MICROGININ PRODUCING PROTEINS AND NUCLEIC ACIDS ENCODING A MICROGININ GENE CLUSTER AS WELL AS METHODS FOR CREATING NOVEL MICROGININS

Abstract

The invention provides for nucleic acid molecules enabling the synthesis of microginin and microginin analogues. The invention also provides for methods for identifying microginins as well creating microginins which may not be found in nature.

Description

TECHNICAL FIELD

The present invention relates to the fields of chemistry, biology, biochemistry, molecular biology. The invention provides for novel nucleic acid molecules enabling the synthesis of microginin and microginin analogues. Microginin finds an application in therapeutics. The invention thus extends into the field of mammalian therapeutics and drug development.

INTRODUCTION

Cyanobacteria and Microginin

Cyanbacteria are gram-negative bacteria. Due to their ability to perform photosynthesis they were long thought to belong to the plant kingdom and were formerly classified as blue-green algae. Cyanbacteria have adapted to almost all ecological niches. Most of strains known up to date are found in fresh water lakes and oceans. In the last few years cyanobacteria have been recognised as a source for biologically active natural compounds.

Cyanobacteria are a group of microscopic organisms somewhere “in between” algae and bacteria and they are found in freshwater and marine areas throughout the world. Scientifically, they are considered to be bacteria, but because they can perform photosynthesis, they also used to be classified as “blue-green algae”.

Cyanobacterial peptides (cyanopeptides) are among the most ubiquitously found potentially hazardous natural products in surface waters used by humans. Though these substances are natural in origin, eutrophication (i.e. excessive loading with fertilising nutrients) has caused massive cyanobacterial proliferation throughout Europe. Thus, cyanopeptides now occur with unnatural frequency and concentration.

A large group among the diverse cyanopeptides are the oligopeptides (peptides with a molecular weight of <2KD). But while specific cyanopeptides—e.g. microcystins and nodularins—are well studied and recognised as being causative for many animal poisonings and human illness, a substantial and increasing body of evidence points toward a decisive role of other potentially toxic cyanopeptides in the causation of both acute and chronic human illnesses.

Freshwater and marine cyanobacteria are known to produce a variety of bioactive compounds, among them potent hepatotoxins and neurotoxins. Many of the toxic species of cyanobacteria tend to massive proliferation in eutrophicated water bodies and thus have been the cause for considerable hazards for animal and human health. One of the most widespread bloom-forming cyanobacteria is the genus Microcystis, a well-known producer of the hepatotoxic peptide microcystin. Microcystins are a group of closely related cyclic heptapeptides sharing the common structure. So far, more than 80 derivatives of microcystins have been identified, varying largely by the degree of methylation, peptide sequence, and toxicity.

The traditional botanical code describes the genus Microcystis as a coccal, unicellular cyanobacterium that grows as mucilaginous colonies of irregularly arranged cells (under natural conditions, while strain cultures usually grow as single cells). According to this tradition, morphological criteria such as size of the individual cells, colony morphology, and mucilage characteristics are used for species delimitation within Microcystis (i.e., morphospecies). Microcystin-producing strains as well as strains that do not synthesize microcystin have been reported for all species within the genus Microcystis. However, whereas most field samples and strains of Microcystis aeruginosa and Microcystis viridis studied to date were found to contain microcystins, strains of M. wesenbergii, M. novaceckii, and M. ichthyoblabe have only sporadically been reported to contain microcystins.

Beside microcystins, various other linear and cyclic oligopeptides such as anabaenopeptins, aeruginosins, microginins and cyanopeptolins are found within the genus Microcystis (Namikoshi, M., and K. L. Rinehart. 1996. Bioactive compounds produced by cyanobacteria. J. Ind. Microbiol. 17:373-384.).

Similar to microcystins, these peptides possess unusual amino acids like 3-amino-6-hydroxy-2-piperidone (Ahp) in cyanopeptolins, 2-carboxy-6-hydroxyoctahydroindol (Choi) in aeruginosin-type molecules or 3-amino-2 hydroxy-decanoic acid (Ahda) in microginins and numerous structural variants also exist within these groups. These peptides show diverse bioactivities, frequently protease inhibition (Namikoshi, M., and K. L. Rinehart. 1996. Bioactive compounds produced by cyanobacteria. J. Ind. Microbiol. 17:373-384).

The occurrence of both microcystins and other oligopeptides such as anabaenopeptins, microginins and cyanopeptolins in natural Microcystis populations was recently demonstrated. It is well known that the species and genotype composition in natural Microcystis populations is heterogeneous, and both microcystin- and non-microcystin-containing strains have been isolated from the same sample. Just as strains producing microginin and strains not producing microginin have been found. These results suggest a considerable diversity of genotypes with different oligopeptide patterns in natural Microcystis populations.

By typing single Microcystis colonies, it was possible in 1999 to show for the first time that the actual peptide diversity in a natural population of this genus is extremely high. Many of the substances detected belong to well-known groups of cyanobacterial peptides like microcystins, anabaenopeptins, microginins, cyanopeptolins, and aeruginosins, of which many have been discovered in Microcystis spp. In addition, numerous unknown components have been detected in such colonies. However, the origin of these unknown components has yet to be investigated, since besides the observed epiphytic cyanobacteria and algae, heterotrophic bacteria are also known to be present in Microcystis colonies. Chemical screening of cyanobacterial samples (both from field samples and from culture strains) has demonstrated a wide variety of substances: e.g. an almost monospecific bloom of Planktothrix agardhii contained as many as 255 different substances, most of which were oligopeptides.

Thus, it may be concluded, that the situation with respect to the assignment of the capability of microginin production to certain species and strains, i.e. also a true understanding of the genotypes and species involved as well as their evolution has to date, not been possible. In fact PEPCY a research project supported by the European Commission concluded that present information shows that one species or “morphotype” (i.e. individuals with the same morphological characteristics) may comprise a range of genotypes that encode for different “chemotypes” (i.e. morphologically indistinguishable individuals containing different cyanopeptides).

Ace Inhibitors and Microginin

ACE catalyses the conversion of angiotensin I into angiotensin II within the mammalian renin-angiotensin system, leading to arterial stenosis, which in turn causes an increase of blood pressure. ACE inhibitors counteract this process and therefore play a role in human medicine as blood pressure lowering agents. Microginin is an important drug candidate for ACE inhibition. So far only 30 structural variants of microginin are known, making clinical development difficult.

Microginins are characterized by a decanoic acid derivate, 3-amino-2-hydroxy-decanoic acid (Ahda) at the N-terminus and a predominance of two tyrosine units at the C-terminus. They vary in length from 4 to 6 amino acids with the variability occurring at the C-terminal end (Microginins, zinc metalloprotease inhibitors from the cyanobacterium Microcystis aeruginosa, 2000, Tetrahedron 56:8643-8656). In the past it has only been possible by means of synthesis of 3-amino-2-hydroxy-decanoic acid to chemically generate microginin variants (J Org. Chem. 1999 Apr. 16; 64(8):2852-2859. Acylnitrene Route to Vicinal Amino Alcohols. Application to the Synthesis of (−)-Bestatin and Analogues. Bergmeier S C, Stanchina D M.) Alternatively cyanobacterial strains were screened for microginin activity, which was tedious and time consuming. It has so far not been possible to screen for strains efficiently due to the lack of species understanding and a methodology of efficiently distinguishing microginin producers from non-producers (see above). Further it was not possible to easily and efficiently alter and thus develop microginins in order to provide for a variety of lead compounds from which better ACE-inhibitors may be developed.

BRIEF DESCRIPTION OF THE INVENTION

From Microcystis aeruginosa a cluster of genes, spanning about 30 kbps has been isolated encoding a hybrid synthetase composed of non-ribosomal peptide synthetases (NRPS), polyketide synthases (PKS) and tailoring enzyme which as the inventors show is responsible for the biosynthesis of microginin. The strain from which this nucleic acid was first isolated by G. C. Kürzinger from Lake Pehlitz 1977].

The inventors provide for a biological system enabling not only the production of micoginins, the heterologous expression of microginin, but also a system for modifying microginin and thus developing so far unknown variants of microginin. The invention further provides for nucleic acids and methods for identifying strains which have the ability to produce microginin.

In particular the invention relates to one or more nucleic acids encoding a microginin synthetase enzyme complex with the following activities: an adenylation domain (A*) wherein, the adenylation domain comprises a peptide sequence according to SEQ ID NO. 1, an acyl carrier protein (ACP), an elongation module (EM) of polyketide synthases (PKS) comprising the following activities: (i) ketoacylsynthase (KS), (ii) acyl transferase (AT) (iii) acyl carrier protein (ACP2), an aminotransferase (AMT), three to five elongation modules (EM) of non-ribosomal peptide synthetases (NRPS) comprising the following activities: (i) condensation domain (C), (ii) adenylation domain (A), (iii) thiolation domain (T) and a thioesterase (TE).

DETAILED DESCRIPTION OF THE INVENTION

As outlined above the invention in particular relates to one or more nucleic acids encoding a microginin synthetase enzyme complex with the following activities: an adenylation domain (A*) wherein, the adenylation domain comprises a peptide sequence according to SEQ ID NO. 1, an acyl carrier protein (ACP), an elongation module (EM) of polyketide synthases (PKS) comprising the following activities: (i) ketoacylsynthase (KS), (ii) acyl transferase (AT) (iii) acyl carrier protein (ACP 2), an aminotransferase (AMT), three to five elongation modules (EM) of non-ribosomal peptide synthetases (NRPS) comprising the following activities: (i) condensation domain (C), (ii) adenylation domain (A), (iii) thiolation domain (T) and a thioesterase (TE).

The inventors have found that microginin is the product of non-ribosomal synthesis. It is important to understand that microginin as previously identified in nature may also in part have been the product of ribosomal synthesis and further processed via various enzymatic reactions.

It is important to note that the nucleic acid claimed herein, i.e. a microginin synthetase enzyme complex may also be present in organisms other organisms than Microcystis sp., such as Nostoc, Anabaena, Plankthotrix or Oscillatoria. The term microginin shall thus not limit the invention to such nucleic acids producing synthetase enzyme complexes resulting in peptides officially termed “microginin”.

Herein, an adenylation domain (A*) is understood to activate octanoic acid as an acyl adenylate and an acyl carrier protein (ACP) is understood to bind the octanoic acid adenylate as a thioester.

An elongation module (EM) of polyketide synthases (PKS) is also known e.g. from the Jamaicamide synthetase gene cluster isolated from Lyngbya majuscula (Chem. Biol. Vol. 11, 2004 pp 817-833. Structure and Biosynthesis of the Jamaicamides, new mixed polyketide-peptide neurotoxin from the marine cyanobacterium Lyngbya majuscula) herein comprises at least the following activities: (i) ketoacylsynthase (KS), (ii) acyl transferase (AT) and (iii) acyl carrier protein (ACP2). The AT is responsible for the recognition of malonyl-CoA, the KS is responsible for the Claisen-type-condensation of the activated octanoic acid adenylate with malonyl-CoA and the ACP2 is responsible for binding of the resulting decanoic acid.

An aminotransferase (AMT) performs the β-amination of the decanoic acid.

The nucleic acid according to the invention may have three to five elongation modules (EM) of non-ribosomal peptide synthetases (NRPS) comprising at least the following activities: (i) condensation domain (C), (ii) adenylation domain (A), (iii) thiolation domain (T). The A is responsible for the activation of carboxyl groups of amino acids, the T is responsible for the binding and the transport of the activated intermediate, the C is responsible for the condensation of the activated amino acids with the growing peptide chain.

Finally the nucleic acid according to the invention shall contain a thioesterase (TE) activity which performs the cleavage of the final product from the synthetase complex.

One may envision that the nucleic acid according to the invention is present in a vector or a bacterial chromosome, in which case one may envision that the portions designated above while being in one cell need not all, be in, or on, one molecule. It is essential to the invention however, that a cell meant to produce microginin synthetase enzyme complex contains the activities designated above in order to produce an enzyme complex according to the invention which in turn may produce a microginin. Thus, the invention also encompasses derivatives of the nucleic acid molecule as outlined above having the function of a microginin synthetase enzyme complex.

The molecule is characterized by a special adenylation domain (A*) which is unusual in that it is not similar to known adenlyation domains found in other molecules encoding non-ribosomal enzyme complexes such as the microcystin synthetase gene cluster (Chem. Biol. Vol. 7 2000, pp 753-764: Structural organisation of microcystin synthesis in Microcystis aeruginosa PCC 7806: In integrated peptide-polyketide-synthetase system) Molecules encompassed herein are those which carry this adenylation domain (A*) as depicted in SEQ ID NO. 1 and at least an ACP whereby this ACP may stem from another known non-ribosomal enzyme complex, at least one EM of PKS whereby this EM may stem from another known non-ribosomal enzyme complex comprising at least the following activities: (i) KS, (ii) AT (iii) ACP, an AMT whereby this AMT may stem from another known non-ribosomal enzyme complex three to five EMs comprising at least the following activities: (i) C, (ii) A, (iii) T whereby these EMs may stem from another known non-ribosomal enzyme complex and a TE whereby this TE may stem from another known non-ribosomal enzyme complex. Chimeras whereby parts of the above are on one or more vectors and or integrated in chromosomes are equally encompassed by the invention as long as all the components are in one cell.

The invention also pertains to isolated nucleic acid molecules encoding a microginin synthetase enzyme complex comprising an adenylation domain which is 85% identical to SEQ ID NO. 1, more preferred 90% identical to SEQ ID NO. 1 most preferred 95% identical to SEQ ID NO. 1. Sequence identity herein is in percent of total sequence of the adenylation domains when aligned with conventional nucleotide alignment software, such as the best fit and or pileup programs of the GCG package

The invention also pertains to a microginin synthetase enzyme protein complex with the following activities: an adenylation domain (A*) wherein, the adenylation domain comprises a peptide sequence according to SEQ ID NO. 1, an acyl carrier protein (ACP), an elongation module (EM) of polyketide synthases (PKS) comprising the following activities: (i) ketoacylsynthase (KS), (ii) acyl transferase (AT) (iii) acyl carrier protein (ACP 2), an aminotransferase (AMT), three to five elongation modules (EM) of non-ribosomal peptide synthetases (NRPS) comprising the following activities: (i) condensation domain (C), (ii) adenylation domain (A), (iii) thiolation domain (T) and a thioesterase (TE).

The invention in particular also relates to a nucleic acid molecule encoding an adenylation domain (A*) wherein, the adenylation domain comprises a peptide sequence according to SEQ ID NO. 1.

The invention in particular also relates to a peptide molecule, an adenylation domain (A*) wherein, the molecule comprises a peptide sequence according to SEQ ID NO. 1.

The invention in particular also relates to a nucleic acid molecule encoding an adenylation domain (A*) wherein, the molecule comprises a nucleic acid sequence according to SEQ ID NO. 25.

In a preferred embodiment of the invention the nucleic acid additionally and optionally comprises sequences encoding the following activities or domains: a monooxygenase (MO), an integrated N-methyltransferase domain (MT) within one or more elongation modules (EM) of NRPS, a non-integrated N-methyltrasferase (MT), a modifying activity (MA) wherein, said MA is selected from the group comprising the following activities: halogenase, sulfatase, glycosylase, racemase, O-methyltransferase and C-methyltransferase, two or more peptide repeat spacer sequences (SP) consisting of one or more repeats of being either glycine rich or proline and leucine rich, located adjacently upstream and downstream of the MO and/or another MA.

Herein MO is an enzyme catalyzing the hydroxylation of the decanoic acid, an integrated N-methyltransferase domain (MT) within one or more elongation modules (EM) of NRPS catalyses the methylation of the amide bond by the respective module and a non-integrated N-methyltrasferase (MT) catalyzes the methylation of an amino group of the microginin.

The term modifying enzyme stands for numerous enzymes such enzymes may add groups or create bonds, in a preferred embodiment MA is selected from the group comprising the following activities: halogenase, sulfatase, glycosylase, racemase, O-methyltransferase and C-methyltransferase.

Nucleic acids encoding two or more peptide repeat spacer sequences (SP) consisting of one or more repeats being either glycine rich or proline and leucine rich have astonishingly been found by the inventors to aid in integration of novel MAs into existing microginin synthetase enzyme complexes. By means of placing such SPs adjacently to MAs the inventors are able to create microginin synthetase enzyme complexes (MSEC) comprising activities previously not found in MSECs. This in turn allows for the creation of novel microginins with potentially novel therapeutic properties. Thus the invention relates to nucleic acids encoding two or more peptide repeat spacer sequences (SP) consisting of one or more repeats being either glycine rich or proline and leucine rich may be positioned adjacently to a MA such as but not limited to a halogenase, a sulfatase, a glycosylase, a racemase, an O-methyltransferase or a C-methyltransferase. These SPs aid in ensuring that the “foreign” activity “works” in the enzyme complex. The inventors have found, that this is due to the lack of secondary structures in the SP peptide chains.

The nucleic acid according to the invention in a preferred embodiment optionally comprises the following sequences, nucleic acid sequences encoding protein sequences as follows:

An adenylation domain (A*) according to SEQ ID NO. 1, an acyl carrier protein (ACP) according to SEQ ID NO. 2, an elongation module of polyketide synthases responsible for the activation and the condensation of malonyl-Co A: (i) ketoacylsynthase domain (KS) according to SEQ ID NO. 3, (ii) acyl transferase domain (AT) according to SEQ ID NO. 4, an acyl carrier protein domain (ACP 2) according to SEQ ID NO. 5, an aminotransferase (AMT) according to SEQ ID NO. 6, an elongation modules of non-ribosomal peptide synthetases responsible for the activation and condensation of alanin: (i) condensation domain (C) according to SEQ ID NO. 7, (ii) adenylation domain (A) according to SEQ ID NO. 8, (iii) thiolation domains (T) according to SEQ ID NO. 9, an elongation modules of non-ribosomal peptide synthetases responsible for the activation and condensation of leucin: (i) condensation domain (C 2) according to SEQ ID NO. 10, (ii) adenylation domain (A 2) according to SEQ ID NO. 11, (iii) thiolation domain (T 2) according to SEQ ID NO. 12, an elongation modules of non-ribosomal peptide synthetases responsible for the activation and condensation of tyrosine 1: (i) condensation domain (C 3) according to SEQ ID NO. 13, (ii) adenylation domain (A 3) according to SEQ ID NO. 14 (iii) thiolation domain (T 3) according to SEQ ID NO. 15, an elongation modules of non-ribosomal peptide synthetases responsible for the activation and condensation of tyrosine 2: (i) condensation domain (C 4) according to SEQ ID NO. 16, (ii) adenylation domain (A 4) according to SEQ ID NO. 17, (iii) thiolation domain (T 4) according to SEQ ID NO. 18, a thioesterase (TE) according to SEQ ID NO. 19, a monooxygenase (MO) according to SEQ ID NO. 20, two or more peptide repeat spacer sequences (SP1/SP2) according to SEQ ID NO. 21 and 22, an integrated N-methyltransferase domain (MT) within the elongation module (EM) of the NRPS responsible for the activation and condensation of leucin according to SEQ ID 23 and a non-integrated N-methyltrasferase (MT 2) according to SEQ ID NO. 24.

As outlined above, the minimal requirement according to the invention is a nucleic acid encoding a microginin synthetase enzyme complex with the following activities: an adenylation domain (A*) wherein, the adenylation domain comprises a peptide sequence according to SEQ ID NO. 1, an ACP according to SEQ ID NO. 2, an elongation module (EM) of polyketide synthases (PKS) comprising the following activities: (i) ketoacylsynthase (KS) according to SEQ ID NO. 3, (ii) acyl transferase (AT) according to SEQ ID NO 4, (iii) acyl carrier protein (ACP 2) according to SEQ ID NO. 5, an aminotransferase (AMT) according to SEQ ID NO. 6, three to five elongation modules (EM) of non-ribosomal peptide synthetases (NRPS) comprising the following activities: (i) condensation domain (C) according to SEQ ID NO. 7, (ii) adenylation domain (A) according to SEQ ID NO. 8, (iii) thiolation domain (T) according to SEQ ID NO. 9 and a thioesterase (TE) according to SEQ ID NO. 10. A molecule comprising the above sequences is preferred herein.

The invention explicitly also relates to analogs hereto, additionally comprising, e.g. other activities and/or spacer regions both transcribed and non-transcribed.

It is apparent to those skilled in the art, that amino acids may be exchanged maintaining the enzymatic activity required. Thus, the invention also relates to molecules with sequences which are not identical to those outlined above however, altered only in so far as the enzymatic activity desired is retained.

