SEQ ID NO: 68
Cuphea heterophylla (Cht) FATB3a coding DNA sequence
ATGGTGGCCACCGCTGCAAGTTCTGCATTCTTCCCGGTGCCGTCCCCGGACACCTCCTCTAGACCGGGAA
AGCTCGGAAATGGGTCATCAAGCTTGAGGCCCCTCAAGCCCAAATTTGTTGCCAATGCTGGGCTGCAGGT
TAAGGCAAACGCCAGTGCCCCTCCTAAGATCAATGGTTCCTCGGTCAGTCTAAAGTCTTGCAGTCTCAAG
ACTCATGAAGACACTCCTTCAGCTCCTCCTCCGCGGACTTTTATCAACCAGTTGCCTGATTGGAGCATGC
TTCTTGCTGCAATCACTACTGTCTTCTTGGCAGCAGAGAAGCAGTGGATGATGCTTGATTGGAAACCAAA
GAGGCCTGACATGCTTGTGGACCCGTTCGGATTGGGAAGGATTGTTCAGGATGGGCTTGTGTTCAGGCAG
AATTTTTCGATTAGGTCCTATGAAATAGGCGCTGATCGCACTGCATCCATAGAGACGGTGATGAACCACT
TGCAGGAAACGGCTCTCAATCATGTTAAGAGTGCGGGGCTTCTTAATGAAGGCTTTGGTCGTACTCCTGA
GATGTATAAAAGGGACCTTATTTGGGTTGTCGCGAAAATGCAGGTCATGGTTAACCGCTATCCTACTTGG
GGTGACACGGTTGAAGTGAATACTTGGGTTGCCAAGTCAGGGAAAAATGGTATGCGTCGTGATTGGCTCA
TAAGTGATTGCAATACAGGAGAAATTCTTACAAGGGCATCAAGTGTGTGGGTCATGATGAATCAAAAGAC
AAGAAAATTGTCAAAGATTCCAGATGAGGTTCGGCATGAGATAGAGCCTCATTTTGTGGACTCTGCTCCC
GTCATTGAAGACGATGACTGGAAACTTCCCAAGCTGGATGAGAAAACTGCTGACTCCATCCGCAAGGGTC
TAACTCCGAAGTGGAATGACTTGGATGTCAATCAGCACGTCAACAACGTGAAGTACATTGGGTGGATTCT
TGAGAGTACTCCACCAGAAGTTCTGGAGACCCAGGAGTTATGTTCCCTTACCCTGGAATACAGGCGGGAA
TGCGGAAGGGAGAGTGTGCTGGAGTCCCTCACTGCTGTGGACCCCTCTGGAAAGGGCTTTGGGCCCCAGT
TTCAGCACCTTCTGAGGCTTGAGGATGGAGGTGAGATCGTAAAGGGGAGAACTGAGTGGCGACCCAAGAC
TGCAGGTATCAATGCCACGATTGCATCTGGGGAGACCTCACCTGGAAACTCTTAG
SEQ ID NO: 69
Cuphea heterophylla (Cht) FATB3a coding DNA sequence codon optimized
for Prototheca moriformis
ATGGTGGCCACCGCCGCCTCCTCCGCCTTCTTCCCCGTGCCCTCCCCCGACACCTCCTCCCGCCCCGGCA
AGCTGGGCAACGGCTCCTCCTCCCTGCGCCCCCTGAAGCCCAAGTTCGTGGCCAACGCCGGCCTGCAGGT
GAAGGCCAACGCCTCCGCCCCCCCCAAGATCAACGGCTCCTCCGTGTCCCTGAAGTCCTGCTCCCTGAAG
ACCCACGAGGACACCCCCTCCGCCCCCCCCCCCCGCACCTTCATCAACCAGCTGCCCGACTGGTCCATGC
TGCTGGCCGCCATCACCACCGTGTTCCTGGCCGCCGAGAAGCAGTGGATGATGCTGGACTGGAAGCCCAA
GCGCCCCGACATGCTGGTGGACCCCTTCGGCCTGGGCCGCATCGTGCAGGACGGCCTGGTGTTCCGCCAG
AACTTCTCCATCCGCTCCTACGAGATCGGCGCCGACCGCACCGCCTCCATCGAGACCGTGATGAACCACC
