Patent Publication Number: US-2003228569-A1

Title: Methods and compositions for high yield production of eukaryotic proteins

Description:
[0001] This application claims priority to U.S. provisional application Serial No. 60/081,989, filed Apr. 16, 1998, and the 60/081,989 application is herein incorporated by this reference in its entirety. 
    
    
     [0002] This invention was made with government support under grant number DK46205 awarded by the National Institute of Diabetes and Digestive and Kidney Diseases and grant number GM15431 awarded by the National Institute of General Medical Sciences of the National Institutes of Health. The government has certain rights in the invention. 
    
    
     
       BACKGROUND OF THE INVENTION  
       [0003] 1. Field of the Invention  
       [0004] The present invention relates to compositions and methods for the high yield production of eukaryotic proteins and in particular membrane proteins, by expression of recombinant vectors designed for such high yield production in bacterial cells.  
       [0005] 2. Background Art  
       [0006] Certain classes of eukaryotic, prokaryotic and viral proteins, including membrane proteins, needed in large quantities for therapeutic uses as well as for biochemical and structural studies, have proven difficult to express in recombinant systems in sufficient yields. This is particularly difficult for eukaryotic proteins with multiple membrane spanning regions including, but not limited to, G-protein coupled receptors (GPCRs) and ion channels derived from eukaryotic cells (Goeddel, 1990).  
       [0007] Eukaryotic membrane proteins have been expressed in a number of eukaryotic systems including mammalian cells, baculovirus systems [up to 55 pmol/mg of protein (125 μg/L of culture); Loisel et al., 1997] and yeast cells (up to 14 pmol/mg membrane protein; Sander et al., 1994). However, none of these approaches has proven successful for the production of large quantities of purified eukaryotic proteins.  
       [0008] Furthermore, although a number of reports in the literature describe expression of eukaryotic membrane proteins such as GPCRs in prokaryotic cells (e.g.,  E. coli ), none of these systems has proven capable of producing high levels of an intact eukaryotic protein (Table I). These bacterial cell systems have produced GPCRs in amounts of approximately several hundred receptor molecules per cell, with none of the systems producing greater than 300 receptors per cell, which corresponds to approximately 5 μg protein per liter of bacterial culture.  
               TABLE 1                          Expression levels of β adrenergic receptor in  E. coli                               Leader Sequence   Expression level                       LamB   33 to 225 receptors/cell (Chapot et al. 1990)           β-galactosidase   25 receptors/cell (Marullo et al., 1988)           none   200 receptors/levels (Breyer et al. 1990)                      
 
       [0009] The present invention overcomes previous shortcomings associated with high yield production of eukaryotic proteins by providing compositions and methods for producing eukaryotic proteins and in particular, membrane proteins, in high yield (i.e., at least 100 μg protein/L of culture), for use in biochemical and structural studies and as therapeutic agents. 
     
    
    