The nucleic acid according to the invention may contain nucleic acids selected from the group comprising: an adenylation domain (A*) according to SEQ ID NO. 25, an acyl carrier protein (ACP) according to SEQ ID NO. 26, an elongation module of polyketide synthases encoding for the activation and the condensation of malonyl-Co A: (i) ketoacylsynthase domain (KS) according to SEQ ID NO. 27, (ii) acyl transferase domain (AT) according to SEQ ID NO. 28, (iii) acyl carrier protein domain (ACP 2) according to SEQ ID NO. 29, an aminotransferase (AMT) according to SEQ ID NO. 30, an elongation modules of non-ribosomal peptide synthetases encoding for the activation and condensation of alanin: (i) condensation domain (c) according to SEQ ID NO. 31, (ii) adenylation domain (A) according to SEQ ID NO. 32, (iii) thiolation domain (T) according to SEQ ID NO. 33, an elongation modules of non-ribosomal peptide synthetases encoding for the activation and condensation of leucin: (i) condensation domain (C 2) according to SEQ ID NO. 34, (ii) adenylation domain (A 2) according to SEQ ID NO. 35, (iii) thiolation domain (T 2) according to SEQ ID NO. 36, elongation modules of non-ribosomal peptide synthetases encoding for the activation and condensation of tyrosine 1: (i) condensation domains (C 3) according to SEQ ID NO. 37, (ii) adenylation domains (A 3) according to SEQ ID NO. 38, (iii) thiolation domains (T 3) according to SEQ ID NO. 39, elongation modules of non-ribosomal peptide synthetases encoding for the activation and condensation of tyrosine 2: (i) condensation domains (C 4) according to SEQ ID NO. 40, (ii) adenylation domains (A 4) according to SEQ ID NO. 41, (iii) thiolation domains (T 4) according to SEQ ID NO. 42, a thioesterase (TE) according to SEQ ID NO. 43, a monooxygenase (MO) according to SEQ ID NO. 44, two or more peptide repeat spacer sequences (SP1/2) according to SEQ ID NO. 45 and 46, an integrated N-methyltransferase domain (MT) within the elongation module (EM) of the NRPS encoding for the activation and condensation of leucin according to SEQ ID 47 and a non-integrated N-methyltrasferase (MT 2) according to SEQ ID NO. 48.

As outlined above, the minimal requirement according to the invention is a nucleic acid encoding a microginin synthetase enzyme complex with the following activities: an adenylation domain (A*) wherein, the adenylation domain is a nucleic acid sequence according to SEQ ID NO. 25, an ACP with a nucleic acid sequence according to SEQ ID NO. 26, an elongation module (EM) of polyketide synthases (PKS) comprising the following activities: (i) ketoacylsynthase (KS) with a nucleic acid sequence according to SEQ ID NO. 27, (ii) acyl transferase (AT) with a nucleic acid sequence according to SEQ ID NO 28, (iii) acyl carrier protein (ACP 2) with a nucleic acid sequence according to SEQ ID NO. 29, an aminotransferase (AMT) with a nucleic acid sequence according to SEQ ID NO. 30, three to five elongation modules (EM) of non-ribosomal peptide synthetases (NRPS) comprising the following activities: (i) condensation domain (C) with a nucleic acid sequence according to SEQ ID NO. 31, (ii) adenylation domain (A) with a nucleic acid sequence according to SEQ ID NO. 32, (iii) thiolation domain (T) with a nucleic acid sequence according to SEQ ID NO. 33 and a thioesterase (TE) with a nucleic acid sequence according to SEQ ID NO. 43. A molecule comprising the above sequences is preferred herein.

The invention also relates to nucleic acid molecules with sequences which are not identical to those outlined above however, altered only in so far as the enzymatic activity desired is retained. I particular one skilled in the art will know that positions in nucleic acid triplets may “wobble” and these positions may thus be altered with no influence on the peptide sequence. Further multiple amino acids are encoded by more than one DNA triplet. One skilled in the art will know that one may alter such triplets maintaining the amino acid sequence. Thus said sequences are equally encompassed by the invention.

The invention also pertains to isolated nucleic acid molecules encoding a microginin synthetase enzyme complex comprising an adenylation domain which is 85% identical to SEQ ID NO. 25, more preferred 90% identical to SEQ ID NO. 1 most preferred 95% identical to SEQ ID NO. 1. Sequence identity herein is in percent of total sequence of the adenylation domains when aligned with a conventional amino acid alignment software such as the best fit and or pileup programs of the GCG package.

In a preferred embodiment the one or more nucleic acids according to the invention are organized in sequence parts encoding the microginin synthetase enzyme complex in an upstream to downstream manner as depicted in FIG. 1. In a particularly preferred embodiment the activities and domains are arranged as shown and on one molecule.

The nucleic acid molecule may be part of a vector. Such vectors are in particular, bacterial artificial chromosomes (BAC), Cosmids or Fosmids, and Lambda vectors, Preferred plasmid vectors which are able to replicate autonomously in cyanobacteria are derived from the pVZ vectors. Preferred fosmid vectors which are able to replicate autonomously in cyanobacteria are derived from the pCC1FOS™ and pCC2FOS™ vectors (Epicentre Biotechnologies). The integration of the nucleic acid according to the invention into the vector is a procedure known to those skilled in the art (Molecular Cloning: A Laboratory manual, 1989, Cold Spring Harbour Laboratory Press) or in the manuals of manufactures of kits for creation of genomic libraries (e.g. Epicenter Biotechnologies).

In a preferred embodiment the invention concerns a microorganism transformed with a nucleic acid according to the invention. The nucleic acid according to the invention may integrated into the chromosome of the host organism or may present on a separate vector (see also examples). It is preferred that the phototrophic cyanobacterial host organism is selected for the group comprising: Synechocystis sp., Synechococcus sp., Anabaena sp., Nostoc sp., Spirulina sp., Microcystis sp. . . . Cells are cultured as follows:

Media: Bg 11 (for cultivation of cyanobacteria)
Aeration: air containing 0.3-3.0% carbon dioxide
Light intensity: 40-100 μE/m²*s (diameter of illuminated culture vessels of photobioreactor d=4-12 cm)
Cell density at harvest: OD_750nm 1-2
And if the host is Microcystis aeruginosa:
Light quality: Additional red light illumination with 25 μE/m²*s for 24-48 hours before harvesting.

It is preferred that the heterotrophic host organism is selected for the group comprising: E. coli and Bacillus sp. due to a more suitable GC content and codon usage than other heterotrophic bacteria.

In case of using E. coli for the heterologues expression of the microginin synthetase a phosphopanthetein transferase (Ppt) has to be co-expressed in order to enable the synthesis of microginin. The co-expression of the Ppt from a microginin producing strain would be preferred. Other Ppt's with a broad specificity even from heterotophic organisms like Bacillus sp. are also suitable.

In one embodiment of the invention the invention relates to a method of producing a microginin, comprising culturing a cell under conditions under which the cell will produce microginin, wherein said cell comprises a nucleic acid encoding a recombinant microginin, according to the invention, and wherein said cell does not produce the microginin in the absence of said nucleic acid.

The inventors have identified nucleic acid sequences which for the first time make it possible to detect nucleic acids encoding a microginin synthetase enzyme complex. This has been extremely difficult, due to the fact that other gene clusters which encode non-ribosomal protein producing complexes share sequence similarity with the present cluster claimed herein. Such primers or probes according to the invention are selected from the group of, a) nucleic acid according to SEQ ID NO. 49 (Primer A), b) nucleic acid according to SEQ ID NO. 50 (Primer B), c) nucleic acid according to SEQ ID NO. 51 (Primer C), d) nucleic acid according to SEQ ID NO. 52 (Primer D), e) nucleic acid according to SEQ ID NO. 53 (Primer E), f) nucleic acid according to SEQ ID NO. 54 (Primer F), g) nucleic acid according to SEQ ID NO. 55 (Primer G), h) nucleic acid according to SEQ ID NO. 56 (Primer H), i) nucleic acid according to SEQ ID NO. 57 (Primer I) and j) nucleic acid according to SEQ ID NO. 58 (Primer J). It is known to one skilled in the art that such primers or probes may be altered slightly and still accomplishes the task of specifically detecting the desired target sequence. Such alterations in sequence are equally encompassed by the invention. The primers or probes according to the invention may be applied in hybridization reactions and/or amplification reactions. Such reactions are known to one skilled in the art.

The invention also concerns a method for detecting a microginin synthetase gene cluster in a sample wherein, one or more of the nucleic acids according to the invention are, applied in an amplification and/or a hybridization reaction.

In a preferred embodiment of the method according to the invention primers D and F or H and J or E and I or E and A are added to a PCR reaction mixture comprising a sample and wherein, presence of an amplification product represents presence of microginin synthetase gene cluster and absence of an amplification product represents absence of a microginin synthetase gene cluster. As can be seen from the examples (example 3 below), certain combinations are preferred. Samples may be isolated DNA, prokaryotic cells stemming from plates or liquid cultures.

When performing an amplification reaction with primers D and F the most preferred amplification conditions are as follows: a) denaturing, b) 48° C. annealing and c) elongation (product size: 675 bp). These temperatures may vary a bit in the range of 2-8 degrees C.

When performing an amplification reaction with primers H and J the most preferred amplification conditions are as follows: a) denaturing, b) 54° C. annealing and c) elongation (product size: 1174 bp). These temperatures may vary a bit in the range of 2-8 degrees C.

When performing an amplification reaction with primers E and I the most preferred amplification conditions are as follows: a) denaturing, b) 56° C. annealing and c) elongation (product size: 1279 bp). These temperatures may vary a bit in the range of 2-8 degrees C.

When performing an amplification reaction with primers E and A the most preferred amplification conditions are as follows: a) denaturing, b) 57° C. annealing and c) elongation (product size: 621 bp). These temperatures may vary a bit in the range of 2-8 degrees C. Molarity is most commonly 0.2-1.0 μM for the primers. Buffers and other reagents depending on polymerase used.

When performing hybridisation reactions the above nucleic acids are usually labeled. Such labels may be radioactive or non-radioactive, such as fluorescent. The nucleic acid primers or probes may be applied, e.g. for the screening of libraries.

The invention also relates to antibodies against a peptide according to SEQ ID NO. 1 (A*). The creation of such antibodies is known to one skilled in the art. The antibodies may be polyclonal or monoclonal. Such antibodies may be labeled or non-labeled, they may also be altered in other form, such as humanized.

The inventors have astonishingly found that newly identified peptide repeat spacer sequences (SP) may be placed adjacently to MAs I in order to create novel hybrid gene clusters. These SPs act by spacing the novel activity or domain so that it is functionally active in the microginin synthetase enzyme complex.

The invention thus, further relates to nucleic acids encoding a peptide repeat spacer sequence (SP) wherein, the peptide sequence comprises at least 4 glycin amino acids per single repeat unit (SRU) or, at least 5 proline and/or leucin amino acids per SRU, A SRU within the SP is between 7 and 15 amino acids in length and, the SP comprises between 2 and 10 SRUs.

The invention further relates to peptides of a peptide repeat spacer sequence (SP) wherein, the peptide sequence comprises at least 4 glycin amino acids or, at least 5 proline and/or leucin amino acids, the single repeat unit (SRU) within the SP is between 7 and 15 amino acids in length and, the SP comprises between 2 and 10 SRU. In a preferred embodiment of the invention the SRU is between 9 and 13 amino acids in length in a particularly preferred embodiment the SRU is eleven amino acids in length. In a preferred embodiment the SP comprises between 3 and 9 SRU.

In a preferred embodiment the nucleic acid encoding the peptide repeat spacer sequence (SP) according to the invention, encodes a peptide SRU as shown in SEQ ID NO. 20 or SEQ ID NO. 21. In a further embodiment the peptide repeat spacer sequence (SP) according to the invention, comprises or contains a sequence as shown in SEQ ID NO. 20 or SEQ ID NO. 21. In a further embodiment the nucleic acid according to the invention has a sequence as laid down in SEQ ID NO. 43 or SEQ ID NO. 44.

Not only by means of the above mentioned SPs but in particular because of these the inventors are able to create enzyme complexes resulting in microginin variants which may not be found in nature. This is an essential aspect of the present invention. The invention provides for, for the first time a simple method of producing recombinant microginin variants comprising, modifying the nucleic acid according to the invention in vitro or in vivo, growing a recombinant cell comprising said recombinantly modified nucleic acid encoding a microginin synthetase under conditions which lead to synthesis of a microginin and, recovering the synthesized microginin.

In a preferred embodiment of said method according to the invention, said modifying of said nucleic acid may be an action selected from the group of one or more of the following actions: a) inactivation of one or more of the MTs present, b) substitution of one or more of the MTs present with a halogenase, a sulfatase, a glycosylase, a racemase, an O-methyltransferase or a C-methyltransferase, c) inactivation of the MO, d) substitution of the MO with a halogenase, a sulfatase, a glycosylase, a racemase, an O-methyltransferase or a C-methyltransferase, e) inactivation of the AMT, f) substitution of the AMT with a halogenase, a sulfatase, a glycosylase, a racemase, an O-methyltransferase or a C-methyltransferase, g) inactivation of the PKS module, h) substitution of the entire PKS module with an alternative PKS module and/or substitution of one or more of the domains (KS, AT, ACP) therein, i) inactivation of the A* domain, j) substitution of the A* domain with alternative A domains, k) inactivation of one or more of the NRPS modules and 1) substitution of one or more of the NRPS modules with alternative NRPS modules and/or substitution of one or more of the domains (C, A, T) therein.

Halogenases, sulfatases, glycosylases, racemases, O-methyltransferases or C-methyltransferases are known from prokaryotes. These enzymes are encoded by genes of the secondary metabolism in particular NRPS/PKS systems.

Alternative PKS-systems, entire modules as well as single domains (KS, AT, ACP) are found in cyanobacteria as well as Actinomycetes, Myxobacteria, Bacillus among the bacteria.

Alternative NRPS-systems, entire modules as well as single domains (C, A, T) are found in cyanobacteria as well as Actinomycetes, Myxobacteria, Bacillus among the bacteria.

In a preferred embodiment the above are from cyanobacteria.

It is important to note, that said inactivation and/or substitution may done in many ways, e.g. inactivation may imply deleting the complete activity or domain, or may imply inactivation by means of a single nucleotide exchange.

The methods are known to those skilled in the art and comprise basic molecular biological methods such as DNA isolation, restriction digestion, ligation, transformation, amplification etc.

In a preferred embodiment said alternative modules or domains which are used for substitution of the original module or domain, additionally may comprise one or more SP nucleic acids according to the invention located adjacently upstream of the module or domain used for substitution and one or more SP nucleic acids according the invention located adjacently downstream of the module or domain used for substitution. Thus, in this embodiment of the invention a construct is made comprising the domain which is to be entered into the original nucleic acid according to the invention, further comprising one or more SPs located adjacently in an upstream and downstream manner. This construct is then ligated into the original microginin synthetase encoding nucleic acid. The resultant construct is then brought into a host by means of transformation for either a) integration into the host chromosome or b) with a self-replicating vector.

The polypeptides, i.e. proteins can be any of those described above but with not more than 10 (e.g., not more than: 10, nine, eight, seven, six, five, four, three, two, or one) conservative substitutions. Conservative substitutions are known in the art and typically include substitution of, e.g. one polar amino acid with another polar amino acid and one acidic amino acid with another acidic amino acid. Accordingly, conservative substitutions preferably include substitutions within the following groups of amino acids: glycine, alanine, valine, proline, isoleucine, and leucine (non polar, aliphatic side chain); aspartic acid and glutamic acid (negatively charged side chain); asparagine, glutamine, methionine, cysteine, serine and threonine (polar uncharged side chain); lysine, histidine and arginine; and phenylalanine, tryptophane and tyrosine (aromatic side chain); and lysine, arginine an histidine (positively charged side chain). It is well known in the art how to determine the effect of a given substitution, e.g. on pK₁ etc. All that is required of a polypeptide having one or more conservative substitutions is that it has at least 50% (e.g., at least: 55%; 60%; 65%, 70%; 75%; 80%; 85%; 90%; 95%; 98%; 99%; 99.5%; or 100% or more) of the ability of the unaltered protein according to the invention.

In preferred embodiments the polynucleotides, i.e. nucleic acids of the present invention also comprise nucleic acid molecules which are at least 85%, preferably 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to those claimed herein.

The determination of percent identity between two sequences is accomplished using the mathematical algorithm of Karlin and Altschul (1993) Proc. Natl. Acad. Sci. USA 90: 5873-5877. Such an algorithm is incorporated into the BLASTN and BLASTP programs of Altschul et al. (1990) J. Mol. Biol. 215: 403-410. BLAST nucleotide searches are performed with the BLASTN program, score=100, word length=12, to obtain nucleotide sequences homologous to the nucleic acids according to the invention. BLAST protein searches are performed with the BLASTP program, score=50, wordlength=3, to obtain amino acid sequences homologous to the EPO variant polypeptide, respectively. To obtain gapped alignments for comparative purposes, Gapped BLAST is utilized as described in Altschul et al. (1997) Nucleic Acids Res. 25: 3389-3402. When utilizing BLAST and Gapped BLAST programs, the default parameters of the respective programs are used.

FIGURES

FIG. 1 depicts the structure of microginin.

FIG. 2 depicts the microginin synthetase gene cluster and the biosynthetic pathway of microginin.

EXAMPLES

Example 1

Method for Detecting Gene Clusters According to the Invention

Strains carrying a gene cluster encoding a microginin synthetase complex can be distinguished from strains not carrying such a gene cluster performing a PCR reaction using RedTaq ReadyMix PCR Reaction Mix with MgCl₂ (Sigma) and primer pairs and the corresponding annealing temperatures as described in Claims 11-12. In particular the PCR conditions are as follows: an initial denaturation for 1 minutes at 95° C., followed by 30 cycles of denaturation at 95° C. for 30 seconds, elongation at said annealing temperatures for 30 seconds and extension at 72° C. for 1 kb of product size.

Example 2

Method for Optimised Cultivation of Microginin Producing Microcystis spp.

Strains. Media: Bg 11 (for cultivation of cyanobacteria)
Aeration: air containing 0.3-3.0% carbon dioxide
Light intensity: 40-100 μE/m²*s (diameter of illuminated culture vessels of photobioreactor d=4-12 cm)
Light quality: Additional red light illumination with 25 μE/m²*s for 24-48 hours before harvesting.
Cell density at harvest: OD_750nm 1-2