TGCAGGAGACCGCCCTGAACCACGTGAAGTCCGCCGGCCTGCTGAACGAGGGCTTCGGCCGCACCCCCGA
GATGTACAAGCGCGACCTGATCTGGGTGGTGGCCAAGATGCAGGTGATGGTGAACCGCTACCCCACCTGG
GGCGACACCGTGGAGGTGAACACCTGGGTGGCCAAGTCCGGCAAGAACGGCATGCGCCGCGACTGGCTGA
TCTCCGACTGCAACACCGGCGAGATCCTGACCCGCGCCTCCTCCGTGTGGGTGATGATGAACCAGAAGAC
CCGCAAGCTGTCCAAGATCCCCGACGAGGTGCGCCACGAGATCGAGCCCCACTTCGTGGACTCCGCCCCC
GTGATCGAGGACGACGACTGGAAGCTGCCCAAGCTGGACGAGAAGACCGCCGACTCCATCCGCAAGGGCC
TGACCCCCAAGTGGAACGACCTGGACGTGAACCAGCACGTGAACAACGTGAAGTACATCGGCTGGATCCT
GGAGTCCACCCCCCCCGAGGTGCTGGAGACCCAGGAGCTGTGCTCCCTGACCCTGGAGTACCGCCGCGAG
TGCGGCCGCGAGTCCGTGCTGGAGTCCCTGACCGCCGTGGACCCCTCCGGCAAGGGCTTCGGCCCCCAGT
TCCAGCACCTGCTGCGCCTGGAGGACGGCGGCGAGATCGTGAAGGGCCGCACCGAGTGGCGCCCCAAGAC
CGCCGGCATCAACGGCACCATCGCCTCCGGCGAGACCTCCCCCGGCAACTCCTGA
SEQ ID NO: 70
Cuphea heterophylla (Cht) FATB3b (C67G, H72Q, L128F, N1791 variant)
amino acid sequence
MVATAASSAFFPVPSPDTSSRPGKLGNGSSSLRPLKPKFVANAGLQVKANASAPPKINGSSVSLKSGSLK
TQEDTPSAPPPRTFINQLPDWSMLLAAITTVFLAAEKQWMMLDWKPKRPDMLVDPFGFGRIVQDGLVFRQ
NFSIRSYEIGADRTASIETVMNHLQETALNHVKSAGLLIEGFGRTPEMYKRDLIWVVAKMQVMVNRYPTW
GDTVEVNTWVAKSGKNGMRRDWLISDCNTGEILTRASSVWVMMNQKTRKLSKIPDEVRHEIEPHFVDSAP
VIEDDDWKLPKLDEKTADSIRKGLTPKWNDLDVNQHVNNVKYIGWILESTPPEVLETQELCSLTLEYRRE
CGRESVLESLTAVDPSGKGFGPQFQHLLRLEDGGEIVKGRTEWRPKTAGINGTIASGETSPGNS*
SEQ ID NO: 71
Cuphea heterophylla (Cht) FATB3b (C67G, H72Q, L128F, N179I variant)
coding DNA sequence
ATGGTGGCCACCGCTGCAAGTTCTGCATTCTTCCCGGTGCCATCCCCGGACACCTCCTCTAGACCGGGAA
AGCTCGGAAATGGGTCATCAAGCTTGAGGCCCCTCAAGCCCAAATTTGTTGCCAATGCTGGGCTGCAGGT
TAAGGCAAACGCCAGTGCCCCTCCTAAGATCAATGGTTCCTCGGTCAGTCTAAAGTCTGGCAGTCTCAAG
ACTCAGGAAGACACTCCTTCGGCTCCTCCTCCGCGGACTTTTATCAACCAGTTGCCTGATTGGAGCATGC
TTCTTGCTGCAATCACTACTGTCTTCTTGGCAGCAGAGAAGCAGTGGATGATGCTTGATTGGAAACCAAA
GAGGCCTGACATGCTTGTGGACCCGTTCGGATTTGGAAGGATTGTTCAGGATGGGCTTGTGTTCAGGCAG
AATTTTTCGATTAGGTCCTATGAAATAGGCGCTGATCGCACTGCATCTATAGAGACGGTGATGAACCACT
TGCAGGAAACGGCTCTCAATCATGTTAAGAGTGCGGGGCTTCTTATTGAAGGCTTTGGTCGTACTCCTGA
GATGTATAAAAGGGACCTTATTTGGGTTGTCGCGAAAATGCAGGTCATGGTTAACCGCTATCCTACTTGG
GGTGACACGGTTGAAGTGAATACTTGGGTTGCCAAGTCAGGGAAAAATGGTATGCGTCGTGATTGGCTCA
TAAGTGATTGCAATACAGGAGAAATTCTTACTAGAGCATCAAGTGTGTGGGTCATGATGAATCAAAAGAC
AAGAAAATTGTCAAAGATTCCAGATGAGGTTCGGCATGAGATAGAGCCTCATTTTGTGGACTCTGCTCCC
GTCATTGAAGACGATGACTGGAAACTTCCCAAGCTGGATGAGAAAACTGCTGACTCCATCCGCAAGGGTC
TAACTCCGAAGTGGAATGACTTGGATGTCAATCAGCACGTCAACAACGTGAAGTACATTGGGTGGATTCT
TGAGAGTACTCCACCAGAAGTTCTGGAGACCCAGGAGTTATGTTCCCTTACCCTGGAATACAGGCGGGAA
TGCGGAAGGGAGAGTGTGCTGGAGTCCCTCACTGCTGTGGACCCCTCTGGAAAGGGCTTTGGGCCCCAGT
TTCAGCACCTTCTGAGGCTTGAGGATGGAGGTGAGATCGTAAAGGGGAGAACTGAGTGGCGACCCAAGAC
TGCAGGTATCAATGGGACGATTGCATCTGGGGAGACCTCACCTGGAAACTCTTAG
SEQ ID NO: 72
Cuphea heterophylla (Cht) FATB3b (C67G, H72Q, L128F, N179I variant)
coding DNA sequence codon optimized for Prototheca moriformis
ATGGTGGCCACCGCCGCCTCCTCCGCCTTCTTCCCCGTGCCCTCCCCCGACACCTCCTCCCGCCCCGGCA
AGCTGGGCAACGGCTCCTCCTCCCTGCGCCCCCTGAAGCCCAAGTTCGTGGCCAACGCCGGCCTGCAGGT
GAAGGCCAACGCCTCCGCCCCCCCCAAGATCAACGGCTCCTCCGTGTCCCTGAAGTCCGGCTCCCTGAAG
ACCCAGGAGGACACCCCCTCCGCCCCCCCCCCCCGCACCTTCATCAACCAGCTGCCCGACTGGTCCATGC
TGCTGGCCGCCATCACCACCGTGTTCCTGGCCGCCGAGAAGCAGTGGATGATGCTGGACTGGAAGCCCAA
GCGCCCCGACATGCTGGTGGACCCCTTCGGCTTCGGCCGCATCGTGCAGGACGGCCTGGTGTTCCGCCAG
AACTTCTCCATCCGCTCCTACGAGATCGGCGCCGACCGCACCGCCTCCATCGAGACCGTGATGAACCACC
TGCAGGAGACCGCCCTGAACCACGTGAAGTCCGCCGGCCTGCTGATCGAGGGCTTCGGCCGCACCCCCGA
GATGTACAAGCGCGACCTGATCTGGGTGGTGGCCAAGATGCAGGTGATGGTGAACCGCTACCCCACCTGG
GGCGACACCGTGGAGGTGAACACCTGGGTGGCCAAGTCCGGCAAGAACGGCATGCGCCGCGACTGGCTGA
TCTCCGACTGCAACACCGGCGAGATCCTGACCCGCGCCTCCTCCGTGTGGGTGATGATGAACCAGAAGAC
CCGCAAGCTGTCCAAGATCCCCGACGAGGTGCGCCACGAGATCGAGCCCCACTTCGTGGACTCCGCCCCC
GTGATCGAGGACGACGACTGGAAGCTGCCCAAGCTGGACGAGAAGACCGCCGACTCCATCCGCAAGGGCC
TGACCCCCAAGTGGAACGACCTGGACGTGAACCAGCACGTGAACAACGTGAAGTACATCGGCTGGATCCT
GGAGTCCACCCCCCCCGAGGTGCTGGAGACCCAGGAGCTGTGCTCCCTGACCCTGGAGTACCGCCGCGAG