Tables

TABLE 1
SEQ ID      MTINYGDLQEPFNKFSTLVELLRYRASSQPERLAYIFLRDGEIEEARLTYGELDQKARAI
NO. 1 A*    AAYLQSLEAEGERGLLLYPPGLDFISAFFGCLYAGVVAIPAYPPRRNQNLLRLQAIIADS
            QARFTFTNAALFPSLKNQWAKDPELGAMEWIVTDEIDHHLREDWLEPTLEKNSLAFLQYT
            SGSTGTPKGVMVSHHNLLINSADLDRGWGHDQDSVMVTWLPTFHDMGLIYGVIQPLYKGF
            LCYMMSPASFMERPLRWLQALSDKKATHSAAPNFAYDLCVRKIPPEKRATLDLSHWCMAL
            NGAEPVRAEVLKKFAEAFQVSGFKATALCPGYGLAEATLKVTAVSYDSPPYFYPVQANAL
            EKNKIVGATETDTNVQTLVGCGWTTIDTQIVIVNPETLKPCSPEIVGEIWVSGSTIAQGY
            WGKPQETQETFQAYLADTGAGPFLRTGDLGFIKDGELFITGRLKEIILIRGRNNYPQDIE
            LTVQNSHPALRPSCGAAFTVENKGEEKLVVVQEVERTWLRKVDIDEVKRAIRKAVVQEYD
            LQVYAIALIRTGSLPKTSSGKIQRRSCRAKFLEGSLEILG
SEQ ID      MSTEIPNDKKQPTLTKIQNWLVAYMTEMMEVDEDEIDLSVPFDEYGLDSSMAVALIADLE
NO. 2       DWLRRDLHRTLIYDYPTLEKLAKQVSEP
ACP
SEQ ID      MEPIAIIGLACRFPGADNPEAFWQLMRNGVDAIADIPPERWDIERFYDPTPATAKKMYSR
NO. 3       QGGFLKNVDQFDPQFFRISPLEATYLDPQQRLLLEVTWEALENAAIVPETLAGSQSGVFI
KS          GISDVDYHRLAYQSPTNLTAYVGTGNSTSIAANRLSYLFDLRGPSLAVDTACSSSLVAVH
            LACQSLQSQESNLCLVGGVNLILSPETTVVFSQARMIAPDSRCKTFDARADGYVRSEGCG
            VVVLKRLRDAIQDGDRILAVIEGSAVNQDGLSNGLTAPNGPAQQAVIRQALANAQVKPAQ
            ISYVEAHGTGTELGDPIEVKSLKAVLGEKRSLDQTCWLGSVKTNIGHLEAAAGMAGLIKV
            VLCLQHQEIPPNLHFQTLNPYISLADTAFAIPTQAQPWRTKPPKSGENGVERRLAGLSSF
            GFGGTNSHVIL
SEQ ID      VFLFAGQGSQYVGMGRQLYETQPIFRQTLDRCAEILRPHLDQPLLEILYPADPEAETASF
NO. 4 AT    YLEQTAYTQPTLFAFEYALAQLWRSWGIEPAAVIGHSVGEYVAATVAGALSLEEGLTLIA
            KRAKLMQSLPKNGTMIAVFAAEERVKAVIEPYRTDVAIAAVNGPENFVISGKAPIIAEII
            IHLTAAGIEVRPLKVSHAFHSHLLEPILDSLEQEAAAISYQPLQIPLVANLTGEVLPEGA
            TIEARYWRNHARNPVQFYGSIQTLIEQKFSLFLEVSPKPTLSRLGQQCCPERSTTWLFSL
            APPQEEEQSLLNSLAILYDSQGAE
SEQ ID      ITLQTLVGNLLQLSPADVNVHTPFLEMGADSIVMVEAVRRIENTYNVKIAMRQLFEELST
NO. 5       LDALATYL
ACP 2
SEQ ID      KEMLYPIVAQRSQGSRIWDVDGNEYIDMTMGQGVTLFGHQPDFIMSALQSQLTEGIHLNP
NO. 6       RSPIVGEVAALICELTGAERACFCNSGTEAVMAAIRIARATTGRSKIALFEGSYHGHADG
AMT         TLFRNQIIDNQLHSFPLALGVPPSLSSDVVVLDYGSAEALNYLQTQGQDLAAVLVEPIQS
            GNPLLQPQQFLQSLRQITSQMGIALIFDEMITGFRSHPGGAQALFGVQADIATYGKVVAG
            GMPIGVIAGKAHYLDSIDGGMWRYGDKSYPGVDRTFFGGTFNQHPLAMVAARAVLTHLKE
            QGPGLQQQLTERTAALADTLNHYFQAEEVPIKIEQFSSFFRFALSGNLDLLFYHMVEKGI
            YVWEWRKHFLSTAHTEADLAQFVQAVKDSITELR
SEQ ID      GGDQVPLTEAQRQLWILAQLGDNGSVAYNQSVTLQLSGPLNPVAMNQAIQQISDRHEALR
NO. 7 C     TKINAQGDSQEILPQVEINCPILDFSLDQASAQQQAEQWLKEESEKPFDLSQGSLVRWHL
            LKLEPELHLLVLTAHHIISDGWSMGVILRELGELYSAKCQGVTANLKTPKQFRELIEWQS
            QPSQGEELKKQQAYWLATLADPPVLNLPTDKPRPALPSYQANRRSLTLDSQFTEKLKQFS
            RKQGCTLLMTLLSVYNILVHRLTGQDDILVGLPASGRGLLDSEGMVGYCTHFLPIRSQLA
SEQ ID      TYSELNCRANQLAHYLQKLGVGPEVLVGILVERSLEMIVGLLGILKAGGAYVPLDPDYPP
NO. 8 A     ERLQFMLEDSQFFLLLTQQHLLESFAQSSETATPKIICLDSDYQIISQAKNINPENSVTT
            SNLAYVIYTSGSTGKPKGVMNNHVAISNKLLWVQDTYPLTTEDCILQKTPFSFDVSVWEL
            FWPLLNGARLVFAKPNGHKDASYLVNLIQEQQVTTLHFVSSMLQLFLTEKDVEKCNSLKR
            VICSGEALSLELQERFFARLVCELHNLYGPTEAAIHVTFWQCQSDSNLKTVPIGRPIANI
            QIYILDSHLQPVPIGVIGELHIGGVGLARGYLNRPELTAEKFIANPFASLDPPLTPLDKG
            GDESYKTFKKGGEQPSRLYKTGDLARYLPDGKIEYLGRIDNQVKIRGFRIELGEIEAVLL
            SHPQVREAVV
SEQ ID      EAIAAIFGQVLKLEKVGIYDNFFEIGGNSLQATQVISRLRESFALELPLRRLFEQPTVAD
NO. 9 T     LALAV
SEQ ID      PRDGQLPLSFAQSRLWFLYQLEGATGTYNMTGALSLSGPLQVEALKQALRTIIQRHEPLR
NO. 10 C 2  TSFQSVDGVPVQVINPYPVWELAMVDLTGKETEAEKLAYQESQTPFDLTNSPLLRVTLLK
            LQPEKHILLINMHHIISDGWSIGVFVRELSHLYRAFVAGKEPTLPILPIQYADFAVWQRE
            WLQGKVLAAQLEYWKRQLADAPPLLELPTDRPRPAIQTFQGKTERFELDRKLTQELKALS
            QQSGCTLFMTLLAAFGVVLSRYSGQTDIVIGSAIANRNRQDIEGLIGFFVNTLALRLDLS
SEQ ID      TYGELNHRANQLAHYLQSLGVTKEQIVGVYLERSLEMAIGFLGILKAGAAYLPIDPEYPS
NO. 11 A 2  VRTQFILEDTQLSLLLTQAELAEKLPQTQNKIICLDRDWPEITSQPQTNLDLKIEPNNLA
            YCIYTSGSTGQPKGVLISHQALLNLIFWHQQAFEIGPLHKATQVAGIAFDATVWELWPYL
            TTGACINLVPQNILLSPTDLRDWLLNREITMSFVPTPLAEKLLSLDWPNHSCLKTLLLGG
            DKLHFYPAASLPFQVINNYGPTENTVVATSGLVKSSSSHHFGTPTIGRPIANVQIYLLDQ
            NLQPVPIGVPGELHLGGAGLAQGYLNRPELTAEKFIANPFDPPLTPLDKGGEEPSKLYKT
            GDLARYLPDGNVEFLGRIDNQVKIRGFRIETGEIEAVLSQYFLLAESVV
SEQ ID      AQLTQIWSEVLGLERIGVKDNFFELGGHSLLATQVLSRINSAFGLDLSVQIMFESPTIAG
NO. 12 T 2  IAGYI
SEQ ID      ARDGHLPLSFAQQRLWFLHYLSPDSRSYNTLEILQIDGNLNLTVLEQSLGELINRHEIFR
NO. 13 C 3  TTFPTVSGEPIQKIALPSRFQLKVDNYQDLDENEQSAKIQQVAELEAGQAFDLTVGPLIQ
            FKLLQLSPQKSVLLLKMHHIIYDGWSFGILIRELSALYEAFLKNLANPLPALSIQYADFA
            VWQRQYLSGEVLDKQLNYWQEQLATVSPVLTLPTDRPRPAIQTFQGGVERFQLDQNVTQG
            LKKLGQDQVATLFMTLLAGFGVLLSRYSGQSDLMVGSPIANRNQAAIEPLIGFFANTLAL
            RINLS
SEQ ID      TYTELNHRANQLAHYLQTLGVGAEVLVGISLERSLEMIIGLLGILKVGGAYLPLDPDYPT
NO. 14 A 3  ERLQLMLEDSQVPFLITHSSLLAKLPPSQATLICLDHIQEQISQYSPDNLQCQLTPANLA
            NVIYTSGSTGKPKGVMVEHKGLVNLALAQIQSFAVNHNSRVLQFASFSFDACISEILMTF
            GSGATLYLAQKDALLPGQPLIERLVKNGITHVTLPPSALVVLPQEPLRNLETLIVAGEAC
            SLDLVKQWSIDRNFFNAYGPTEASVCATIGQCYQDDLKVTIGKAIANVQIYILDAFLQPV
            PVGVSGELYIGGVGVARGYLNRPELTQEKFIANPFSNDPDSRLYKTGDLARYLPDGNIEY
            LGRIDNQVKIRGFRIELGEIEAVLSQCPDVQNTAV
SEQ ID      EILAQIWGQVLKIERVSREDNFFELGGHSLLATQVMSRLRETFQVELPLRSLFTAPTIAE
NO. 15 T 3  LALTI
SEQ ID      NDSANLPLSFAQQRLWFLDQLEPNSAFYHVGGAVRLEGTLNITALEQSLKEIINRHEALR
NO. 16 C 4  TNFITIDGQATQIIHPTINWRLSVVDCQNLTDTQSLEIAEAEKPFNLAQDCLFRATLFVR
            SPLEYHLLVTMHHIVSDGWSIGVFFQELTHLYAVYNQGLPSSLTPIKIQYADFAVWQRNW
            LQGEILSNQLNYWREQLANAPAFLPLPTDRPRPAIQTFIGSHQEFKLSQPLSQKLNQLSQ
            KHGVTLFMTLLAAFATLLYRYTGQADILVGSPIANRNRKEIEGLIGFFVNTLVLRLSLD
SEQ ID      TYAELNHQANQLVHYLQTLGIGPEVLVAISVERSLEMIIGLLAILKACGAYLPLAPDYPT
NO. 17 A 4  ERLQFMLEDSQASFLITHSSLLEKLPSSQATLICLDHIQEQISQYSPDNLQSELTPSNLA
            NVIYTSGSTGKPKGVMVEHRGLVNLASSQIQSFAVKNNSRVLQFASFSFDACISEILMTF
            GSGATLYLAQKNDLLPGQPLMERLEKNKITHVTLPPSALAVLPKKPLPNLQTLIVAGEAC
            PLDLVKQWSVGRNFFNAYGPTETSVCATIGQCYQDDLKVTIGKAIANVQIYILDAFLQPV
            PIGVPGELYIGGVGVARGYLNRPELTAERFIPNPFDPPLTPLKKGGDKSYETFKKGEEQP
            SKLYKTGDLARYLPDGNIEYLGRIDNQVKIRGFRIELGEIEAVLSQCPDVQNTAV
SEQ ID      LQLAQIWSEILGINNIGIQENFFELGGHSLLAVSLINRIEQKLDKRLPLTSLFQNGTIAS
NO. 18 T 4  LAQLL
SEQ ID      TPFFAVHPIGGNVLCYADLARNLGTKQPFYGLQSLGLSELEKTVASIEEMAMIYIEAIQT
NO. 19      VQASGPYYLGGWSMGGVIAFEIAQQLLTQGQEVALLALIDSYSPSLLNSVNREKNSANSL
TE          TEEFNEDINIAYSFIRDLASIFNQEISFSGSELAHFTSDELLDKFITWSQETNLLPSDFG
            KQQVKTWFKVFQINHQALSSYSPKTYLGRSVFLGAEDSSIKNPGWHQ
SEQ ID      FSLYYFGSYEAEFNPNKYNLLFEGAKFGDRAGFTALWIPERHFHAFGGFSPNPSVLAAAL
NO. 20      ARETKQIQLRSGSVVLPLHNSIRVAEEWAVVDNLSQGRVGIAFASGWHPQDFVLAPQSFG
MO          QHRELMFQEIETVQKLWRGEAITVPDGKGQRVEVKTYPQPMQSQLPSWITIVNNPDTYIR
            AGAIGANILTNLMGQSVEDLARNIALYRQSLAEHGYDPASGTVTVLLHTFVGKDLEQVRE
            QARQPFGQYLTSSVGLLQNMVKSQGMKVDFEQLRDEDRDFLLASAYKRYTETSALIGTPE
            SCRQIIDHLQSIGVDEVACFIDFGVDEQTVLANLPYLQSLKDLYQ
SEQ ID      IDPPLTPLDKGIDPPLTPLDKGIDPPLTPLDKG
NO. 21
SP 1
SEQ ID      PYQGGLGGDQSPYQGGLGGDQSPYQGGLGGDQSPYQGGLGGDQSPYQGGLGGDQSPYQGE
NO. 22      LGGDQSPYQGGLGGDQV
SP 2
SEQ ID      PASEMREWVENTVSRILAFQPERGLEIGCGTGLLLSRVAKHCLEYWATDYSQGAIQYVER
NO. 23      VCNAVEGLEQVKLRCQMADNFEGIALHQFDTVVLNSIIQYFPSVDYLLQVLEGAINVIGE
MT          RGQIFVGDVRSLPLLEPYHAAVQLAQASDSKTVEQWQQQVRQSVAGEEELVIDPTLFLAL
            KQHFPQISWVEIQPKRGVAHNELTQFRYDVTLHLETINNQALLSGNPTVITWLNWQLDQL
            SLTQIKDKLLTDKPELWGIRGIPNQRVEEALKIWEWVENAPDVETVEQLKKLLKQQVDTG
            INPEQVWQLAESLGYTAHLSWWESSQDGSFDVIFQRNSEAEDSKKLTLSKLAFWDEKPFK
            IKPWSDYTNNPLRGKLVQKLIP
SEQ ID      MTNYGKSMSHYYDLVVGHKGYNKDYATEVEFIHNLVETYTTEAKSILYLGCGTGYHAALL
NO. 24      AQKGYSVHGVDLSAEMLEQAKTRIEDETIASNLSFSQGNICEIRLNRQFNVVLALFHVVN
MT 2        YQTTNQNLLATFATVKNHLKAGGIFICDVSYGSYVLGEFKSRPTASILRLEDNSNGNEVT
            YISELNFLTHENIVEVTHNLWVTNQENQLLENSRETHLQRYLFKPEVELLADACELTVLD
            AMPWLEQRPLTNIPCPSVCFVIGHKTTHSA
SEQ ID      ATGACTATTAACTATGGTGATCTGCAAGAACCCTTTAATAAATTCTCAACCCTAGTTGAA
NO. 25      TTACTCCGTTATCGGGCAAGCAGTCAACCGGAACGCCTCGCCTATATTTTTCTGCGAGAC
A* nucl     GGAGAAATCGAAGAAGCTCGTTTAACCTATGGGGAACTGGATCAAAAGGCTAGGGCGATC
acid        GCCGCTTATCTACAATCCTTAGAAGCCGAGGGCGAAAGGGGTTTACTGCTCTATCCCCCA
            GGACTAGATTTTATTTCAGCTTTTTTTGGTTGTTTATATGCGGGAGTCGTTGCCATTCCC
            GCCTATCCACCCCGACGGAATCAAAACCTTTTGCGTTTACAGGCGATTATTGCCGATTCT
            CAAGCCCGATTTACCTTCACCAATGCCGCTCTATTTCCCAGTTTAAAAAACCAATGGGCT
            AAAGACCCTGAATTAGGAGCAATGGAATGGATTGTTACCGATGAAATTGACCATCACCTC
            AGGGAGGATTGGCTAGAACCAACCCTCGAAAAAAACAGTCTCGCTTTTCTACAATACACC
            TCTGGTTCAACGGGAACTCCAAAGGGAGTAATGGTCAGTCACCATAATTTGTTGATTAAT
            TCAGCCGATTTAGATCGTGGTTGGGGCCATGATCAAGATAGCGTAATGGTCACTTGGCTA
            CCGACCTTCCATGATATGGGTCTGATTTATGGGGTTATTCAGCCTTTGTACAAAGGATTT
            CTTTGTTACATGATGTCCCCTGCCAGCTTTATGGAACGACCGTTACGTTGGTTACAGGCC
            CTTTCTGATAAAAAAGCAACCCATAGTGCGGCCCCCAACTTTGCCTACGATCTTTGTGTG
            CGGAAAATTCCCCCTGAAAAACGGGCTACGTTAGACTTAAGCCATTGGTGCATGGCCTTA
            AATGGGGCCGAACCCGTCAGAGCGGAGGTACTTAAAAAGTTTGCGGAGGCCTTTTCAAGTT
            TCTGGTTTCAAAGCCACAGCCCTTTGTCCTGGCTACGGTTTAGCAGAAGCCACCCTGAAA
            GTTACGGCGGTTAGTTATGACAGTCCCCCTTACTTTTATCCCGTTCAGGCTAATGCTTTA
            GAAAAAAATAAGATTGTGGGAGCCACTGAAACCGATACCAATGTGCAGACCCTCGTGGGC
            TGCGGCTGGACAACGATTGATACTCAAATCGTCATTGTCAATCCTGAAACCCTGAAACCT
            TGCTCCCCTGAAATTGTCGGCGAAATTTGGGTATCAGGTTCAACAATCGCCCAAGGCTAT
            TGGGGAAAACCTCAAGAGACTCAGGAAACCTTTCAAGCTTATTTGGCAGATACAGGAGCC
            GGGCCTTTTCTGCGAACAGGAGACTTGGGCTTCATTAAAGATGGTGAATTGTTTATCACA
            GGTCGGCTCAAGGAAATTATTCTGATTCGAGGACGCAATAATTATCCCCAGGATATTGAA
            TTAACCGTCCAAAATAGTCATCCCGCTCTGCGTCCCAGTTGTGGGGCTGCTTTTACCGTT
            GAAAATAAGGGCGAAGAAAAGCTCGTGGTCGTTCAGGAAGTGGAGCGCACCTGGCTCCGT
            AAGGTAGATATAGATGAGGTAAAAAGAGCCATTCGTAAAGCTGTTGTCCAGGAATATGAT
            TTACAGGTTTATGCGATCGCGCTGATCAGGACTGGCAGTTTACCAAAAACCTCTAGCGGT
            AAAATTCAGCGTCGTAGCTGTCGGGCCAAATTTTTAGAGGGAAGCCTGGAAATTTTGGGC
            TAA
SEQ ID      ATGTCCACAGAAATCCCAAACGACAAAAAACAACCGACCCTAACGAAAATTCAAAACTGG
NO. 26      TTAGTGGCTTACATGACAGAGATGATGGAAGTGGACGAAGATGAGATTGATCTGAGCGTT
ACP nucl    CCCTTTGATGAATATGGTCTCGATTCTTCTATGGCAGTTGCTTTGATCGCTGATCTAGAG
acid        GATTGGTTACGACGAGATTTACATCGCACCCTGATCTACGATTATCCAACTCTAGAAAAG
            TTGGCTAAACAGGTTAGTGAACCCTGA
SEQ ID      ATGGAACCCATCGCAATTATTGGTCTTGCTTGCCGCTTTCCAGGGGCTGACAATCCAGAA
NO. 27      GCTTTCTGGCAACTCATGCGAAATGGGGTGGATGCGATCGCCGATATTCCTCCTGAACGT
KS nucl     TGGGATATTGAGCGTTTCTACGATCCCACACCTGCCACTGCCAAGAAGATGTATAGTCGC
acid        CAGGGCGGTTTTCTAAAAAATGTCGATCAATTTGACCCTCAATTTTTCCGAATTTCTCCC
            CTAGAAGCCACCTATCTAGATCCTCAACAAAGACTGCTACTGGAAGTCACCTGGGAAGCC
            TTAGAAAATGCTGCCATTGTGCCTGAAACCTTAGCTGGTAGCCAATCAGGGGTTTTTATT
            GGTATCAGTGATGTGGATTATCATCGTTTGGCTTATCAAAGTCCTACTAACTTGACCGCC
            TATGTGGGTACAGGCAACAGCACCAGTATTGCGGCTAACCGTTTATCATATCTGTTTGAT
            TTGCGTGGCCCCAGTTTGGCCGTAGATACCGCTTGCTCTTCTTCCCTCGTCGCCGTTCAC
            TTGGCCTGTCAGAGTTTGCAAAGTCAAGAATCGAACCTCTGCTTAGTGGGGGGAGTTAAT
            CTCATTTTGTCGCCAGAGACAACCGTTGTTTTTTCCCAAGCGAGAATGATCGCCCCCGAC
            AGTCGTTGTAAAACCTTTGACGCGAGGGCCGATGGTTATGTGCGCTCGGAAGGCTGTGGA
            GTAGTCGTACTTAAACGTCTTAGGGATGCCATTCAGGACGGCGATCGCATTTTAGCAGTG
            ATTGAAGGTTCCGCGGTGAATCAGGATGGTTTAAGTAATGGACTCACGGCCCCTAATGGC
            CCTGCTCAACAGGCGGTGATTCGTCAGGCCCTGGCAAATGCCCAGGTAAAACCGGCCCAG
            ATTAGCTATGTCGAAGCCCATGGCACGGGGACAGAATTGGGGGATCCGATCGAAGTTAAA
            TCTCTGAAAGCGGTTTTGGGTGAAAAGCGATCGCTCGATCAAACCTGTTGGCTCGGTTCT
            GTGAAAACCAACATTGGTCATTTAGAAGCGGCGGCGGGAATGGCGGGTCTGATTAAAGTC