TGCGGCCGCGAGTCCGTGCTGGAGTCCCTGACCGCCGTGGACCCCTCCGGCAAGGGCTTCGGCCCCCAGT
TCCAGCACCTGCTGCGCCTGGAGGACGGCGGCGAGATCGTGAAGGGCCGCACCGAGTGGCGCCCCAAGAC
CGCCGGCATCAACGGCACCATCGCCTCCGGCGAGACCTCCCCCGGCAACTCCTGA
SEQ ID NO: 73
Cuphea viscosissima (Cvis) FATB1 amino acid sequence
MVAAAATSAFFPVPAPGTSPKPGKSGNWPSSLSPTFKPKSIPNGGFQVKANASAHPKANGSAVNLKSGSL
NTQEDTSSSPPPRAFLNQLPDWSMLLTAITTVFVAAEKQWTMLDRKSKRPDMLVDSVGLKSIVRDGLVSR
HSFSIRSYEIGADRTASIETLMNHLQETTINHCKSLGLHNDGFGRTPGMCKNDLIWVLTKMQIMVNRYPT
WGDTVEINTWFSQSGKIGMASDWLISDCNTGEILIRATSVWAMMNQKTRRFSRLPYEVRQELTPHFVDSP
HVIEDNDQKLRKFDVKTGDSIRKGLTPRWNDLDVNQHVSNVKYIGWILESMPIEVLETQELCSLTVEYRR
ECGMDSVLESVTAVDPSENGGRSQYKHLLRLEDGTDIVKSRTEWRPKNAGTNGAISTSTAKTSNGNSVS
SEQ ID NO: 74
Cuphea viscosissima (Cvis) FATB1 coding DNA sequence codon optimized
for Prototheca moriformis
ATGGTGG6CGCCGCCGCCACCTCCGCCTTCTTCCCCGTGCCCGCCCCCGGCACCTCCCCCAAGCCCGGCA
AGTCCGGCAACTGGCCCTCCTCCCTGTCCCCCACCTTCAAGCCCAAGTCCATCCCCAACGGCGGCTTCCA
GGTGAAGGCCAACGCCTCCGCCCACCCCAAGGCCAACGGCTCCGCCGTGAACCTGAAGTCCGGCTCCCTG
AACACCCAGGAGGACACCTCCTCCTCCCCCCCCCCCCGCGCCTTCCTGAACCAGCTGCCCGACTGGTCCA
TGCTGCTGACCGCCATCACCACCGTGTTCGTGGCCGCCGAGAAGCAGTGGACCATGCTGGACCGCAAGTC
CAAGCGCCCCGACATGCTGGTGGACTCCGTGGGCCTGAAGTCCATCGTGCGCGACGGCCTGGTGTCCCGC
CACTCCTTCTCCATCCGCTCCTACGAGATCGGCGCCGACCGCACCGCCTCCATCGAGACCCTGATGAACC
ACCTGCAGGAGACCACCATCAACCACTGCAAGTCCCTGGGCCTGCACAACGACGGCTTCGGCCGCACCCC
CGGCATGTGCAAGAACGACCTGATCTGGGTGCTGACCAAGATGCAGATCATGGTGAACCGCTACCCCACC
TGGGGCGACACCGTGGAGATCAACACCTGGTTCTCCCAGTCCGGCAAGATCGGCATGGCCTCCGACTGGC
TGATCTCCGACTGCAACACCGGCGAGATCCTGATCCGCGCCACCTCCGTGTGGGCCATGATGAACCAGAA
GACCCGCCGCTTCTCCCGCCTGCCCTACGAGGTGCGCCAGGAGCTGACCCCCCACTTCGTGGACTCCCCC
CACGTGATCGAGGACAACGACCAGAAGCTGCGCAAGTTCGACGTGAAGACCGGCGACTCCATCCGCAAGG
GCCTGACCCCCCGCTGGAACGACCTGGACGTGAACCAGCACGTGTCCAACGTGAAGTACATCGGCTGGAT
CCTGGAGTCCATGCCCATCGAGGTGCTGGAGACCCAGGAGCTGTGCTCCCTGACCGTGGAGTACCGCCGC
GAGTGCGGCATGGACTCCGTGCTGGAGTCCGTGACCGCCGTGGACCCCTCCGAGAACGGCGGCCGCTCCC