            GTTCTCTGCCTACAACACCAAGAAATTCCCCCTAATCTCCACTTTCAAACCCTTAATCCC
            TATATTTCCCTAGCTGACACAGCTTTTGCGATTCCCACTCAGGCTCAACCCTGGCGGACC
            AAACCCCCTAAGTCTGGTGAAAACGGTGTCGAACGACGTTTAGCAGGACTCAGTTCCTTT
            GGGTTTGGGGGGACAAATTCCCATGTGATTCTC
SEQ ID      GTTTTTCTATTTGCCGGTCAAGGTTCTCAATATGTAGGTATGGGTCGTCAACTGTACGAA
NO. 28      ACCCAACCCATCTTTCGCCAAACCTTGGATCGCTGTGCTGAAATCCTGCGACCCCATTTA
AT nucl     GATCAACCCCTCTTAGAAATTCTTTATCCTGCTGACCCAGAAGCCGAAACAGCGAGTTTT
acid        TACCTAGAGCAGACTGCCTATACCCAACCCACTTTATTCGCATTCGAGTATGCCCTAGCA
            CAGTTATGGCGTTCCTGGGGAATAGAACCGGCGGCAGTAATTGGTCACAGTGTCGGTGAA
            TATGTGGCGGCCACCGTTGCCGGAGCCTTAAGTCTAGAAGAAGGATTAACGCTAATTGCC
            AAACGGGCAAAACTGATGCAGTCTCTCCCCAAGAATGGGACAATGATCGCCGTTTTTGCC
            GCAGAAGAGCGGGTTAAAGCTGTTATTGAGCCTTATAGGACTGATGTAGCGATCGCTGCT
            GTTAATGGACCAGAAAATTTTGTTATTTCAGGAAAAGCGCCGATTATTGCTGAGATTATC
            ATTCATTTAACGGCAGCAGGAATAGAAGTTCGTCCTCTCAAAGTTTCCCATGCTTTTCAC
            TCGCACCTGTTGGAGCCAATTTTAGATTCCTTAGAACAGGAAGCTGCTGCTATTTCCTAC
            CAACCCCTGCAAATTCCCTTAGTTGCTAATTTAACGGGGGAAGTTCTACCAGAAGGAGCA
            ACGATTGAGGCTCGTTACTGGCGAAATCATGCACGCAACCCTGTACAATTTTATGGGAGT
            ATCCAAACGCTGATCGAGCAGAAATTCAGTCTTTTTTTAGAAGTTAGCCCTAAACCGACT
            TTATCTCGATTGGGTCAACAATGTTGTCCAGAAAGATCGACCACTTGGCTATTTTCCCTC
            GCCCCTCCTCAAGAAGAAGAACAAAGCCTACTAAATAGTTTGGCGATTCTCTATGATTCC
            CAAGGAGCCGAA
SEQ ID      ATCACATTGCAAACCCTAGTGGGAAATTTACTGCAATTGTCCCCTGCTGATGTCAATGTT
NO. 29      CATACACCTTTCCTGGAGATGGGGGCAGATTCCATTGTCATGGTTGAGGCGGTCAGACGG
ACP 2       ATTGAGAATACCTATAACGTTAAAATTGCTATGCGTCAGTTATTTGAGGAGTTATCTACT
nucl acid   TTAGATGCTTTAGCTACTTATTTA
SEQ ID      AAAGAGATGCTTTATCCCATTGTGGCCCAACGTTCTCAAGGATCAAGAATTTGGGATGTG
NO. 30      GACGGTAATGAATATATTGATATGACGATGGGGCAAGGGGTAACGCTGTTTGGGCATCAA
AMT         CCAGACTTCATTATGTCGGCCCTACAAAGCCAACTCACTGAAGGCATTCATCTCAATCCG
nucl acid   CGATCGCCAATTGTGGGAGAAGTGGCCGCCTTAATTTGTGAACTAACAGGAGCCGAACGA
            GCTTGTTTTTGCAACTCTGGAACCGAAGCCGTAATGGCCGCTATTCGTATCGCCAGGGCA
            ACAACAGGTCGGAGTAAAATTGCCCTCTTTGAAGGCTCCTATCATGGACATGCGGACGGA
            ACCCTTTTTAGGAACCAAATTATTGATAACCAACTCCACTCTTTTCCCCTAGCTCTAGGC
            GTTCCCCCCAGCCTTAGTTCCGATGTGGTGGTATTGGACTATGGCAGTGCGGAAGCTCTG
            AACTATTTACAAACCCAGGGGCAGGATTTAGCGGCGGTCTTAGTAGAACCAATTCAAAGT
            GGCAATCCTCTACTCCAACCCCAACAATTTCTCCAAAGTCTGCGACAAATTACCAGTCAA
            ATGGGCATTGCCCTGATTTTTGATGAAATGATTACGGGTTTTCGATCGCACCCAGGGGGA
            GCGCAAGCTTTATTTGGAGTACAGGCGGATATTGCCACCTATGGCAAAGTAGTTGCGGGA
            GGAATGCCCATTGGAGTTATTGCAGGTAAGGCCCATTATCTGGACAGCATTGACGGGGGA
            ATGTGGCGTTATGGCGATAAATCCTATCCTGGGGTGGACAGAACCTTTTTTGGGGGAACC
            TTTAATCAGCATCCGTTAGCAATGGTAGCGGCTAGGGCTGTCCTGACCCATTTAAAGGAG
            CAGGGGCCAGGTCTGCAACAACAATTAACTGAACGCACTGCGGCCTTAGCCGATACACTG
            AATCATTATTTTCAAGCCGAAGAAGTTCCTATTAAAATCGAACAGTTTAGTTCTTTCTTC
            CGGTTTGCCCTCTCTGGCAATTTGGATTTACTTTTCTATCACATGGTAGAAAAAGGTATT
            TATGTCTGGGAATGGCGTAAACATTTTCTTTCAACCGCCCATACGGAAGCCGATCTTGCC
            CAATTTGTCCAAGCGGTTAAGGATAGCATCACAGAATTGCGT
SEQ ID      GGGGGGGATCAAGTCCCTCTCACCGAAGCCCAACGACAACTGTGGATTTTGGCTCAATTA
NO. 31 C    GGAGACAACGGCTCTGTGGCCTATAACCAATCAGTGACATTGCAATTAAGTGGCCCATTA
nucl acid   AATCCCGTCGCAATGAATCAAGCTATTCAACAAATCAGCGATCGCCATGAAGCGTTACGA
            ACCAAAATTAATGCCCAGGGAGATAGTCAAGAAATCCTGCCCCAGGTCGAAATTAACTGC
            CCTATCTTAGACTTCAGTCTTGACCAAGCTTCGGCCCAACAGCAAGCAGAACAATGGTTA
            AAGGAAGAAAGTGAAAAACCCTTTGATTTGAGCCAGGGTTCTCTCGTGCGTTGGCATCTA
            CTCAAATTAGAACCAGAATTACATTTGTTAGTATTAACGGCCCATCACATTATCAGTGAC
            GGTTGGTCAATGGGGGTAATCCTTCGGGAATTAGGAGAGTTATATTCAGCCAAATGTCAG
            GGTGTTACGGCTAATCTTAAAACCCCAAAACAGTTTCGAGAATTGATTGAATGGCAAGC
            CAGCCAAGCCAAGGGGAAGAACTGAAAAAACAGCAAGCCTATTGGTTAGCAACCCTTGCC
            GATCCCCCTGTTTTGAATTTACCCACTGACAAACCTCGTCCAGCTTTACCCAGTTACCAA
            GCTAATCGTCGAAGTCTAACTTTAGATAGCCAATTTACAGAAAAACTAAAGCAATTTAGT
            CGTAAACAGGGCTGTACCTTGCTGATGACCCTGTTATCGGTTTATAACATTCTCGTTCAT
            CGTTTGACGGGACAGGATGATATTCTGGTGGGTCTGCCAGCCTCTGGACGGGGGCTTTTA
            GATAGTGAAGGTATGGTGGGTTATTGCACCCATTTTTTACCAATTCGCAGTCAATTAGCA
SEQ ID      ACTTACAGTGAATTAAATTGTCGAGCCAATCAGTTAGCACATTATTTACAAAAATTAGGA
NO. 32 A    GTTGGGCCAGAGGTCTTAGTCGGTATTTTGGTCGAACGTTCTTTAGAAATGATTGTCGGA
nucl acid   TTGTTAGGGATTCTCAAGGCTGGGGGAGCCTATGTACCTCTTGATCCTGACTATCCCCCT
            GAACGTCTTCAATTTATGTTAGAAGATAGTCAATTTTTTCTCCTCTTAACCCAACAGCAT
            TTACTGGAATCTTTTGCTCAGTCTTCAGAAACGGCTACTCCCAAGATTATTTGTTTGGAT
            AGCGACTACCAAATTATTTCCCAGGCAAAGAATATTAATCCCGAAAATTCAGTCACAACG
            AGTAATCTTGCCTATGTAATTTATACCTCTGGTTCGACAGGTAAACCGAAGGGCGTGATG
            AATAATCATGTTGCTATTAGTAATAAATTGTTATGGGTACAAGACACTTATCCTCTAACC
            ACAGAAGACTGTATTTTACAAAAAACTCCCTTTAGTTTTGATGTTTCAGTGTGGGAATTA
            TTCTGGCCCCTACTAAACGGAGCGCGTTTGGTTTTTGCCAAGCCGAATGGCCATAAAGAT
            GCCAGTTACTTAGTCAATCTGATTCAAGAGCAACAAGTAACAACGCTACATTTTGTGTCT
            TCTATGCTACAGCTTTTTCTGACAGAAAAAGACGTAGAAAAATGTAATAGTCTTAAACGA
            GTCATTTGTAGTGGTGAAGCCCTTTCTTTAGAGCTTCAAGAACGTTTTTTTGCTCGTTTA
            GTCTGTGAATTACACAATCTTTATGGACCGACAGAAGCCGCTATTCATGTCACATTTTGG
            CAATGTCAATCAGATAGCAATTTGAAAACAGTACCCATTGGTCGGCCGATCGCTAATATC
            CAAATTTACATTTTAGACTCTCATCTTCAGCCAGTACCTATTGGAGTAATCGGAGAATTG
            CACATTGGTGGGGTTGGTTTGGCGCGGGGTTATTTAAACAGGCCTGAGTTAACGGCGGAG
            AAATTTATTGCAAATCCGTTTGCTTCCCTTGATCCCCCCCTAACCCCCCTTGATAAGGGG
            GGAGATGAGAGCTATAAAACTTTTAAAAAGGGGGGAGAGCAACCATCAAGATTGTATAAA
            ACGGGAGATTTAGCTCGTTATTTACCCGATGGCAAGATTGAGTATCTAGGGCGCATTGAT
            AATCAGGTAAAAATTCGCGGTTTCCGGATTGAATTGGGGGAAATTGAAGCGGTTTTGCTA
            TCCCATCCCCAGGTACGAGAAGCGGTCGTT
SEQ ID      GAGGCGATCGCCGCTATTTTTGGTCAAGTTTTAAAACTGGAAAAAGTGGGAATTTATGAT
NO. 33 T    AACTTTTTTGAGATCGGCGGTAATTCTTTGCAAGCCACTCAAGTTATTTCACGCTTACGA
nucl acid   GAAAGTTTTGCCCTAGAGTTGCCCTTGCGTCGCCTGTTTGAACAACCGACTGTGGCGGAT
            TTGGCTTTAGCCGTA
SEQ ID      CCTCGTGATGGCCAATTACCCCTCTCCTTTGCCCAGTCGCGACTCTGGTTCTTGTATCAA
NO. 34 C    TTAGAAGGAGCCACGGGAACCTATAACATGACAGGGGCCTTGAGTTTAAGCGGGCCTCTT
2 nucl      CAGGTCGAAGCCCTCAAACAAGCCCTAAGAACTATCATTCAACGCCATGAGCCATTGCGT
acid        ACCAGTTTCCAATCGGTTGACGGGGTTCCAGTGCAGGTGATTAATCCCTATCCTGTTTGG
            GAATTAGCGATGGTTGATTTGACAGGAAAGGAGACAGAAGCAGAAAAATTGGCCTATCAG
            GAATCCCAAACCCCGTTTGATTTGACCAATAGTCCTTTGTTGAGGGTAACGCTCCTCAAA
            TTACAGCCAGAAAAGCATATTTTATTAATTAATATGCACCATATTATTTCCGATGGCTGG
            TCAATCGGTGTTTTTGTTCGTGAATTGTCCCATCTCTATAGGGCTTTTGTGGCGGGTAAA
            GAACCAACTTTACCGATTTTACCAATTCAGTATGCGGATTTTGCCGTTTGGCAGCGAGAG
            TGGTTACAGGGTAAGGTTTTAGCGGCTCAATTGGAATATTGGAAGCGACAATTGGCAGAT
            GCTCCTCCTCTGCTGGAACTGCCCACTGATCGCCCTCGTCCCGCAATCCAAACCTTTCAA
            GGCAAGACAGAAAGATTTGAGCTAGATAGGAAACTGACCCAAGAATTAAAGGCATTAAGT
            CAACAGTCGGGTTGTACTTTATTTATGACTTTGTTGGCCGCTTTTGGGGTGGTTTTATCC
            CGTTATAGTGGCCAGACTGATATCGTCATTGGTTCGGCGATCGCCAACCGTAATCGCCAA
            GACATTGAGGGGTTAATTGGCTTTTTTGTTAACACTTTGGCGTTGAGGTTAGATTTATCA
SEQ ID      ACCTATGGAGAATTAAACCATCGCGCCAATCAATTAGCTCACTATCTTCAGTCGTTAGGA
NO. 35 A    GTCACCAAAGAACAAATCGTCGGGGTTTATCTGGAACGTTCCCTTGAAATGGCGATCGGA
2 nucl      TTTTTAGGTATTCTCAAAGCAGGAGCCGCCTATCTCCCCATTGATCCTGAATATCCCTCA
acid        GTACGCACCCAATTTATTCTCGAAGATACCCAACTTTCGCTTCTCTTAACTCAGGCAGAA
            CTGGCAGAAAAACTGCCCCAGACTCAAAACAAAATTATCTGTCTAGATCGGGACTGGCCA
            GAAATTACCTCCCAACCCCAGACAAACCTAGACCTAAAGATAGAACCTAATAACCTAGCC
            TATTGCATCTATACTTCTGGTTCCACAGGACAACCCAAAGGAGTACTGATTTCCCATCAA
            GCCCTACTCAACTTAATTTTCTGGCATCAACAAGCGTTTGAGATTGGCCCCTTACATAAA
            GCGACCCAAGTGGCAGGCATTGCTTTCGATGCAACGGTTTGGGAATTGTGGCCCTATCTG
            ACCACAGGAGCCTGTATTAATCTGGTTCCCCAAAATATTCTGCTCTCACCGACGGATTTA
            CGGGATTGGTTGCTTAACCGAGAAATTACCATGAGTTTTGTGCCAACTCCTTTAGCTGAA
            AAATTATTATCCTTGGATTGGCCTAACCATTCTTGTCTAAAAACCCTGTTACTGGGAGGT
            GACAAACTTCATTTTTATCCTGCTGCGTCCCTTCCCTTTCAGGTCATTAACAACTATGGC
            CCAACGGAAAATACAGTGGTTGCGACCTCTGGACTGGTCAAATCATCTTCATCTCATCAC
            TTTGGAACTCCGACTATTGGTCGTCCCATTGCCAACGTCCAAATCTATTTATTAGACCAA
            AACCTACAACCTGTCCCCATTGGTGTACCAGGAGAATTACATTTAGGTGGGGCGGGTTTA
            GCGCAGGGCTATCTCAATCGTCCTGAGTTAACGGCTGAAAAATTTATTGCCAATCCCTTT
            GATCCCCCCCTAACCCCCCTTGATAAGGGGGGAGAAGAACCCTCAAAACTCTATAAAACG
            GGAGACTTAGCCCGTTATTTACCCGATGGCAATGTAGAATTTTTGGGACGTATTGACAAT
            CAGGTAAAAATTCGGGGTTTTCGCATCGAAACTGGGGAAATCGAAGCCGTTTTAAGTCAA
            TATTTCCTATTAGCTGAAAGTGTAGTC
SEQ ID      GCTCAACTGACTCAAATTTGGAGTGAAGTTTTGGGACTGGAACGCATTGGCGTTAAGGAC
NO. 36 T    AACTTTTTTGAATTGGGAGGACATTCTCTTTTGGCTACCCAGGTTTTATCAAGAATTAAT
2 nucl      TCAGCCTTTGGACTTGATCTTTCTGTGCAAATTATGTTTGAATCACCAACGATCGCGGGC
acid        ATTGCGGGTTATATT
SEQ ID      GCTAGAGACGGTCATTTACCCCTGTCTTTTGCTCAACAACGTTTATGGTTTTTACATTAT
NO. 37 C    CTTTCCCCTGATAGTCGTTCCTACAATACCCTGGAAATATTGCAAATTGATGGGAATCTC
3 nucl      AATCTGACTGTGCTAGAGCAGAGTTTGGGGGAATTAATTAACCGCCATGAAATTTTTAGA
acid        ACAACATTCCCCACTGTTTCAGGGGAACCGATTCAGAAAATTGCACTTCCTAGTCGTTTT
            CAGTTAAAAGTTGATAATTATCAAGATTTAGACGAAAATGAACAATCAGCTAAAATTCAA
            CAAGTAGCAGAATTGGAAGCAGGACAAGCTTTTGATTTAACGGTGGGGCCACTGATTCAG
            TTTAAGCTATTGCAATTGAGTCCCCAGAAGTCGGTGCTGCTGTTGAAAATGCACCATATT
            ATCTATGATGGCTGGTCTTTTGGGATTCTGATTCGGGAATTATCGGCTCTATACGAAGCA
            TTTTTAAAGAACTTAGCCAATCCTCTCCCTGCGTTGTCTATTCAGTATGCAGATTTTGCG
            GTTTGGCAACGTCAATATCTCTCAGGTGAGGTCTTAGATAAACAACTCAATTATTGGCAA
            GAACAGTTAGCAACAGTCTCTCCTGTTCTTACTTTACCAACGGATAGACCCCGTCCGGCG
            ATACAAACTTTTCAGGGAGGAGTTGAGCGTTTTCAACTGGATCAAAATGTCACTCAAGGT
            CTTAAAAAGTTAGGTCAAGATCAGGTTGCAACCCTGTTTATGACGTTGTTGGCCGGTTTC
            GGCGTTTTGCTATCTCGTTATAGTGGTCAATCTGATCTGATGGTGGGTTCTCCGATCGCT
            AATCGTAATCAAGCAGCGATCGAACCTTTAATTGGCTTTTTTGCTAACACTTTGGCTTTA
            AGAATTAATTTATCA
SEQ ID      ACATACACTGAATTAAACCATCGCGCTAATCAGTTAGCCCATTATTTACAAACTTTAGGC
NO. 38 A    GTGGGAGCAGAAGTCTTAGTCGGTATTTCCCTAGAACGTTCTTTAGAGATGATTATCGGC
3 nucl      TTATTAGGGATTCTCAAGGTAGGTGGTGCTTATCTTCCTCTTGATCCAGACTATCCCACT
acid        GAGCGTCTTCAGTTGATGTTAGAAGACAGTCAAGTTCCTTTTTTGATTACCCACAGTTCT
            TTATTAGCAAAATTGCCTCCCTCTCAAGCAACTCTGATTTGTTTAGATCATATCCAAGAG
            CAGATTTCTCAATATTCTCCAGATAATCTTCAATGTCAGTTAACTCCTGCCAATTTAGCT
            AACGTTATTTATACCTCTGGCTCTACGGGTAAGCCTAAAGGGGTGATGGTTGAACATAAA
            GGTTTAGTTAACTTAGCTCTTGCTCAAATTCAATCTTTTGCAGTCAACCATAACAGTCGT
            GTGCTGCAATTTGCTTCTTTTAGTTTTGATGCTTGTATTTCAGAAATTTTGATGACCTTT
            GGTTCTGGAGCGACGCTTTATCTTGCACAAAAAGATGCTTTATTGCCAGGTCAGCCATTA
            ATTGAACGGTTAGTAAAGAATGGAATTACTCATGTGACTTTGCCGCCTTCAGCTTTAGTG
            GTTTTACCCCAGGAACCGTTACGCAACTTAGAAACCTTAATTGTGGCGGGTGAGGCTTGT
            TCTCTTGATTTAGTGAAACAATGGTCAATCGATAGAAACTTTTTCAATGCCTATGGGCCA
            ACGGAAGCGAGTGTTTGTGCCACTATTGGACAATGTTATCAAGATGATTTAAAGGTGACG
            ATTGGTAAGGCGATCGCCAATGTCCAAATTTATATTTTAGATGCCTTTTTACAGCCGGTG
            CCGGTGGGAGTGTCAGGAGAGTTATACATTGGTGGAGTTGGGGTGGCAAGGGGCTATTTA
            AATCGTCCTGAATTAACCCAAGAAAAATTTATTGCTAATCCTTTTAGTAACGACCCAGAT
            TCTCGGCTCTATAAAACTGGCGACTTAGCGCGTTATTTACCCGATGGTAATATTGAATAT
            TTAGGACGCATTGACAATCAGGTAAAAATTCGCGGTTTTCGCATTGAGTTAGGAGAAATT
            GAAGCGGTTCTGAGTCAATGTCCCGATGTGCAAAATACGGCGGTG
SEQ ID      GAAATTCTGGCTCAAATATGGGGGCAAGTTCTCAAGATAGAAAGAGTCAGCAGAGAAGAT
NO. 39 T    AATTTCTTTGAATTGGGGGGGCATTCCCTTTTAGCTACCCAGGTAATGTCCCGTCTGCGT
3 nucl      GAAACTTTTCAAGTCGAATTACCTTTGCGTAGTCTCTTTACCGCTCCCACTATTGCTGAA
acid        TTGGCCCTAACAATT
SEQ ID      AACGACAGTGCTAACCTCCCGTTATCTTTTGCTCAACAACGTTTATGGTTTCTGGATCAA
NO. 40 C    TTAGAACCTAACAGCGCCTTTTATCATGTAGGGGGAGCCGTAAGACTAGAAGGAACATTA
4 nucl      AATATTACTGCCTTAGAGCAAAGCTTAAAAGAAATTATTAATCGTCATGAAGCTTTACGC
acid        ACAAATTTTATAACGATTGATGGTCAAGCCACTCAAATTATTCACCCTACTATTAATTGG
            CGATTGTCTGTTGTTGATTGTCAAAATTTAACCGACACTCAATCTCTGGAAATTGCGGAA
            GCTGAAAAGCCCTTTAATCTTGCTCAAGATTGCTTATTTCGTGCTACTTTATTCGTGCGA
            TCACCGCTAGAATATCATCTACTCGTGACCATGCACCATATTGTTAGCGATGGCTGGTCA
            ATTGGAGTATTTTTTCAAGAACTAACTCATCTTTACGCTGTCTATAATCAGGGTTTACCC
            TCATCTTTAACGCCTATTAAAATACAATATGCTGATTTTGCGGTCTGGCAACGGAATTGG
            TTACAAGGTGAAATTTTAAGTAATCAATTGAATTATTGGCGCGAACAATTAGCAAATGCT
            CCTGCTTTTTTACCTTTACCGACAGATAGACCTAGGCCCGCAATCCAAACTTTTATTGGT
            TCTCATCAAGAATTTAAACTTTCTCAGCCATTAAGCCAAAAATTGAATCAACTAAGTCAG
            AAGCATGGAGTGACTTTATTTATGACTCTCCTGGCTGCTTTTGCTACCTTACTTTACCGT
            TATACAGGACAAGCAGATATTTTAGTTGGTTCTCCTATTGCTAACCGTAATCGTAAGGAA
            ATTGAGGGATTAATCGGCTTTTTTGTTAATACATTAGTTCTGAGATTGAGTTTAGAT
SEQ ID      ACCTATGCTGAATTAAATCATCAAGCTAATCAGTTAGTCCATTACTTACAAACTTTAGGA
NO. 41 A    ATTGGGCCAGAGGTCTTAGTCGCTATTTCAGTAGAACGTTCTTTAGAAATGATTATCGGC
4 nucl      TTATTAGCCATTCTCAAGGCGTGTGGTGCTTATCTCCCTCTTGCTCCTGACTATCCCACT
acid        GAGCGTCTTCAGTTCATGTTAGAAGATAGTCAAGCTTCTTTTTTGATTACCCACAGTTCT
            TTATTAGAAAAATTGCCTTCTTCTCAAGCGACTCTAATTTGTTTAGATCACATCCAAGAG
            CAGATTTCTCAATATTCTCCCGATAATCTTCAAAGTGAGTTAACTCCTTCCAATTTGGCT
            AACGTTATTTACACCTCTGGCTCTACGGGTAAGCCTAAAGGGGTGATGGTTGAACATCGG
            GGCTTAGTTAACTTAGCGAGTTCTCAAATTCAATCTTTTGCAGTCAAAAATAACAGTCGT
            GTACTGCAATTTGCTTCCTTTAGTTTTGATGCTTGTATTTCAGAAATTTTGATGACCTTT
            GGTTCTGGAGCGACTCTTTATCTTGCTCAAAAAAATGATTTATTGCCAGGTCAGCCATTA
            ATGGAAAGGTTAGAAAAGAATAAAATTACCCATGTTACTTTACCCCCTTCAGCTTTAGCT
            GTTTTACCAAAAAAACCGTTACCCAACTTACAAACTTTAATTGTGGCGGGTGAGGCTTGT
            CCTCTGGATTTAGTCAAACAATGGTCAGTCGGTAGAAACTTTTTCAATGCCTATGGCCCG
            ACAGAAACGAGTGTTTGTGCCACGATTGGACAATGTTATCAAGATGATTTAAAGGTCACG
            ATTGGTAAGGCGATCGCTAATGTCCAAATTTATATTTTGGATGCCTTTTTACAACCAGTA
            CCCATCGGAGTACCAGGGGAATTATACATTGGTGGAGTCGGAGTTGCGAGGGGTTATCTA
            AATCGTCCTGAATTAACGGCGGAAAGATTTATTCCTAATCCTTTTGATCCCCCCCTAACC
            CCCCTTAAAAAGGGGGGAGATAAGAGCTATGAAACTTTTAAAAAGGGGGAAGAGCAACCA
            TCAAAACTCTATAAAACGGGAGATTTAGCTCGTTATTTACCCGATGGCAATATTGAATAT
            TTAGGACGCATTGACAATCAGGTAAAAATTCGCGGTTTTCGCATTGAGTTAGGAGAAATT
            GAAGCGGTTCTGAGTCAATGTCCCGATGTGCAAAATACGGCGGTG
SEQ ID      TTACAATTAGCTCAAATCTGGTCAGAGATTTTAGGCATTAATAATATTGGTATTCAGGAA
NO. 42 T    AACTTCTTTGAATTAGGCGGTCATTCTTTATTAGCAGTCAGTCTGATCAATCGTATTGAA
4 nucl      CAAAAGTTAGATAAACGTTTACCATTAACCAGTCTTTTTCAAAATGGAACCATAGCAAGT
acid        CTAGCTCAATTACTAG
SEQ ID      ACTCCATTTTTTGCTGTTCATCCCATTGGTGGTAATGTGCTATGTTATGCCGATTTAGCT
NO. 43      CGTAATTTAGGAACGAAACAGCCGTTTTATGGATTACAATCATTAGGGCTAAGTGAATTA
TE nucl     GAAAAAACTGTAGCCTCTATTGAAGAAATGGCGATGATTTATATTGAAGCAATACAAACT
acid        GTTCAAGCCTCTGGTCCCTACTATTTAGGAGGTTGGTCAATGGGAGGAGTGATAGCTTTT
            GAAATCGCCCAACAATTATTGACCCAAGGTCAAGAAGTTGCTTTACTGGCTTTAATAGAT
            AGTTATTCTCCCAGTTTACTTAATTCAGTTAATAGGGAGAAAAATTCTGCTAATTCCCTG
            ACAGAAGAATTTAATGAAGATATCAATATTGCCTATTCTTTCATCAGAGACTTAGCAAGT
            ATATTTAATCAAGAAATCTCTTTCTCTGGGAGTGAACTTGCTCATTTTACATCAGACGAA
            TTACTAGACAAGTTTATTACTTGGAGTCAAGAGACGAATCTTTTGCCGTCAGATTTTGGG
            AAGCAGCAGGTTAAAACCTGGTTTAAAGTTTTCCAGATTAATCACCAAGCTTTGAGCAGC
            TATTCTCCCAAGACGTATCTGGGTAGAAGTGTTTTCTTAGGAGCGGAAGACAGTTCTATT
            AAAAATCCTGGTTGGCATCAA
SEQ ID      AGCGGGTCTCAAGACCAAAAAACGATACAGTTTAGCCTCTACTACTTTGGTAGCTATGAA
NO. 44      GCGGAATTTAACCCGAATAAATATAACTTACTGTTTGAAGGAGCTAAATTTGGCGATCGC
MO nucl     GCTGGTTTTACGGCCCTTTGGATTCCTGAACGTCATTTCCACGCTTTTGGTGGTTTTTCT
acid        CCCAATCCTTCGGTTTTGGCGGCGGCTTTAGCACGGGAAACCAAACAGATTCAACTGCGA
            TCAGGCAGTGTGGTTTTACCGCTACATAATTCCATCCGAGTCGCCGAAGAATGGGCAGTG
            GTGGACAATCTTTCCCAGGGCCGCGTTGGTATTGCTTTTGCATCGGGTTGGCATCCCCAG
            GATTTTGTCTTGGCTCCCCAGTCCTTTGGCCAACATCGGGAATTGATGTTCCAAGAAATT
            GAAACCGTCCAGAAACTTTGGCGAGGGGAAGCGATCACCGTGCCAGACGGAAAGGGTCAA
            AGGGTAGAGGTTAAAACCTATCCCCAACCGATGCAGTCCCAGTTACCCAGCTGGATTACT
            ATTGTCAATAATCCCGATACCTATATCAGAGCAGGGGCGATCGGTGCTAATATCCTTACC
            AATCTGATGGGGCAAAGCGTGGAAGATTTAGCCCGTAATATTGCGCTATATCGTCAATCT
            TTGGCAGAGCATGGTTATGATCCCGCGTCGGGAACGGTGACAGTTCTCCTGCATACTTTT
            GTTGGCAAGGATTTAGAACAAGTTCGAGAACAGGCTCGCCAACCCTTTGGGCAATACCTC
            ACCTCCTCTGTCGGACTCTTGCAGAACATGGTCAAGAGCCAGGGCATGAAAGTGGATTTT
            GAACAATTAAGAGACGAAGATCGGGACTTTCTCCTCGCTTCTGCCTATAAACGCTATACA
            GAAACCAGTGCTTTAATTGGCACACCCGAATCCTGTCGTCAAATTATTGATCATTTGCAG
            TCCATCGGTGTGGATGAAGTGGCTTGTTTTATTGATTTTGGGGTAGATGAACAAACAGTT
            TTGGCCAATTTACCCTATCTCCAGTCCCTAAAAGACTTATATCAA
SEQ ID      ATTGATCCCCCCCTAACCCCCCTTGATAAGGGGATTGATCCCCCCCTAACCCCCCTTGAT
NO. 45      AAGGGGATTGATCCCCCCCTAACCCCCCTTGATAAGGGG
SP 1 nucl
acid
SEQ ID      CCTTATCAAGGGGGGTTAGGGGGGGATCAATCCCCTTATCAAGGGGGGTTAGGGGGGGAT
NO. 46      CAATCCCCTTATCAAGGGGGGTTAGGGGGTGATCAATCCCCTTATCAAGGGGGGTTAGGG
SP 2 nucl   GGTGATCAATCCCCTTATCAAGGGGGGTTAGGGGGGGATCAATCCCCTTATCAAGGAGAG
acid        TTAGGGGGGGATCAATCCCCTTATCAAGGGGGGTTAGGGGGGGATCAAGTC
SEQ ID      CCTGCTTCAGAAATGCGAGAGTGGGTCGAAAACACTGTTAGTCGCATCTTGGCTTTCCAA
NO. 47      CCAGAACGCGGTTTAGAAATTGGTTGTGGTACAGGTTTGTTACTCTCCAGGGTAGCAAAG
MT nucl     CATTGTCTTGAATATTGGGCAACGGATTATTCCCAAGGGGCGATCCAGTATGTTGAACGG
acid        GTTTGCAATGCCGTTGAAGGTTTAGAACAGGTTAAATTACGCTGTCAAATGGCAGATAAT
            TTTGAAGGTATTGCCCTACATCAATTTGATACCGTCGTCTTAAATTCGATTATTCAGTAT
            TTTCCCAGTGTGGATTATCTGTTACAGGTGCTTGAAGGGGCGATCAACGTCATTGGCGAG
            CGAGGTCAGATTTTTGTCGGGGATGTGCGGAGTTTACCCCTATTAGAGCCATATCATGCG
            GCTGTGCAATTAGCCCAAGCTTCTGACTCGAAAACTGTTGAACAATGGCAACAACAGGTG
            CGTCAAAGTGTAGCAGGTGAAGAAGAACTGGTCATTGATCCCACATTGTTCCTGGCTTTA
            AAACAACATTTTCCGCAAATTAGCTGGGTAGAAATTCAACCGAAACGGGGTGTGGCTCAC
            AATGAGTTAACTCAATTTCGCTATGATGTCACTCTCCATTTAGAGACTATCAATAATCAA
            GCATTATTGAGCGGCAATCCAACGGTAATTACCTGGTTAAATTGGCAACTTGACCAACTG
            TCTTTAACACAAATTAAAGATAAATTATTAACAGACAAACCTGAATTGTGGGGAATTCGT
            GGTATTCCTAATCAGCGAGTTGAAGAGGCTCTAAAAATTTGGGAATGGGTGGAAAATGCC
            CCTGATGTTGAAACGGTTGAACAACTCAAAAAACTTCTCAAACAACAAGTAGATACTGGT
            ATTAATCCTGAACAGGTTTGGCAATTAGCTGAGTCTCTCGGTTACACCGCTCACCTTAGT
            TGGTGGGAAAGTAGTCAAGACGGTTCCTTTGATGTCATTTTTCAGCGGAATTCAGAAGCG
            GAGGACTCAAAAAAATTAACCCTTTCAAAACTTGCTTTCTGGGATGAAAAACCCTTTAAA
            ATAAAGCCCTGGAGTGACTATACTAACAACCCTCTGCGCGGTAAGTTAGTCCAAAAATTA
            ATTCCT
SEQ ID      ATGACAAATTATGGCAAATCTATGTCTCATTACTATGATCTAGTGGTAGGACATAAAGGT
NO. 48      TATAACAAAGATTACGCCACTGAAGTAGAATTCATTCACAATTTAGTTGAGACTTACACA
MT 2        ACTGAAGCCAAATCTATCCTATACTTGGGCTGTGGTACGGGTTATCATGCCGCTCTTTTA
nucl acid   GCACAGAAAGGGTATTCTGTACATGGTGTTGATCTCAGTGCTGAAATGTTAGAGCAGGCT
            AAAACTCGCATTGAAGATGAAACAATAGCTTCTAATCTGAGTTTTTCTCAAGGAAATATT
            TGTGAAATCCGTTTAAATCGTCAGTTTAATGTTGTTCTTGCTCTATTTCATGTGGTTAAC
            TATCAAACGACCAATCAAAATTTACTGGCAACGTTTGCAACGGTTAAAAACCATTTAAAA
            GCTGGGGGGATTTTTATTTGTGATGTGTCCTATGGGTCTTACGTACTGGGGGAATTTAAG
            AGTCGGCCTACGGCATCAATATTGCGTTTAGAGGATAATTCCAATGGTAACGAAGTAACC
            TATATTAGTGAACTAAATTTTTTAACCCATGAAAATATAGTGGAAGTTACTCACAATTTA
            TGGGTAACAAATCAAGAAAATCAACTTCTAGAGAATTCACGGGAAACACATCTTCAGCGC
            TATCTTTTCAAGCCTGAAGTTGAATTGTTGGCTGATGCTTGTGAACTAACTGTTCTTGAT
            GCGATGCCCTGGCTTGAACAACGTCCTTTGACAAACATTCCTTGTCCTTCAGTTTGTTTT
            GTTATTGGGCATAAAACAACCCATTCAGCTTAA
SEQ ID      CCGACCTGTGATAAACAATTC
NO. 49
Primer A
SEQ ID      CKNCCDGTDATRAANARYTC
NO. 50
Primer B
SEQ ID      TTCAATATCCTGGGGATA
NO. 51
Primer C
SEQ ID      YTCDATRTCYTGNGGRTA
NO. 52
Primer D
SEQ ID      CGTTGGTTACAGGCCCTTTCT
NO. 53
Primer E
SEQ ID      MGNTGGYTNCARGCNYTNWS
NO. 54
Primer F
SEQ ID      TTAGACTTAAGCCATTGG
NO. 55
Primer G
SEQ ID      YTNGAYYTNWSNCAYTGG
NO. 56
Primer H
SEQ ID      CATAGAAGAATCGAGACCATATTC
NO. 57
Primer I
SEQ ID      CATNSWNSWRTCNARNCCRTAYTC
NO. 58
Primer J
SEQ ID      MTTQTASSANALASFNQFLRDVKAIAQPYWYPTVSNKRSFSEVIRSWGMLSLLIFLIVGL
NO. 59      VAVTAFNSFVNRRLIDVIIQEKDASQFASTLTVYAIGLICVTLLAGFTKDIRKKIALDWY
ABC         QWLNTQIVEKYFSNRAYYKINFQSDIDNPDQRLAQEIEPIATNAISFSATFLEKSLEMLT
Transporter FLVVVWSISRQIAIPLMFYTIIGNFIAAYLNQELSKINQAQLQSKADYNYALTHVRTHAE
            SIAFFRGEKEEQNIIQRRFQEVINDTKNKINWEKGNEIFSRGYRSVIQFFPFLVLGPLYI
            KGEIDYGQVEQASLASFMFASALGELITEFGTSGRFSSYVERLNEFSNALETVTKQAENV
            STITTIEENHFAFEHVTLETPDYEKVIVEDLSLTVQKGEGLLIVGPSGRGKSSLLRAIAG
            LWNAGTGRLVRPPLEEILFLPQRPYIILGTLREQLLYPLTNSEMSNTELQAVLQQVNLQN
            VLNRVDDFDSEKPWENILSLGEQQRLAFARLLVNSPSFTILDEATSALDLTNEGILYEQL
            QTRKTTFISVGHRESLFNYHQWVLELSADSSWELLSVQDYRLKKAGEMFTNASSNNSITP
            DITIDNGSEPEIVYSLEGFSHQEMKLLTDLSLSSIRSKASRGKVITAKDGFTYLYDKNPQ
            ILKWLR
SEQ ID      ATGACAACCCAAACAGCTTCTAGTGCCAATGCCCTTGCTTCCTTTAACCAATTTTTAAGG
NO. 60      GATGTAAAGGCGATCGCCCAACCCTATTGGTATCCCACTGTATCAAATAAAAGAAGCTTT
ABC         TCTGAGGTTATTCGTTCCTGGGGAATGCTATCACTGCTTATCTTTTTGATTGTGGGATTA
Transporter GTCGCCGTCACGGCTTTTAATAGTTTTGTTAATCGTCGTTTAATTGATGTCATTATTCAA
Nucl acid   GAAAAAGATGCGTCTCAATTTGCCAGTACATTAACTGTCTATGCGATCGGATTAATCTGT
            GTAACGCTGCTGGCAGGGTTCACTAAAGATATTCGCAAAAAAATTGCCCTAGATTGGTAT
            CAATGGTTAAACACCCAGATTGTAGAGAAATATTTTAGTAATCGTGCCTATTATAAAATT
            AACTTTCAATCTGACATTGATAACCCCGATCAACGTCTAGCCCAGGAAATTGAACCGATC
            GCCACAAACGCCATTAGTTTCTCGGCCACTTTTTTGGAAAAAAGTTTGGAAATGCTAACT
            TTTTTAGTGGTAGTTTGGTCAATTTCTCGACAGATTGCTATTCCGCTAATGTTTTACACG
            ATTATCGGTAATTTTATTGCCGCCTATCTAAATCAAGAATTAAGCAAGATCAATCAGGCA
            CAACTGCAATCAAAAGCAGATTATAACTATGCCTTAACCCATGTTCGGACTCATGCGGAA
            TCTATTGCTTTTTTTCGGGGAGAAAAAGAGGAACAAAATATTATTCAGCGACGTTTTCAG
            GAAGTTATCAATGATACGAAAAATAAAATTAACTGGGAAAAAGGGAATGAAATTTTTAGT
            CGGGGCTATCGTTCCGTCATTCAGTTTTTTCCTTTTTTAGTCCTTGGCCCTTTGTATATT
            AAAGGAGAAATTGATTATGGACAAGTTGAGCAAGCTTCATTAGCTAGTTTTATGTTTGCA
            TCGGCCCTGGGAGAATTAATTACAGAATTTGGTACTTCAGGACGTTTTTCTAGTTATGTA
            GAACGTTTAAATGAATTTTCTAATGCCTTAGAAACTGTGACTAAACAAGCCGAGAATGTC
            AGCACAATTACAACCATAGAAGAAAATCATTTTGCCTTTGAACACGTCACCCTAGAAACC
            CCTGACTATGAAAAGGTGATTGTTGAGGATTTATCTCTTACTGTTCAAAAAGGTGAAGGA
            TTATTGATTGTCGGGCCCAGTGGTCGAGGTAAAAGTTCTTTATTAAGGGCGATCGCCGGT
            TTATGGAATGCTGGCACTGGGCGTTTAGTGCGTCCTCCCCTAGAAGAAATTCTCTTTTTG
            CCCCAACGTCCCTACATTATTTTGGGAACCTTACGCGAACAATTGCTGTATCCTCTAACC
            AATAGTGAGATGAGCAATACCGAACTTCAAGCAGTATTACAACAAGTCAATTTGCAAAAT
            GTGCTAAATCGGGTGGATGACTTTGACTCCGAAAAACCCTGGGAAAACATTCTCTCCCTC
            GGTGAACAACAACGCCTAGCCTTTGCTCGATTGTTAGTGAATTCTCCGAGTTTTACCATT
            TTAGATGAGGCGACCAGTGCCTTAGATTTAACAAATGAGGGGATTTTATACGAGCAATTA
            CAAACTCGCAAGACAACCTTTATTAGTGTGGGTCATCGAGAAAGTTTGTTTAATTACCAT
            CAATGGGTTTTAGAACTTTCTGCTGACTCTAGTTGGGAACTCTTAAGCGTTCAAGATTAT
            CGCCTTAAAAAAGCGGGAGAAATGTTTACTAATGCTTCGAGTAACAATTCCATAACACCC
            GATATTACTATCGATAATGGATCAGAACCAGAAATAGTCTATTCTCTTGAAGGATTTTCC
            CATCAGGAAATGAAACTATTAACAGACCTATCACTCTCTAGCATTCGGAGTAAAGCCAGT
            CGAGGGAAGGTGATTACAGCCAAGGATGGTTTTACCTACCTTTATGACAAAAATCCTCAG
            ATATTAAAGTGGCTCAGAACTTAA