AGTACAAGCACCTGCTGCGCCTGGAGGACGGCACCGACATCGTGAAGTCCCGCACCGAGTGGCGCCCCAA
GAACGCCGGCACCAACGGCGCCATCTCCACCTCCACCGCCAAGACCTCCAACGGCAACTCCGTGTCCTGA
SEQ ID NO: 75
Cuphea viscosissima (Cvis) FATB2 amino acid sequence
MVATAASSAFFPVPSADTSSRPGKLGNGPSSFSPLKPKSIPNGGLQVKASASAPPKINGSSVGLKSGGLK
THDDAPSAPPPRTFINQLPDWSMLLAAITTAFLAAEKQWMMLDRKPKRLDMLEDPFGLGRVVQDGLVFRQ
NFSIRSYEIGADRTASIETVMNHLQETALNHVKTAGLSNDGFGRTPEMYKRDLIWVVAKMQVMVNRYPTW
GDTVEVNTWVAKSGKNGMRRDWLISDCNTGEILTRASSVWVMMNQKTRKLSKIPDEVRREIEPHFVDSAP
VIEDDDRKLPKLDEKSADSIRKGLTPRWNDLDVNQHVNNAKYIGWILESTPPEVLETQELCSLTLEYRRE
CGRESVLESLTAVDPSGEGYGSQFQHLLRLEDGGEIVKGRTEWRPKNAGINGVVPSEESSPGDYS
SEQ ID NO: 76
Cuphea viscosissima (Cvis) FATB2 coding DNA sequence codon optimized
for Prototheca moriformis
ATGGTGGCCACCGCCGCCTCCTCCGCCTTCTTCCCCGTGCCCTCCGCCGACACCTCCTCCCGCCCCGGCA
AGCTGGGCAACGGCCCCTCCTCCTTCTCCCCCCTGAAGCCCAAGTCCATCCCCAACGGCGGCCTGCAGGT
GAAGGCCTCCGCCTCCGCCCCCCCCAAGATCAACGGCTCCTCCGTGGGCCTGAAGTCCGGCGGCCTGAAG
ACCCACGACGACGCCCCCTCCGCCCCCCCCCCCCGCACCTTCATCAACCAGCTGCCCGACTGGTCCATGC
TGCTGGCCGCCATCACCACCGCCTTCCTGGCCGCCGAGAAGCAGTGGATGATGCTGGACCGCAAGCCCAA
GCGCCTGGACATGCTGGAGGACCCCTTCGGCCTGGGCCGCGTGGTGCAGGACGGCCTGGTGTTCCGCCAG
AACTTCTCCATCCGCTCCTACGAGATCGGCGCCGACCGCACCGCCTCCATCGAGACCGTGATGAACCACC
TGCAGGAGACCGCCCTGAACCACGTAAGACCGCCGGCCTGTCCAACGACGGCTTCGGCCGCACCCCCGA
GATGTACAAGCGCGACCTGATCTGGGTGGTGGCCAAGATGCAGGTGATGGTGAACCGCTACCCCACCTGG
GGCGACACCGTGGAGGTGAACACCTGGGTGGCCAAGTCCGGCAAGAACGGCATGCGCCGCGACTGGCTGA
TCTCCGACTGCAACACCGGCGAGATCCTGACCCGCGCCTCCTCCGTGTGGGTGATGATGAACCAGAAGAC
CCGCAAGCTGTCCAAGATCCCCGACGAGGTGCGCCGCGAGATCGAGCCCCACTTCGTGGACTCCGCCCCC
GTGATCGAGGACGACGACCGCAAGCTGCCCAAGCTGGACGAGAAGTCCGCCGACTCCATCCGCAAGGGCC
TGACCCCCCGCTGGAACGACCTGGACGTGAACCAGCACGTGAACAACGCCAAGTACATCGGCTGGATCCT
GGAGTCCACCCCCCCCGAGGTGCTGGAGACCCAGGAGCTGTGCTCCCTGACCCTGGAGTACCGCCGCGAG
TGCGGCCGCGAGTCCGTGCTGGAGTCCCTGACCGCCGTGGACCCCTCCGGCGAGGGCTACGGCTCCCAGT
TCCAGCACCTGCTGCGCCTGGAGGACGGCGGCGAGATCGTGAAGGGCCGCACCGAGTGGCGCCCCAAGAA