In one embodiment the entire gene cluster is transformed and expressed in a heterologous system. SEQ ID NO. 61 encompasses the genes of said cluster.

1-27260           ATGACTATTAACTATGGTGATCTGCAAGAACCCTTTAATAAATTCTCAACCCTAGTTGAA
Microginin-       TTACTCCGTTATCGGGCAAGCAGTCAACCGGAACGCCTCGCCTATATTTTTCTGCGAGAC
Cluster           GGAGAAATCGAAGAAGCTCGTTTAACCTATGGGGAACTGGATCAAAAGGCTAGGGCGATC
1-1743            GCCGCTTATCTACAATCCTTAGAAGCCGAGGGCGAAAGGGGTTTACTGCTCTATCCCCCA
Adenylation-      GGACTAGATTTTATTTCAGCTTTTTTTGGTTGTTTATATGCGGGAGTCGTTGCCATTCCC
Protein (A*)      GCCTATCCACCCCGACGGAATCAAAACCTTTTGCGTTTACAGGCGATTATTGCCGATTCT
1892-2158         CAAGCCCGATTTACCTTCACCAATGCCGCTCTATTTCCCAGTTTAAAAAACCAATGGGCT
Acyl-Carrier-     AAAGACCCTGAATTAGGAGCAATGGAATGGATTGTTACCGATGAAATTGACCATCACCTC
Protein (ACP)     AGGGAGGATTGGCTAGAACCAACCCTCGAAAAAAACAGTCTCGCTTTTCTACAATACACC
2204-3016         TCTGGTTCAACGGGAACTCCAAAGGGAGTAATGGTCAGTCACCATAATTTGTTGATTAAT
Methyltransferase TCAGCCGATTTAGATCGTGGTTGGGGCCATGATCAAGATAGCGTAATGGTCACTTGGCTA
(MT)              CCGACCTTCCATGATATGGGTCTGATTTATGGGGTTATTCAGCCTTTGTACAAAGGATTT
3464-13123        CTTTGTTACATGATGTCCCCTGCCAGCTTTATGGAACGACCGTTACGTTGGTTACAGGCC
PKS/NRPS (KS-AT-  CTTTCTGATAAAAAAGCAACCCATAGTGCGGCCCCCAACTTTGCCTACGATCTTTGTGTG
ACP-AMT-MO-C-A-T) CGGAAAATTCCCCCTGAAAAACGGGCTACGTTAGACTTAAGCCATTGGTGCATGGCCTTA
13120-17832       AATGGGGCCGAACCCGTCAGAGCGGAGGTACTTAAAAAGTTTGCGGAGGCTTTTCAAGTT
NRPS 2 (C-A-Mt-T) TCTGGTTTCAAAGCCACAGCCCTTTGTCCTGGCTACGGTTTAGCAGAAGCCACCCTGAAA
17836-25194       GTTACGGCGGTTAGTTATGACAGTCCCCCTTACTTTTATCCCGTTCAGGCTAATGCTTTA
NRPS 3 (C-A-T-C-  GAAAAAAATAAGATTGTGGGAGCCACTGAAACCGATACCAATGTGCAGACCCTCGTGGGC
A-T)              TGCGGCTGGACAACGATTGATACTCAAATCGTCATTGTCAATCCTGAAACCCTGAAACCT
25257-27260       TGCTCCCCTGAAATTGTCGGCGAAATTTGGGTATCAGGTTCAACAATCGCCCAAGGCTAT
ABC-Transporter   TGGGGAAAACCTCAAGAGACTCAGGAAACCTTTCAAGCTTATTTGGCAGATACAGGAGCC
(ABC)             GGGCCTTTTCTGCGAACAGGAGACTTGGGCTTCATTAAAGATGGTGAATTGTTTATCACA
                  GGTCGGCTCAAGGAAATTATTCTGATTCGAGGACGCAATAATTATCCCCAGGATATTGAA
                  TTAACCGTCCAAAATAGTCATCCCGCTCTGCGTCCCAGTTGTGGGGCTGCTTTTACCGTT
                  GAAAATAAGGGCGAAGAAAAGCTCGTGGTCGTTCAGGAAGTGGAGCGCACCTGGCTCCGT
                  AAGGTAGATATAGATGAGGTAAAAAGAGCCATTCGTAAAGCTGTTGTCCAGGAATATGAT
                  TTACAGGTTTATGCGATCGCGCTGATCAGGACTGGCAGTTTACCAAAAACCTCTAGCGGT
                  AAAATTCAGCGTCGTAGCTGTCGGGCCAAATTTTTAGAGGGAAGCCTGGAAATTTTGGGC
                  TAAGAAAATTTCTCGATCGGCACTTAATGTGTTAAATTCGTATGTCGATTGAAACTTCGA
                  CCAATTCTTTCTCTCCCCTTAAGTCCATGTCTCTGGATTTGAAAATTCCTTAAACTTTAA
                  CTACATTTCTCAAGAAAGCAAATTGAATCTAATGTCCACAGAAATCCCAAACGACAAAAA
                  ACAACCGACCCTAACGAAAATTCAAAACTGGTTAGTGGCTTACATGACAGAGATGATGGA
                  AGTGGACGAAGATGAGATTGATCTGAGCGTTCCCTTTGATGAATATGGTCTCGATTCTTC
                  TATGGCAGTTGCTTTGATCGCTGATCTAGAGGATTGGTTACGACGAGATTTACATCGCAC
                  CCTGATCTACGATTATCCAACTCTAGAAAAGTTGGCTAAACAGGTTAGTGAACCCTGACA
                  TTTTTATAAAGTTTGTGCTTAAAAATTTTGAGGAAGTTCTAAAATGACAAATTATGGCAA
                  ATCTATGTCTCATTACTATGATCTAGTGGTAGGACATAAAGGTTATAACAAAGATTACGC
                  CACTGAAGTAGAATTCATTCACAATTTAGTTGAGACTTACACAACTGAAGCCAAATCTAT
                  CCTATACTTGGGCTGTGGTACGGGTTATCATGCCGCTCTTTTAGCACAGAAAGGGTATTC
                  TGTACATGGTGTTGATCTCAGTGCTGAAATGTTAGAGCAGGCTAAAACTCGCATTGAAGA
                  TGAAACAATAGCTTCTAATCTGAGTTTTTCTCAAGGAAATATTTGTGAAATCCGTTTAAA
                  TCGTCAGTTTAATGTTGTTCTTGCTCTATTTCATGTGGTTAACTATCAAACGACCAATCA
                  AAATTTACTGGCAACGTTTGCAACGGTTAAAAACCATTTAAAAGCTGGGGGGATTTTTAT
                  TTGTGATGTGTCCTATGGGTCTTACGTACTGGGGGAATTTAAGAGTCGGCCTACGGCATC
                  AATATTGCGTTTAGAGGATAATTCCAATGGTAACGAAGTAACCTATATTAGTGAACTAAA
                  TTTTTTAACCCATGAAAATATAGTGGAAGTTACTCACAATTTATGGGTAACAAATCAAGA
                  AAATCAACTTCTAGAGAATTCACGGGAAACACATCTTCAGCGCTATCTTTTCAAGCCTGA
                  AGTTGAATTGTTGGCTGATGCTTGTGAACTAACTGTTCTTGATGCGATGCCCTGGCTTGA
                  ACAACGTCCTTTGACAAACATTCCTTGTCCTTCAGTTTGTTTTGTTATTGGGCATAAAAC
                  AACCCATTCAGCTTAAATTCTGCTAAAAAAAATCCAACTTACCTTATTCTCTGAAACCAC
                  ACAAGCCATGAATACAATTCAAGATGCCAAGACCGAAAATTACTCAATCTTAAATCAGTC
                  AATTCCAAGACCTCTCAAACTGAGTAATATCCTATTACGATAAGATTTTGCGTTCTCCTT
                  TGTTTGGAATGTCAGCAGAGGAGTCTCTATATTGGCTAGAGAAATGTTTATGTCAAGAGC
                  ATCAGGGCTTCGATGTACAAGTTAAGTATCATCAAAAAATGCTGAAGAATATGTTACGTT
                  TGACCGATAGTTTGGATTATCTATGGCCAGTTAACCGTGAAATGCGGCTCATGAAAGCTG
                  GGGGGTCAATTGAACGGGCGATCACCAATAACATTAAAGCTTTTCTTCAATTTAAAGAAA
                  CTGTAACCGTATTAAATTAGAAAAACCGCAGTGAGGAATTTGAATGGAACCCATCGCAAT
                  TATTGGTCTTGCTTGCCGCTTTCCAGGGGCTGACAATCCAGAAGCTTTCTGGCAACTCAT
                  GCGAAATGGGGTGGATGCGATCGCCGATATTCCTCCTGAACGTTGGGATATTGAGCGTTT
                  CTACGATCCCACACCTGCCACTGCCAAGAAGATGTATAGTCGCCAGGGCGGTTTTCTAAA
                  AAATGTCGATCAATTTGACCCTCAATTTTTCCGAATTTCTCCCCTAGAAGCCACCTATCT
                  AGATCCTCAACAAAGACTGCTACTGGAAGTCACCTGGGAAGCCTTAGAAAATGCTGCCAT
                  TGTGCCTGAAACCTTAGCTGGTAGCCAATCAGGGGTTTTTATTGGTATCAGTGATGTGGA
                  TTATCATCGTTTGGCTTATCAAAGTCCTACTAACTTGACCGCCTATGTGGGTACAGGCAA
                  CAGCACCAGTATTGCGGCTAACCGTTTATCATATCTGTTTGATTTGCGTGGCCCCAGTTT
                  GGCCGTAGATACCGCTTGCTCTTCTTCCCTCGTCGCCGTTCACTTGGCCTGTCAGAGTTT
                  GCAAAGTCAAGAATCGAACCTCTGCTTAGTGGGGGGAGTTAATCTCATTTTGTCGCCAGA
                  GACAACCGTTGTTTTTTCCCAAGCGAGAATGATCGCCCCCGACAGTCGTTGTAAAACCTT
                  TGACGCGAGGGCCGATGGTTATGTGCGCTCGGAAGGCTGTGGAGTAGTCGTACTTAAACG
                  TCTTAGGGATGCCATTCAGGACGGCGATCGCATTTTAGCAGTGATTGAAGGTTCCGCGGT
                  GAATCAGGATGGTTTAAGTAATGGACTCACGGCCCCTAATGGCCCTGCTCAACAGGCGGT
                  GATTCGTCAGGCCCTGGCAAATGCCCAGGTAAAACCGGCCCAGATTAGCTATGTCGAAGC
                  CCATGGCACGGGGACAGAATTGGGGGATCCGATCGAAGTTAAATCTCTGAAAGCGGTTTT
                  GGGTGAAAAGCGATCGCTCGATCAAACCTGTTGGCTCGGTTCTGTGAAAACCAACATTGG
                  TCATTTAGAAGCGGCGGCGGGAATGGCGGGTCTGATTAAAGTCGTTCTCTGCCTACAACA
                  CCAAGAAATTCCCCCTAATCTCCACTTTCAAACCCTTAATCCCTATATTTCCCTAGCTGA
                  CACAGCTTTTGCGATTCCCACTCAGGCTCAACCCTGGCGGACCAAACCCCCTAAGTCTGG
                  TGAAAACGGTGTCGAACGACGTTTAGCAGGACTCAGTTCCTTTGGGTTTGGGGGGACAAA
                  TTCCCATGTGATTCTCAGCGAAGCCCCTGTCACCGTTAAAAACAATCAACAAAATGGGCA
                  GAAGTTGATAGAACGTCCCTGGCATTTGCTGACTTTATCTGCCAAGAATGAAGAAGCCTT
                  AAAAGCCTTAGTCCATTGTTATCAAAAGTATTTAGCTGATCATCATGAAATTCCTCTCGC
                  TGATGTTTGTTTTACGGCCAATAGTCGGCGATCGCACTTTAATCATCGTTTAGGAGTAGT
                  GGCTAGAGATCGCTTAGAAATGTTGCAGAAGTTAGAGAACTTTAGTAACCAAGAAAGGAT
                  GAGAGAACCGAAGAGTATTAACAAAAAGAAAAAACCTAAAATTGTTTTTCTATTTGCCGG
                  TCAAGGTTCTCAATATGTAGGTATGGGTCGTCAACTGTACGAAACCCAACCCATCTTTCG
                  CCAAACCTTGGATCGCTGTGCTGAAATCCTGCGACCCCATTTAGATCAACCCCTCTTAGA
                  AATTCTTTATCCTGCTGACCCAGAAGCCGAAACAGCGAGTTTTTACCTAGAGCAGACTGC
                  CTATACCCAACCCACTTTATTCGCATTCGAGTATGCCCTAGCACAGTTATGGCGTTCCTG
                  GGGAATAGAACCGGCGGCAGTAATTGGTCACAGTGTCGGTGAATATGTGGCGGCCACCGT
                  TGCCGGAGCCTTAAGTCTAGAAGAAGGATTAACGCTAATTGCCAAACGGGCAAAACTGAT
                  GCAGTCTCTCCCCAAGAATGGGACAATGATCGCCGTTTTTGCCGCAGAAGAGCGGGTTAA
                  AGCTGTTATTGAGCCTTATAGGACTGATGTAGCGATCGCTGCTGTTAATGGACCAGAAAA
                  TTTTGTTATTTCAGGAAAAGCGCCGATTATTGCTGAGATTATCATTCATTTAACGGCAGC
                  AGGAATAGAAGTTCGTCCTCTCAAAGTTTCCCATGCTTTTCACTCGCACCTGTTGGAGCC
                  AATTTTAGATTCCTTAGAACAGGAAGCTGCTGCTATTTCCTACCAACCCCTGCAAATTCC
                  CTTAGTTGCTAATTTAACGGGGGAAGTTCTACCAGAAGGAGCAACGATTGAGGCTCGTTA
                  CTGGCGAAATCATGCACGCAACCCTGTACAATTTTATGGGAGTATCCAAACGCTGATCGA
                  GCAGAAATTCAGTCTTTTTTTAGAAGTTAGCCCTAAACCGACTTTATCTCGATTGGGTCA
                  ACAATGTTGTCCAGAAAGATCGACCACTTGGCTATTTTCCCTCGCCCCTCCTCAAGAAGA
                  AGAACAAAGCCTACTAAATAGTTTGGCGATTCTCTATGATTCCCAAGGAGCCGAAATAAA
                  CTGGGAAGGGTTTAATCAAAATTATCCCCACCATTTACTGGCTCTACCGACCTATCCTTT
                  TCAACGTCAACGCTATTGGCTTGAAACCGGTAAACCGACTTCTGAAGAAACAACCATGAC
                  GACCAATGCCACTAATGTCCAAGCTATCTCCAGCCATCAAAAACAACAGGAGATTCTAAT
                  CACATTGCAAACCCTAGTGGGAAATTTACTGCAATTGTCCCCTGCTGATGTCAATGTTCA
                  TACACCTTTCCTGGAGATGGGGGCAGATTCCATTGTCATGGTTGAGGCGGTCAGACGGAT
                  TGAGAATACCTATAACGTTAAAATTGCTATGCGTCAGTTATTTGAGGAGTTATCTACTTT
                  AGATGCTTTAGCTACTTATTTAGCTCAAAATCCGGCTACTGATTGCCAAACTGCTCAAAT
                  TAATACCGAGGTGTTTTCTGCGCCCATTGCCTGCTCAAATAACCGATCGCCCAATGTCGT
                  GCTGAGTTCTAATACCAACGGCTTTCAACGTCAAACAGCTTCTCCAGGTTTTTCGGCGAT
                  CGCCCCCCTTGCAGGAATGGGAGGAGCAGGGGAAATGGGAGGAGTTGAAGTGCCTCAAGT
                  TTCTGTGCCACAAACCAGTGCGGTAACAGCCTCAGGTTCAACCGTTTCTAGTTCTGCCCT
                  GGAAAACATTATGGGTCAACAGTTACAACTGATGGCCAAACAGTTAGAAGTCTTGCAAAC
                  GGCCAATTTTGCCCCGACGACTCCCCGAACCACAGAAAATTCCCCATCTTCCGTCAGTCA
                  AAATAGGTCAAACGGACTTACACAACAGTTAATTCCCCCCCAGCAATTAGCGGCGAACCT
                  AGAGCCAATAGCCAGTCGCACCCGTCAAACCAGCAATCAAGCTTCTGCTCCTAAACCGAC
                  AGTAACAGCCACTCCCTGGGGGCCGAAAAAACCACCCACAGGTGGATTCACTCCCCAACA
                  ACAGCAACATCTAGAGGCATTAATTGCTCGCTTTACGGAACGTACCAAAACCTCTAAGCA
                  AATTGTGCAAAGCGATCGCCTGCGTTTAGCAGATAGTCGAGCCTCGGTCGGATTCCGTAT
                  GTCTATTAAAGAGATGCTTTATCCCATTGTGGCCCAACGTTCTCAAGGATCAAGAATTTG
                  GGATGTGGACGGTAATGAATATATTGATATGACGATGGGGCAAGGGGTAACGCTGTTTGG
                  GCATCAACCAGACTTCATTATGTCGGCCCTACAAAGCCAACTCACTGAAGGCATTCATCT
                  CAATCCGCGATCGCCAATTGTGGGAGAAGTGGCCGCCTTAATTTGTGAACTAACAGGAGC
                  CGAACGAGCTTGTTTTTGCAACTCTGGAACCGAAGCCGTAATGGCCGCTATTCGTATCGC
                  CAGGGCAACAACAGGTCGGAGTAAAATTGCCCTCTTTGAAGGCTCCTATCATGGACATGC
                  GGACGGAACCCTTTTTAGGAACCAAATTATTGATAACCAACTCCACTCTTTTCCCCTAGC
                  TCTAGGCGTTCCCCCCAGCCTTAGTTCCGATGTGGTGGTATTGGACTATGGCAGTGCGGA
                  AGCTCTGAACTATTTACAAACCCAGGGGCAGGATTTAGCGGCGGTCTTAGTAGAACCAAT
                  TCAAAGTGGCAATCCTCTACTCCAACCCCAACAATTTCTCCAAAGTCTGCGACAAATTAC
                  CAGTCAAATGGGCATTGCCCTGATTTTTGATGAAATGATTACGGGTTTTCGATCGCACCC
                  AGGGGGAGCGCAAGCTTTATTTGGAGTACAGGCGGATATTGCCACCTATGGCAAAGTAGT
                  TGCGGGAGGAATGCCCATTGGAGTTATTGCAGGTAAGGCCCATTATCTGGACAGCATTGA
                  CGGGGGAATGTGGCGTTATGGCGATAAATCCTATCCTGGGGTGGACAGAACCTTTTTTGG
                  GGGAACCTTTAATCAGCATCCGTTAGCAATGGTAGCGGCTAGGGCTGTCCTGACCCATTT
                  AAAGGAGCAGGGGCCAGGTCTGCAACAACAATTAACTGAACGCACTGCGGCCTTAGCCGA
                  TACACTGAATCATTATTTTCAAGCCGAAGAAGTTCCTATTAAAATCGAACAGTTTAGTTC
                  TTTCTTCCGGTTTGCCCTCTCTGGCAATTTGGATTTACTTTTCTATCACATGGTAGAAAA
                  AGGTATTTATGTCTGGGAATGGCGTAAACATTTTCTTTCAACCGCCCATACGGAAGCCGA
                  TCTTGCCCAATTTGTCCAAGCGGTTAAGGATAGCATCACAGAATTGCGTCAGGGAGGTTT
                  TATCCCCGCAAAAAAGCCTTCCTGGCCAGTGCCAACGCCTCAAATTGATCCCCCCCTAAC
                  CCCCCTTGATAAGGGGATTGATCCCCCCCTAACCCCCCTTGATAAGGGGATTGATCCCCC
                  CCTAACCCCCCTTGATAAGGGGGGAGATGTTGATGTCGCGCTTGATAAGGGAGGAAATTC
                  TCATTCTGTTAGGGACAGTAAGTTAGGGAAAGGGAGCGGGTCTCAAGACCAAAAAACGAT
                  ACAGTTTAGCCTCTACTACTTTGGTAGCTATGAAGCGGAATTTAACCCGAATAAATATAA
                  CTTACTGTTTGAAGGAGCTAAATTTGGCGATCGCGCTGGTTTTACGGCCCTTTGGATTCC
                  TGAACGTCATTTCCACGCTTTTGGTGGTTTTTCTCCCAATCCTTCGGTTTTGGCGGCGGC
                  TTTAGCACGGGAAACCAAACAGATTCAACTGCGATCAGGCAGTGTGGTTTTACCGCTACA
                  TAATTCCATCCGAGTCGCCGAAGAATGGGCAGTGGTGGACAATCTTTCCCAGGGCCGCGT
                  TGGTATTGCTTTTGCATCGGGTTGGCATCCCCAGGATTTTGTCTTGGCTCCCCAGTCCTT
                  TGGCCAACATCGGGAATTGATGTTCCAAGAAATTGAAACCGTCCAGAAACTTTGGCGAGG
                  GGAAGCGATCACCGTGCCAGACGGAAAGGGTCAAAGGGTAGAGGTTAAAACCTATCCCCA
                  ACCGATGCAGTCCCAGTTACCCAGCTGGATTACTATTGTCAATAATCCCGATACCTATAT
                  CAGAGCAGGGGCGATCGGTGCTAATATCCTTACCAATCTGATGGGGCAAAGCGTGGAAGA
                  TTTAGCCCGTAATATTGCGCTATATCGTCAATCTTTGGCAGAGCATGGTTATGATCCCGC
                  GTCGGGAACGGTGACAGTTCTCCTGCATACTTTTGTTGGCAAGGATTTAGAACAAGTTCG
                  AGAACAGGCTCGCCAACCCTTTGGGCAATACCTCACCTCCTCTGTCGGACTCTTGCAGAA
                  CATGGTCAAGAGCCAGGGCATGAAAGTGGATTTTGAACAATTAAGAGACGAAGATCGGGA
                  CTTTCTCCTCGCTTCTGCCTATAAACGCTATACAGAAACCAGTGCTTTAATTGGCACACC
                  CGAATCCTGTCGTCAAATTATTGATCATTTGCAGTCCATCGGTGTGGATGAAGTGGCTTG
                  TTTTATTGATTTTGGGGTAGATGAACAAACAGTTTTGGCCAATTTACCCTATCTCCAGTC
                  CCTAAAAGACTTATATCAACCTCATCTCCCCCCTTATCAAGGGGGGTTAGGGGGGGATCA
                  ATCCCCTTATCAAGGGGGGTTAGGGGGGGATCAATCCCCTTATCAAGGGGGGTTAGGGGG
                  TGATCAATCCCCTTATCAAGGGGGGTTAGGGGGTGATCAATCCCCTTATCAAGGGGGGTT
                  AGGGGGGGATCAATCCCCTTATCAAGGAGAGTTAGGGGGGGATCAATCCCCTTATCAAGG
                  GGGGTTAGGGGGGGATCAAGTCCCTCTCACCGAAGCCCAACGACAACTGTGGATTTTGGC
                  TCAATTAGGAGACAACGGCTCTGTGGCCTATAACCAATCAGTGACATTGCAATTAAGTGG
                  CCCATTAAATCCCGTCGCAATGAATCAAGCTATTCAACAAATCAGCGATCGCCATGAAGC
                  GTTACGAACCAAAATTAATGCCCAGGGAGATAGTCAAGAAATCCTGCCCCAGGTCGAAAT
                  TAACTGCCCTATCTTAGACTTCAGTCTTGACCAAGCTTCGGCCCAACAGCAAGCAGAACA
                  ATGGTTAAAGGAAGAAAGTGAAAAACCCTTTGATTTGAGCCAGGGTTCTCTCGTGCGTTG
                  GCATCTACTCAAATTAGAACCAGAATTACATTTGTTAGTATTAACGGCCCATCACATTAT
                  CAGTGACGGTTGGTCAATGGGGGTAATCCTTCGGGAATTAGGAGAGTTATATTCAGCCAA
                  ATGTCAGGGTGTTACGGCTAATCTTAAAACCCCAAAACAGTTTCGAGAATTGATTGAATG
                  GCAAAGCCAGCCAAGCCAAGGGGAAGAACTGAAAAAACAGCAAGCCTATTGGTTAGCAAC
                  CCTTGCCGATCCCCCTGTTTTGAATTTACCCACTGACAAACCTCGTCCAGCTTTACCCAG
                  TTACCAAGCTAATCGTCGAAGTCTAACTTTAGATAGCCAATTTACAGAAAAACTAAAGCA
                  ATTTAGTCGTAAACAGGGCTGTACCTTGCTGATGACCCTGTTATCGGTTTATAACATTCT
                  CGTTCATCGTTTGACGGGACAGGATGATATTCTGGTGGGTCTGCCAGCCTCTGGACGGGG
                  GCTTTTAGATAGTGAAGGTATGGTGGGTTATTGCACCCATTTTTTACCAATTCGCAGTCA
                  ATTAGCAGGTAATCCCACTTTTGCTGAATATCTCAAACAAATGCGGGGGGTTTTGTTGTC
                  GGCTTATGAACATCAGGACTATCCCTTTGCTCTTTTGCTCAATCAGTTAGATTTACCGCG
                  TAATACCAGTCGCTCTCCTTTAATTGATGTCAGTTTCAATTTAGAACCAGTTATTAACCT
                  ACCCAAAATGAAAGGATTAGAGATTAGTTTGTTGCCTCAAAGTGTAAGTTTTAAGGATCG
                  AGATTTGCATTGGAATGTGACAGAAATGGGTGGAGAAGCTCTGATTGATTGTGACTACAA
                  TACAGACTTATTTAAAGATGAAACGATTCAGCGTTGGTTAGGCCATTTTCAAACCTTACT
                  TGAGGCAGTTATTAATGATTCGCAACAAAATCTGCGGGAATTACCCTTATTAAGTTCTGC
                  TGAACGACAACAGTTATTAGTGGATTGGAATCAAACCAAGACCGACTATCCCCAAGATCA
                  GTGTATTCATCAATTATTTGAAGCGCAAGTTGAACGGACTCCCGATGCGATTGCGGTGGT
                  ATTTGAAACTCAACAATTAACTTACAGTGAATTAAATTGTCGAGCCAATCAGTTAGCACA
                  TTATTTACAAAAATTAGGAGTTGGGCCAGAGGTCTTAGTCGGTATTTTGGTCGAACGTTC
                  TTTAGAAATGATTGTCGGATTGTTAGGGATTCTCAAGGCTGGGGGAGCCTATGTACCTCT
                  TGATCCTGACTATCCCCCTGAACGTCTTCAATTTATGTTAGAAGATAGTCAATTTTTTCT
                  CCTCTTAACCCAACAGCATTTACTGGAATCTTTTGCTCAGTCTTCAGAAACGGCTACTCC
                  CAAGATTATTTGTTTGGATAGCGACTACCAAATTATTTCCCAGGCAAAGAATATTAATCC
                  CGAAAATTCAGTCACAACGAGTAATCTTGCCTATGTAATTTATACCTCTGGTTCGACAGG
                  TAAACCGAAGGGCGTGATGAATAATCATGTTGCTATTAGTAATAAATTGTTATGGGTACA
                  AGACACTTATCCTCTAACCACAGAAGACTGTATTTTACAAAAAACTCCCTTTAGTTTTGA
                  TGTTTCAGTGTGGGAATTATTCTGGCCCCTACTAAACGGAGCGCGTTTGGTTTTTGCCAA
                  GCCGAATGGCCATAAAGATGCCAGTTACTTAGTCAATCTGATTCAAGAGCAACAAGTAAC
                  AACGCTACATTTTGTGTCTTCTATGCTACAGCTTTTTCTGACAGAAAAAGACGTAGAAAA
                  ATGTAATAGTCTTAAACGAGTCATTTGTAGTGGTGAAGCCCTTTCTTTAGAGCTTCAAGA
                  ACGTTTTTTTGCTCGTTTAGTCTGTGAATTACACAATCTTTATGGACCGACAGAAGCCGC
                  TATTCATGTCACATTTTGGCAATGTCAATCAGATAGCAATTTGAAAACAGTACCCATTGG
                  TCGGCCGATCGCTAATATCCAAATTTACATTTTAGACTCTCATCTTCAGCCAGTACCTAT
                  TGGAGTAATCGGAGAATTGCACATTGGTGGGGTTGGTTTGGCGCGGGGTTATTTAAACAG
                  GCCTGAGTTAACGGCGGAGAAATTTATTGCAAATCCGTTTGCTTCCCTTGATCCCCCCCT
                  AACCCCCCTTGATAAGGGGGGAGATGAGAGCTATAAAACTTTTAAAAAGGGGGGAGAGCA
                  ACCATCAAGATTGTATAAAACGGGAGATTTAGCTCGTTATTTACCCGATGGCAAGATTGA
                  GTATCTAGGGCGCATTGATAATCAGGTAAAAATTCGCGGTTTCCGGATTGAATTGGGGGA
                  AATTGAAGCGGTTTTGCTATCCCATCCCCAGGTACGAGAAGCGGTCGTTTTGGTGAGCGA
                  AAGCGATCGCTCTGAAAATCGGGCTTTGGTCGCTTATATTGTCCCTAATGATCCTGCTTG
                  TACGACTCAATCATTACGAGAGTTTGTTAAACGGCAGCTTCCTGACTATATGATCCCAGC
                  TTATTGGCTGATCCTTGACAATTTACCGTTAACCAGCAATGGCAAAATTGATCGTCGGGC
                  TTTACCGTTACCTAATCCAGAGTTAAATCGTTCGATAGACTATGTGGCTCCCAAAAATCC
                  TACCCAGGAGGCGATCGCCGCTATTTTTGGTCAAGTTTTAAAACTGGAAAAAGTGGGAAT
                  TTATGATAACTTTTTTGAGATCGGCGGTAATTCTTTGCAAGCCACTCAAGTTATTTCACG
                  CTTACGAGAAAGTTTTGCCCTAGAGTTGCCCTTGCGTCGCCTGTTTGAACAACCGACTGT
                  GGCGGATTTGGCTTTAGCCGTAACGGACATTCATGCCACTTTACAAAAATTACAAACCCC
                  TATTGATGATTTATCAGGCGATCGCGAGGAGATTGAACTATGAAATCTATTGAAACCTTT
                  TTGTCAGATTTAGCCAATCAAGATATTAAACTCTGGATGGACGGCGATCGCCTGCGTTGT
                  AATGCACCCCAGGGCCTATTAACCCCAGAGATTCAAACAGAACTGAAAAACCGTAAAGCA
                  GAAATCATTCACTTTCTCAATCAACTGGGTTCAGAGGAGCAAATTAATCCTAGAACGATT
                  CTTCCCATTCCTCGTGATGGCCAATTACCCCTCTCCTTTGCCCAGTCGCGACTCTGGTTC
                  TTGTATCAATTAGAAGGAGCCACGGGAACCTATAACATGACAGGGGCCTTGAGTTTAAGC
                  GGGCCTCTTCAGGTCGAAGCCCTCAAACAAGCCCTAAGAACTATCATTCAACGCCATGAG
                  CCATTGCGTACCAGTTTCCAATCGGTTGACGGGGTTCCAGTGCAGGTGATTAATCCCTAT
                  CCTGTTTGGGAATTAGCGATGGTTGATTTGACAGGAAAGGAGACAGAAGCAGAAAAATTG
                  GCCTATCAGGAATCCCAAACCCCGTTTGATTTGACCAATAGTCCTTTGTTGAGGGTAACG
                  CTCCTCAAATTACAGCCAGAAAAGCATATTTTATTAATTAATATGCACCATATTATTTCC
                  GATGGCTGGTCAATCGGTGTTTTTGTTCGTGAATTGTCCCATCTCTATAGGGCTTTTGTG
                  GCGGGTAAAGAACCAACTTTACCGATTTTACCAATTCAGTATGCGGATTTTGCCGTTTGG
                  CAGCGAGAGTGGTTACAGGGTAAGGTTTTAGCGGCTCAATTGGAATATTGGAAGCGACAA
                  TTGGCAGATGCTCCTCCTCTGCTGGAACTGCCCACTGATCGCCCTCGTCCCGCAATCCAA
                  ACCTTTCAAGGCAAGACAGAAAGATTTGAGCTAGATAGGAAACTGACCCAAGAATTAAAG
                  GCATTAAGTCAACAGTCGGGTTGTACTTTATTTATGACTTTGTTGGCCGCTTTTGGGGTG
                  GTTTTATCCCGTTATAGTGGCCAGACTGATATCGTCATTGGTTCGGCGATCGCCAACCGT
                  AATCGCCAAGACATTGAGGGGTTAATTGGCTTTTTTGTTAACACTTTGGCGTTGAGGTTA
                  GATTTATCAGAAAAACCCAGCTTTGCCGCTTTTTTAAAACAAGTACAGGAAGTCACTCAG
                  GATGCCTATGAGCATCAAGACTTGCCCTTTGAAATGTTAGTGGAAGAATTACAACTAGAG
                  CGCAAATTAGACCGAAATCCTTTGGTACAGGTGATGTTTGCCCTACAAAATGCGGCCAAT
                  GAAACCTGGAATTTACCTGGGTTGACCATTGAAGAAATGTCTTGGGAACTTGAACCTGCC
                  CGTTTTGACCTAGAGGTTCATTTATCAGAAGTTAACGCCGGCATAGCTGGATTCTGTTGC
                  TACACCATTGATCTATTTGATGATGCAACGATCGCCCGTCTATTGGAACATTTTCAGAAT
                  CTTCTCAGGGCAATTATTGTTAATCCTCAAGAATCGGTAAGTTTATTACCCTTGTTGTCA
                  GAACAGGAAGAAAAGCAACTTTTAGTTGATTGGAATCAAACCCAAGCCGATTATCCCCAA
                  GATAAGCTTGTCCATCAGTTATTTGAAGTTCAAGCAGCCAGTCAGCCAGAAGCGATCGCT
                  CTAATCTTTGAAAATCAGGTTTTGACCTATGGAGAATTAAACCATCGCGCCAATCAATTA
                  GCTCACTATCTTCAGTCGTTAGGAGTCACCAAAGAACAAATCGTCGGGGTTTATCTGGAA
                  CGTTCCCTTGAAATGGCGATCGGATTTTTAGGTATTCTCAAAGCAGGAGCCGCCTATCTC
                  CCCATTGATCCTGAATATCCCTCAGTACGCACCCAATTTATTCTCGAAGATACCCAACTT
                  TCGCTTCTCTTAACTCAGGCAGAACTGGCAGAAAAACTGCCCCAGACTCAAAACAAAATT
                  ATCTGTCTAGATCGGGACTGGCCAGAAATTACCTCCCAACCCCAGACAAACCTAGACCTA
                  AAGATAGAACCTAATAACCTAGCCTATTGCATCTATACTTCTGGTTCCACAGGACAACCC
                  AAAGGAGTACTGATTTCCCATCAAGCCCTACTCAACTTAATTTTCTGGCATCAACAAGCG
                  TTTGAGATTGGCCCCTTACATAAAGCGACCCAAGTGGCAGGCATTGCTTTCGATGCAACG
                  GTTTGGGAATTGTGGCCCTATCTGACCACAGGAGCCTGTATTAATCTGGTTCCCCAAAAT
                  ATTCTGCTCTCACCGACGGATTTACGGGATTGGTTGCTTAACCGAGAAATTACCATGAGT
                  TTTGTGCCAACTCCTTTAGCTGAAAAATTATTATCCTTGGATTGGCCTAACCATTCTTGT
                  CTAAAAACCCTGTTACTGGGAGGTGACAAACTTCATTTTTATCCTGCTGCGTCCCTTCCC
                  TTTCAGGTCATTAACAACTATGGCCCAACGGAAAATACAGTGGTTGCGACCTCTGGACTG
                  GTCAAATCATCTTCATCTCATCACTTTGGAACTCCGACTATTGGTCGTCCCATTGCCAAC
                  GTCCAAATCTATTTATTAGACCAAAACCTACAACCTGTCCCCATTGGTGTACCAGGAGAA
                  TTACATTTAGGTGGGGCGGGTTTAGCGCAGGGCTATCTCAATCGTCCTGAGTTAACGGCT
                  GAAAAATTTATTGCCAATCCCTTTGATCCCCCCCTAACCCCCCTTGATAAGGGGGGAGAA
                  GAACCCTCAAAACTCTATAAAACGGGAGACTTAGCCCGTTATTTACCCGATGGCAATGTA
                  GAATTTTTGGGACGTATTGACAATCAGGTAAAAATTCGGGGTTTTCGCATCGAAACTGGG
                  GAAATCGAAGCCGTTTTAAGTCAATATTTCCTATTAGCTGAAAGTGTAGTCGTTGCCAAG
                  GAAGATAATACTGGGGATAAACGCCTCGTGGCTTATTTGGTTCCCGCCTTGCAAAATGAG
                  GCCCTACCAGAGCAATTAGCCCAATGGCAAAGTGAATACATCAGTGATTGGCAAAGTCTC
                  TATGAAAGAACCTATAGTCAAGGGCAAGACAGCCTAGCTGATCTCACTTTTAATATCACG
                  GGTTGGAATAGCAGTTATACTCGTCAACCCCTTCCTGCTTCAGAAATGCGAGAGTGGGTC
                  GAAAACACTGTTAGTCGCATCTTGGCTTTCCAACCAGAACGCGGTTTAGAAATTGGTTGT
                  GGTACAGGTTTGTTACTCTCCAGGGTAGCAAAGCATTGTCTTGAATATTGGGCAACGGAT
                  TATTCCCAAGGGGCGATCCAGTATGTTGAACGGGTTTGCAATGCCGTTGAAGGTTTAGAA
                  CAGGTTAAATTACGCTGTCAAATGGCAGATAATTTTGAAGGTATTGCCCTACATCAATTT
                  GATACCGTCGTCTTAAATTCGATTATTCAGTATTTTCCCAGTGTGGATTATCTGTTACAG
                  GTGCTTGAAGGGGCGATCAACGTCATTGGCGAGCGAGGTCAGATTTTTGTCGGGGATGTG
                  CGGAGTTTACCCCTATTAGAGCCATATCATGCGGCTGTGCAATTAGCCCAAGCTTCTGAC
                  TCGAAAACTGTTGAACAATGGCAACAACAGGTGCGTCAAAGTGTAGCAGGTGAAGAAGAA
                  CTGGTCATTGATCCCACATTGTTCCTGGCTTTAAAACAACATTTTCCGCAAATTAGCTGG
                  GTAGAAATTCAACCGAAACGGGGTGTGGCTCACAATGAGTTAACTCAATTTCGCTATGAT
                  GTCACTCTCCATTTAGAGACTATCAATAATCAAGCATTATTGAGCGGCAATCCAACGGTA
                  ATTACCTGGTTAAATTGGCAACTTGACCAACTGTCTTTAACACAAATTAAAGATAAATTA
                  TTAACAGACAAACCTGAATTGTGGGGAATTCGTGGTATTCCTAATCAGCGAGTTGAAGAG
                  GCTCTAAAAATTTGGGAATGGGTGGAAAATGCCCCTGATGTTGAAACGGTTGAACAACTC
                  AAAAAACTTCTCAAACAACAAGTAGATACTGGTATTAATCCTGAACAGGTTTGGCAATTA
                  GCTGAGTCTCTCGGTTACACCGCTCACCTTAGTTGGTGGGAAAGTAGTCAAGACGGTTCC
                  TTTGATGTCATTTTTCAGCGGAATTCAGAAGCGGAGGACTCAAAAAAATTAACCCTTTCA
                  AAACTTGCTTTCTGGGATGAAAAACCCTTTAAAATAAAGCCCTGGAGTGACTATACTAAC
                  AACCCTCTGCGCGGTAAGTTAGTCCAAAAATTAATTCCTAAAGTACGAGAATTTCTGCAA
                  GAAAAACTACCCAGTTATATGGTTCCCCAGGCGTTTGTGCTGCTTGATTCCCTTCCTTTG
                  ACCCCCAATGGTAAGGTGGATCGTAAGGCGTTACCTTCTCCTGATGCGGCGACTCGTGAT
                  TTAGCGAACAGTTTTGTCTTACCCCGCAATCCGATTGAAGCTCAACTGACTCAAATTTGG
                  AGTGAAGTTTTGGGACTGGAACGCATTGGCGTTAAGGACAACTTTTTTGAATTGGGAGGA
                  CATTCTCTTTTGGCTACCCAGGTTTTATCAAGAATTAATTCAGCCTTTGGACTTGATCTT
                  TCTGTGCAAATTATGTTTGAATCACCAACGATCGCGGGCATTGCGGGTTATATTCAAGCG
                  GTAGATTGGGTCGCCCAGGATCAAGCCGATAGCTCGTTAAATCATGAAAATACTGAGGTA
                  GTGGAGTTCTAAGTTATGACGAAAAAGATTGTTGAATTTGTCTGTTATCTACGGGATTTA
                  GGCATTACTTTAGAAGCTGATGAAAACCGCTTACGCTGTCAGGCTCCCGAAGGAATTTTG
                  ACCCCAGCACTCCGTCAAGAAATTGGCGATCACAAACTGGAATTATTACAATTTTTACAA
                  TGGGTCAAACAGTCTAAAAGTACCGCTCATTTGCCTATTAAACCTGTCGCTAGAGACGGT
                  CATTTACCCCTGTCTTTTGCTCAACAACGTTTATGGTTTTTACATTATCTTTCCCCTGAT
                  AGTCGTTCCTACAATACCCTGGAAATATTGCAAATTGATGGGAATCTCAATCTGACTGTG
                  CTAGAGCAGAGTTTGGGGGAATTAATTAACCGCCATGAAATTTTTAGAACAACATTCCCC
                  ACTGTTTCAGGGGAACCGATTCAGAAAATTGCACTTCCTAGTCGTTTTCAGTTAAAAGTT
                  GATAATTATCAAGATTTAGACGAAAATGAACAATCAGCTAAAATTCAACAAGTAGCAGAA
                  TTGGAAGCAGGACAAGCTTTTGATTTAACGGTGGGGCCACTGATTCAGTTTAAGCTATTG
                  CAATTGAGTCCCCAGAAGTCGGTGCTGCTGTTGAAAATGCACCATATTATCTATGATGGC