CGCCGGCATCAACGGCGTGGTGCCCTCCGAGGAGTCCTCCCCCGGCGACTACTCCTGA
SEQ ID NO: 77
Cuphea viscosissima (Cvis) FATB3 amino acid sequence
MVAAAASSAFFSFPTPGTSPKPGKFGNWPSSLSIPFNPKSNHNGGIQVKANASAHPKANGSAVSLKAGSL
ETQEDTSSPSPPPRTFISQLPDWSMLVSAITTVFVAAEKQWTMLDRKSKRPDVLVEPFVQDGVSFRQSFS
IRSYEIGVDRTASIETLMNIFQETSLNHCKSLGLLNDGFGRTPEMCKRDLIWVVTKMQIEVNRYPTWGDT
IEVTTWVSESGKNGMSRDWLISDCHSGEILIRATSVWAMMNQKTRRLSKIPDEVRQEIVPYFVDSAPVIE
DDRKLHKLDVKTGDSIRNGLTPRWNDFDVNQHVNNVKYIAWLLKSVPTEVFETQELCGLTLEYRRECRRD
SVLESVTAMDPSKEGDRSLYQHLLRLENGADIALGRTEWRPKNAGATGAVSTGKTSNGNSVS
SEQ ID NO: 78
Cuphea viscosissima (Cvis) FATB3 coding DNA sequence codon optimized
for Prototheca moriformis
ATGGTGGCCGCCGCCGCCTCCTCCGCCTTCTTCTCCTTCCCCACCCCCGGCACCTCCCCCAAGCCCGGCA
AGTTCGGCAACTGGCCCTCCTCCCTGTCCATCCCCTTCAACCCCAAGTCCAACCACAACGGCGGCATCCA
GGTGAAGGCCAACGCCTCCGCCCACCCCAAGGCCAACGGCTCCGCCGTGTCCCTGAAGGCCGGCTCCCTG
GAGACCCAGGAGGACACCTCCTCCCCCTCCCCCCCCCCCCGCACCTTCATCTCCCAGCTGCCCGACTGGT
CCATGCTGGTGTCCGCCATCACCACCGTGTTCGTGGCCGCCGAGAAGCAGTGGACCATGCTGGACCGCAA
GTCCAAGCGCCCCGACGTGCTGGTGGAGCCCTTCGTGCAGGACGGCGTGTCCTTCCGCCAGTCCTTCTCC
ATCCGCTCCTACGAGATCGGCGTGGACCGCACCGCCTCCATCGAGACCCTGATGAACATCTTCCAGGAGA
CCTCCCTGAACCACTGCAAGTCCCTGGGCCTGCTGAACGACGGCTTCGGCCGCACCCCCGAGATGTGCAA
GCGCGACCTGATCTGGGTGGTGACCAAGATGCAGATCGAGGTGAACCGCTACCCCACCTGGGGCGACACC
ATCGAGGTGACCACCTGGGTGTCCGAGTCCGGCAAGAACGGCATGTCCCGCGACTGGCTGATCTCCGACT
GCCACTCCGGCGAGATCCTGATCCGCGCCACCTCCGTGTGGGCCATGATGAACCAGAAGACCCGCCGCCT
GTCCAAGATCCCCGACGAGGTGCGCCAGGAGATCGTGCCCTACTTCGTGGACTCCGCCCCCGTGATCGAG
GACGACCGCAAGCTGCACAAGCTGGACGTGAAGACCGGCGACTCCATCCGCAACGGCCTGACCCCCCGCT
GGAACGACTTCGACGTGAACCAGCACGTGAACAACGTGAAGTACATCGCCTGGCTGCTGAAGTCCGTGCC
CACCGAGGTGTTCGAGACCCAGGAGCTGTGCGGCCTGACCCTGGAGTACCGCCGCGAGTGCCGCCGCGAC
TCCGTGCTGGAGTCCGTGACCGCCATGGACCCCTCCAAGGAGGGCGACCGCTCCCTGTACCAGCACCTGC
TGCGCCTGGAGAACGGCGCCGACATCGCCCTGGGCCGCACCGAGTGGCGCCCCAAGAACGCCGGCGCCAC
CGGCGCCGTGTCCACCGGCAAGACCTCCAACGGCAACTCCGTGTCCTGA