                  TGGTCTTTTGGGATTCTGATTCGGGAATTATCGGCTCTATACGAAGCATTTTTAAAGAAC
                  TTAGCCAATCCTCTCCCTGCGTTGTCTATTCAGTATGCAGATTTTGCGGTTTGGCAACGT
                  CAATATCTCTCAGGTGAGGTCTTAGATAAACAACTCAATTATTGGCAAGAACAGTTAGCA
                  ACAGTCTCTCCTGTTCTTACTTTACCAACGGATAGACCCCGTCCGGCGATACAAACTTTT
                  CAGGGAGGAGTTGAGCGTTTTCAACTGGATCAAAATGTCACTCAAGGTCTTAAAAAGTTA
                  GGTCAAGATCAGGTTGCAACCCTGTTTATGACGTTGTTGGCCGGTTTCGGCGTTTTGCTA
                  TCTCGTTATAGTGGTCAATCTGATCTGATGGTGGGTTCTCCGATCGCTAATCGTAATCAA
                  GCAGCGATCGAACCTTTAATTGGCTTTTTTGCTAACACTTTGGCTTTAAGAATTAATTTA
                  TCAGAAAATCCCAGTTTTTTAGAATTATTAGAACAAGTTAAACAGACAACTTTAGAGGGT
                  TATGCTCACCAAGACCTACCCTTTGAGATGTTAGTAGAAAAGCTACAACTTGACCGTGAT
                  TTGAGCAGAAATCCTTTAGTACAAGTCATGTTTGCGCTACAAAATACCTCTCAAGATACT
                  TGGAATCTTTCGGGTTTAAGTATTGAAAGTTTATCTTTATCAGTGGAAGAAACTGTCAGA
                  TTTGATCTAGAAGTAAACTGCTGGCAAAATTCAGAAGGTTTAGCAATAGATTGGATTTAC
                  AGCAGAGATTTATTTGACACTGCAACAATTGCAAGAATGGGAGAACATTTTCAAAATTTA
                  GTTCAGGCAATCATACTCAATCCAAAAGCTACAGTTAAAGAACTTCCTTTATTAACACCC
                  AAGGAACGTGAGCAATTATTAATATCTTGGAATAATAGCAAGACTGATTATCCTCAAGAG
                  CAGTGTATTTATCAATTATTTGAAGCACAAGTTGAACGGACTCCAAAGGCGATCGCAGTG
                  GTATTTGAGGAGCAATCATTAACATACACTGAATTAAACCATCGCGCTAATCAGTTAGCC
                  CATTATTTACAAACTTTAGGCGTGGGAGCAGAAGTCTTAGTCGGTATTTCCCTAGAACGT
                  TCTTTAGAGATGATTATCGGCTTATTAGGGATTCTCAAGGTAGGTGGTGCTTATCTTCCT
                  CTTGATCCAGACTATCCCACTGAGCGTCTTCAGTTGATGTTAGAAGACAGTCAAGTTCCT
                  TTTTTGATTACCCACAGTTCTTTATTAGCAAAATTGCCTCCCTCTCAAGCAACTCTGATT
                  TGTTTAGATCATATCCAAGAGCAGATTTCTCAATATTCTCCAGATAATCTTCAATGTCAG
                  TTAACTCCTGCCAATTTAGCTAACGTTATTTATACCTCTGGCTCTACGGGTAAGCCTAAA
                  GGGGTGATGGTTGAACATAAAGGTTTAGTTAACTTAGCTCTTGCTCAAATTCAATCTTTT
                  GCAGTCAACCATAACAGTCGTGTGCTGCAATTTGCTTCTTTTAGTTTTGATGCTTGTATT
                  TCAGAAATTTTGATGACCTTTGGTTCTGGAGCGACGCTTTATCTTGCACAAAAAGATGCT
                  TTATTGCCAGGTCAGCCATTAATTGAACGGTTAGTAAAGAATGGAATTACTCATGTGACT
                  TTGCCGCCTTCAGCTTTAGTGGTTTTACCCCAGGAACCGTTACGCAACTTAGAAACCTTA
                  ATTGTGGCGGGTGAGGCTTGTTCTCTTGATTTAGTGAAACAATGGTCAATCGATAGAAAC
                  TTTTTCAATGCCTATGGGCCAACGGAAGCGAGTGTTTGTGCCACTATTGGACAATGTTAT
                  CAAGATGATTTAAAGGTGACGATTGGTAAGGCGATCGCCAATGTCCAAATTTATATTTTA
                  GATGCCTTTTTACAGCCGGTGCCGGTGGGAGTGTCAGGAGAGTTATACATTGGTGGAGTT
                  GGGGTGGCAAGGGGCTATTTAAATCGTCCTGAATTAACCCAAGAAAAATTTATTGCTAAT
                  CCTTTTAGTAACGACCCAGATTCTCGGCTCTATAAAACTGGCGACTTAGCGCGTTATTTA
                  CCCGATGGTAATATTGAATATTTAGGACGCATTGACAATCAGGTAAAAATTCGCGGTTTT
                  CGCATTGAGTTAGGAGAAATTGAAGCGGTTCTGAGTCAATGTCCCGATGTGCAAAATACG
                  GCGGTGATTGTCCGCGAAGATACTCCTGGCGATAAGCGCTTAGTTGCCTATGTGGTTCTT
                  ACTTCTGACTCCCAGATAACTACTAGCGAACTGCGTCAATTTTTGGCGAATCAATTACCC
                  GCCTATCTTGTTCCTAATACCTTTGTTATTTTAGATGATTTGCCCCTAACCCCCAGTGGC
                  AAATGCGATCGCCGTTCCTTACCTATACCCGAAACACAAGCGTTATCAAATGACTATATT
                  GCCCCTAAATCTCCCACTGAAGAAATTCTGGCTCAAATATGGGGGCAAGTTCTCAAGATA
                  GAAAGAGTCAGCAGAGAAGATAATTTCTTTGAATTGGGGGGGCATTCCCTTTTAGCTACC
                  CAGGTAATGTCCCGTCTGCGTGAAACTTTTCAAGTCGAATTACCTTTGCGTAGTCTCTTT
                  ACCGCTCCCACTATTGCTGAATTGGCCCTAACAATTGAGCAATCTCAGCAAACCATTGCT
                  GCTCCCCCCATCCTAACCAGAAACGACAGTGCTAACCTCCCGTTATCTTTTGCTCAACAA
                  CGTTTATGGTTTCTGGATCAATTAGAACCTAACAGCGCCTTTTATCATGTAGGGGGAGCC
                  GTAAGACTAGAAGGAACATTAAATATTACTGCCTTAGAGCAAAGCTTAAAAGAAATTATT
                  AATCGTCATGAAGCTTTACGCACAAATTTTATAACGATTGATGGTCAAGCCACTCAAATT
                  ATTCACCCTACTATTAATTGGCGATTGTCTGTTGTTGATTGTCAAAATTTAACCGACACT
                  CAATCTCTGGAAATTGCGGAAGCTGAAAAGCCCTTTAATCTTGCTCAAGATTGCTTATTT
                  CGTGCTACTTTATTCGTGCGATCACCGCTAGAATATCATCTACTCGTGACCATGCACCAT
                  ATTGTTAGCGATGGCTGGTCAATTGGAGTATTTTTTCAAGAACTAACTCATCTTTACGCT
                  GTCTATAATCAGGGTTTACCCTCATCTTTAACGCCTATTAAAATACAATATGCTGATTTT
                  GCGGTCTGGCAACGGAATTGGTTACAAGGTGAAATTTTAAGTAATCAATTGAATTATTGG
                  CGCGAACAATTAGCAAATGCTCCTGCTTTTTTACCTTTACCGACAGATAGACCTAGGCCC
                  GCAATCCAAACTTTTATTGGTTCTCATCAAGAATTTAAACTTTCTCAGCCATTAAGCCAA
                  AAATTGAATCAACTAAGTCAGAAGCATGGAGTGACTTTATTTATGACTCTCCTGGCTGCT
                  TTTGCTACCTTACTTTACCGTTATACAGGACAAGCAGATATTTTAGTTGGTTCTCCTATT
                  GCTAACCGTAATCGTAAGGAAATTGAGGGATTAATCGGCTTTTTTGTTAATACATTAGTT
                  CTGAGATTGAGTTTAGATAATGATTTAAGTTTTCAAAATTTGCTAAACCATGTTAGAGAG
                  GTTTCTTTAGCAGCCTACGCCCATCAAGATTTACCTTTTGAAATGTTAGTAGAAGCACTA
                  CACCCTCAACGAGATCTCAGTCATACCCCTTTATTTCAGGTAATGTTTGTTTTGCAAAAT
                  ACACCAGTGGCTGATCTAGAACTTAAAAATGTAAAGGTTTGTCCTCTACCGATGGAAAAT
                  AAGACTGCTAAATTTGATTTAACCTTATCAATGGAGAATCTAGAGGAAGGATTGATTGGG
                  GTTTGGGAATATAACACCGATCTATTTAATGGCTCAACCATTGAGCGAATGAGTGGACAT
                  TTTGTCACTTTGTTAGAAGATATTGTTGCCGCTCCAACGAAGTCAGTTTTACGGTTGTCT
                  TTGCTGACGCAAGAGGAAAAACTGCAATTATTGATTAAAAATCAGGGTGTTCAAGTTGAT
                  TATTCTCAAGAGCAGTGCATCCATCAATTATTTGAAGCGCAAGTTGAACGGACTCCCGAT
                  GCGATTGCGGTGGTATTTGAGGAGCAATCATTAACCTATGCTGAATTAAATCATCAAGCT
                  AATCAGTTAGTCCATTACTTACAAACTTTAGGAATTGGGCCAGAGGTCTTAGTCGCTATT
                  TCAGTAGAACGTTCTTTAGAAATGATTATCGGCTTATTAGCCATTCTCAAGGCGTGTGGT
                  GCTTATCTCCCTCTTGCTCCTGACTATCCCACTGAGCGTCTTCAGTTCATGTTAGAAGAT
                  AGTCAAGCTTCTTTTTTGATTACCCACAGTTCTTTATTAGAAAAATTGCCTTCTTCTCAA
                  GCGACTCTAATTTGTTTAGATCACATCCAAGAGCAGATTTCTCAATATTCTCCCGATAAT
                  CTTCAAAGTGAGTTAACTCCTTCCAATTTGGCTAACGTTATTTACACCTCTGGCTCTACG
                  GGTAAGCCTAAAGGGGTGATGGTTGAACATCGGGGCTTAGTTAACTTAGCGAGTTCTCAA
                  ATTCAATCTTTTGCAGTCAAAAATAACAGTCGTGTACTGCAATTTGCTTCCTTTAGTTTT
                  GATGCTTGTATTTCAGAAATTTTGATGACCTTTGGTTCTGGAGCGACTCTTTATCTTGCT
                  CAAAAAAATGATTTATTGCCAGGTCAGCCATTAATGGAAAGGTTAGAAAAGAATAAAATT
                  ACCCATGTTACTTTACCCCCTTCAGCTTTAGCTGTTTTACCAAAAAAACCGTTACCCAAC
                  TTACAAACTTTAATTGTGGCGGGTGAGGCTTGTCCTCTGGATTTAGTCAAACAATGGTCA
                  GTCGGTAGAAACTTTTTCAATGCCTATGGCCCGACAGAAACGAGTGTTTGTGCCACGATT
                  GGACAATGTTATCAAGATGATTTAAAGGTCACGATTGGTAAGGCGATCGCTAATGTCCAA
                  ATTTATATTTTGGATGCCTTTTTACAACCAGTACCCATCGGAGTACCAGGGGAATTATAC
                  ATTGGTGGAGTCGGAGTTGCGAGGGGTTATCTAAATCGTCCTGAATTAACGGCGGAAAGA
                  TTTATTCCTAATCCTTTTGATCCCCCCCTAACCCCCCTTAAAAAGGGGGGAGATAAGAGC
                  TATGAAACTTTTAAAAAGGGGGAAGAGCAACCATCAAAACTCTATAAAACGGGAGATTTA
                  GCTCGTTATTTACCCGATGGCAATATTGAATATTTAGGACGCATTGACAATCAGGTAAAA
                  ATTCGCGGTTTTCGCATTGAGTTAGGAGAAATTGAAGCGGTTCTGAGTCAATGTCCCGAT
                  GTGCAAAATACGGCGGTGATTGTCCGTGAAGATACTCCTGGCGATAAACGTTTAGTTGCC
                  TATGTGGTTCTTACTTCTGACTCCCAGATAACTACTAGCGAACTGCGTCAATTCTTGGCT
                  AATCAATTACCTGCCTATCTCGTTCCCAATACCTTTGTTATTTTAGATGATTTGCCCCTA
                  ACCCCCAATGGTAAATGCGATCGCCGTTCCTTACCGCTTCCTGATGATCAGACCAGAAAA
                  AATATTCCTAAAATTGGCCCGCGTAATTTAGTGGAATTACAATTAGCTCAAATCTGGTCA
                  GAGATTTTAGGCATTAATAATATTGGTATTCAGGAAAACTTCTTTGAATTAGGCGGTCAT
                  TCTTTATTAGCAGTCAGTCTGATCAATCGTATTGAACAAAAGTTAGATAAACGTTTACCA
                  TTAACCAGTCTTTTTCAAAATGGAACCATAGCAAGTCTAGCTCAATTACTAGCGCAAGAA
                  ACAACTCAGCCAGCCTCTTCACCGTTGATTGCTATCCAGTCTCAAGGTGATAAAACTCCA
                  TTTTTTGCTGTTCATCCCATTGGTGGTAATGTGCTATGTTATGCCGATTTAGCTCGTAAT
                  TTAGGAACGAAACAGCCGTTTTATGGATTACAATCATTAGGGCTAAGTGAATTAGAAAAA
                  ACTGTAGCCTCTATTGAAGAAATGGCGATGATTTATATTGAAGCAATACAAACTGTTCAA
                  GCCTCTGGTCCCTACTATTTAGGAGGTTGGTCAATGGGAGGAGTGATAGCTTTTGAAATC
                  GCCCAACAATTATTGACCCAAGGTCAAGAAGTTGCTTTACTGGCTTTAATAGATAGTTAT
                  TCTCCCAGTTTACTTAATTCAGTTAATAGGGAGAAAAATTCTGCTAATTCCCTGACAGAA
                  GAATTTAATGAAGATATCAATATTGCCTATTCTTTCATCAGAGACTTAGCAAGTATATTT
                  AATCAAGAAATCTCTTTCTCTGGGAGTGAACTTGCTCATTTTACATCAGACGAATTACTA
                  GACAAGTTTATTACTTGGAGTCAAGAGACGAATCTTTTGCCGTCAGATTTTGGGAAGCAG
                  CAGGTTAAAACCTGGTTTAAAGTTTTCCAGATTAATCACCAAGCTTTGAGCAGCTATTCT
                  CCCAAGACGTATCTGGGTAGAAGTGTTTTCTTAGGAGCGGAAGACAGTTCTATTAAAAAT
                  CCTGGTTGGCATCAAGTAATCAATGACTTGCAATCTCAATGGATTAGCGGCGATCACTAC
                  GGTTTAATTAAAAATCCAGTCCTCGCTGAAAAACTCAATAGCTACCTAGCCTAAAACTTT
                  CAAAAAGCCTGATTATTGTTTAAAATGAATGATCGTTCACCGGTCAGAGGACAAGTATGA
                  CAACCCAAACAGCTTCTAGTGCCAATGCCCTTGCTTCCTTTAACCAATTTTTAAGGGATG
                  TAAAGGCGATCGCCCAACCCTATTGGTATCCCACTGTATCAAATAAAAGAAGCTTTTCTG
                  AGGTTATTCGTTCCTGGGGAATGCTATCACTGCTTATCTTTTTGATTGTGGGATTAGTCG
                  CCGTCACGGCTTTTAATAGTTTTGTTAATCGTCGTTTAATTGATGTCATTATTCAAGAAA
                  AAGATGCGTCTCAATTTGCCAGTACATTAACTGTCTATGCGATCGGATTAATCTGTGTAA
                  CGCTGCTGGCAGGGTTCACTAAAGATATTCGCAAAAAAATTGCCCTAGATTGGTATCAAT
                  GGTTAAACACCCAGATTGTAGAGAAATATTTTAGTAATCGTGCCTATTATAAAATTAACT
                  TTCAATCTGACATTGATAACCCCGATCAACGTCTAGCCCAGGAAATTGAACCGATCGCCA
                  CAAACGCCATTAGTTTCTCGGCCACTTTTTTGGAAAAAAGTTTGGAAATGCTAACTTTTT
                  TAGTGGTAGTTTGGTCAATTTCTCGACAGATTGCTATTCCGCTAATGTTTTACACGATTA
                  TCGGTAATTTTATTGCCGCCTATCTAAATCAAGAATTAAGCAAGATCAATCAGGCACAAC
                  TGCAATCAAAAGCAGATTATAACTATGCCTTAACCCATGTTCGGACTCATGCGGAATCTA
                  TTGCTTTTTTTCGGGGAGAAAAAGAGGAACAAAATATTATTCAGCGACGTTTTCAGGAAG
                  TTATCAATGATACGAAAAATAAAATTAACTGGGAAAAAGGGAATGAAATTTTTAGTCGGG
                  GCTATCGTTCCGTCATTCAGTTTTTTCCTTTTTTAGTCCTTGGCCCTTTGTATATTAAAG
                  GAGAAATTGATTATGGACAAGTTGAGCAAGCTTCATTAGCTAGTTTTATGTTTGCATCGG
                  CCCTGGGAGAATTAATTACAGAATTTGGTACTTCAGGACGTTTTTCTAGTTATGTAGAAC
                  GTTTAAATGAATTTTCTAATGCCTTAGAAACTGTGACTAAACAAGCCGAGAATGTCAGCA
                  CAATTACAACCATAGAAGAAAATCATTTTGCCTTTGAACACGTCACCCTAGAAACCCCTG
                  ACTATGAAAAGGTGATTGTTGAGGATTTATCTCTTACTGTTCAAAAAGGTGAAGGATTAT
                  TGATTGTCGGGCCCAGTGGTCGAGGTAAAAGTTCTTTATTAAGGGCGATCGCCGGTTTAT
                  GGAATGCTGGCACTGGGCGTTTAGTGCGTCCTCCCCTAGAAGAAATTCTCTTTTTGCCCC
                  AACGTCCCTACATTATTTTGGGAACCTTACGCGAACAATTGCTGTATCCTCTAACCAATA
                  GTGAGATGAGCAATACCGAACTTCAAGCAGTATTACAACAAGTCAATTTGCAAAATGTGC
                  TAAATCGGGTGGATGACTTTGACTCCGAAAAACCCTGGGAAAACATTCTCTCCCTCGGTG
                  AACAACAACGCCTAGCCTTTGCTCGATTGTTAGTGAATTCTCCGAGTTTTACCATTTTAG
                  ATGAGGCGACCAGTGCCTTAGATTTAACAAATGAGGGGATTTTATACGAGCAATTACAAA
                  CTCGCAAGACAACCTTTATTAGTGTGGGTCATCGAGAAAGTTTGTTTAATTACCATCAAT
                  GGGTTTTAGAACTTTCTGCTGACTCTAGTTGGGAACTCTTAAGCGTTCAAGATTATCGCC
                  TTAAAAAAGCGGGAGAAATGTTTACTAATGCTTCGAGTAACAATTCCATAACACCCGATA
                  TTACTATCGATAATGGATCAGAACCAGAAATAGTCTATTCTCTTGAAGGATTTTCCCATC
                  AGGAAATGAAACTATTAACAGACCTATCACTCTCTAGCATTCGGAGTAAAGCCAGTCGAG
                  GGAAGGTGATTACAGCCAAGGATGGTTTTACCTACCTTTATGACAAAAATCCTCAGATAT
                  TAAAGTGGCTCAGAACTTAA

Claims

1-19. (canceled)

20. A nucleic acid encoding a peptide spacer sequence (SP), wherein

a. the peptide sequence comprises at least 4 glycine amino acids per single repeat unit (SRU), or

b. at least five proline and/or leucine amino acids per single repeat unit (SRU),

c. a SRU within the SP is between 7 and 15 amino acids in length, and

d. the SP comprises between 2 and 10 SRUs.

21. The nucleic acid according to claim 20, encoding a peptide SRU with a sequence as shown in SEQ ID NO. 20 or SEQ ID NO. 21.

22. The nucleic acid according to claim 21, with a sequence as shown in SEQ ID NO. 43 or SEQ ID NO. 44.