     BRIEF DESCRIPTION OF THE DRAWING  
     [0010]FIG. 1. Western blot analysis of PGE 2 EP 2 -cI-fusion proteins produced from nucleic acid constructs comprising: no leader sequence (cI 0 ); a leader sequence consisting of amino acids 1-15 of the lambda cI repressor protein (cI- 1-15 ); a leader sequence consisting of amino acids 1-22 of the lambda cI repressor protein (cI- 1-22 ); a leader sequence consisting of amino acids 1-36 of the lambda cI repressor protein (cI 1-36 ); and a leader sequence consisting of amino acids 1-76 of the lambda cI repressor protein (cI 1-76 ). The PGE 2 EP 2 -cI-fusion proteins were produced from a construct having a T7 promoter, nucleic acid encoding the leader sequences as described above and a nucleic acid encoding the PGE 2 EP 2  protein. Proteins in nitrocellulose were blotted with an affinity-purified sheep anti-PGE 2 EP 2  antibody and a secondary anti-sheep antibody conjugated to horse radish peroxidase and reacted with substrate according to standard methods to produce a luminescent reaction product.  
     SUMMARY OF THE INVENTION  
     [0011] The present invention provides an isolated nucleic acid comprising a first nucleotide sequence encoding an amino acid sequence comprising at least three positively charged amino acid residues, positioned upstream and in frame with a second nucleotide sequence encoding a protein.  
     [0012] In addition, the present invention provides an isolated nucleic acid comprising a first nucleotide sequence encoding a DNA binding protein, positioned upstream and in frame with a second nucleotide sequence encoding a protein.  
     [0013] An isolated nucleic acid is also provided which comprises a first nucleotide sequence encoding a bacteriophage lambda repressor protein, positioned upstream and in frame with a second nucleotide sequence encoding a protein.  
     [0014] Further provided in this invention is an isolated nucleic acid having the nucleotide sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, and SEQ ID NO:6.  
     [0015] The present invention further provides a method of producing a eukaryotic protein in a bacterial cell comprising: a) introducing the expression vector of this invention, wherein the second nucleotide sequence encodes a eukaryotic protein into the bacterial cell; and b) culturing the bacterial cell under conditions whereby the second nucleotide sequence of the expression vector is expressed to produce the eukaryotic protein.  
     [0016] A method of producing a eukaryotic protein in a bacterial cell in high yield is also provided, comprising: a) introducing the expression vector of this invention, wherein the second nucleotide sequence encodes a eukaryotic protein into the bacterial cell; and b) culturing the bacterial cell under conditions whereby the second nucleotide sequence of the expression vector is expressed to produce the eukcaryotic protein in high yield.  
     [0017] Additionally, the present invention provides a method of producing a eukaryotic integral membrane protein in a bacterial cell comprising: a) introducing the expression vector of this invention, wherein the second nucleotide sequence encodes a eukaryotic integral membrane protein into the bacterial cell; and b) culturing the bacterial cell under conditions whereby the second nucleotide sequence of the expression vector is expressed to produce the eukaryotic integral membrane protein.  
     [0018] Furthermore, the present invention provides a method of producing a eukaryotic G-protein coupled receptor protein in a bacterial cell comprising: a) introducing the expression vector of this invention, wherein the second nucleotide sequence encodes a eukaryotic G-protein coupled receptor protein, into the bacterial cell; and b) culturing the bacterial cell under conditions whereby the second nucleotide sequence of the expression vector is expressed to produce the eukaryotic G-protein coupled receptor protein.  
     [0019] Additionally provided is a method of producing a eukaryotic ion channel protein in a bacterial cell comprising: a) introducing the expression vector of this invention, wherein the second nucleotide sequence encodes a eukaryotic ion channel protein into the bacterial cell; and b) culturing the bacterial cell under conditions whereby the second nucleotide sequence of the expression vector is expressed to produce the eukaryotic ion channel protein.  
     [0020] The present invention also provides a method of producing a rabbit prostaglandin (PG) E 2  EP 3  receptor protein in a bacterial cell comprising: a) introducing the expression vector of this invention, wherein the second nucleotide sequence encodes the rabbit prostaglandin E 2  EP 3  receptor protein, into the bacterial cell; and b) culturing the cell under conditions whereby the second nucleotide sequence of the expression vector is expressed to produce the rabbit prostaglandin E 2  EP 3  receptor protein.  
     [0021] The present invention further provides a method of producing a human prostaglandin E 2  EP 2  receptor protein in a bacterial cell comprising: a) introducing the expression vector of this invention, wherein the second nucleotide sequence encodes the human prostaglandin E 2  EP 2  receptor protein, into the bacterial cell; and b) culturing the bacterial cell under conditions whereby the second nucleotide sequence of the expression vector is expressed to produce the human prostaglandin E 2  EP 2  receptor protein.  
     [0022] Also provided is a method of producing a human chemokine receptor CCR-5 protein in a bacterial cell comprising: a) introducing the expression vector of this invention, wherein the second nucleotide sequence encodes the human chemokine receptor CCR-5 protein, into the bacterial cell; and b) culturing the bacterial cell under conditions whereby the second nucleotide sequence of the expression vector is expressed to produce the human chemokine receptor CCR-5 protein.  
     [0023] In addition, the present invention provides a method of producing a human β 2  adrenergic receptor protein in a bacterial cell comprising: a) introducing the expression vector of this invention, wherein the second nucleotide sequence encodes the human β 2  adrenergic receptor protein, into the bacterial cell; and b) culturing the bacterial cell under conditions whereby the second nucleotide sequence of the expression vector is expressed to produce the human β 2 , adrenergic receptor protein.  
     [0024] The present invention further provides a method of producing a rat renal outer medullary K +  channel protein in a bacterial cell comprising: a) introducing the expression vector of this invention, wherein the second nucleotide sequence encodes the rat renal outer medullary K +  channel protein, into the bacterial cell; and b) culturing the bacterial cell under conditions whereby the second nucleotide sequence of the expression vector is expressed to produce the rat renal outer medullary K +  channel protein.  
     [0025] Finally provided is a method of producing a human small G-protein rho protein in a bacterial cell comprising: a) introducing the expression vector of this invention, wherein the second nucleotide sequence encodes the small G-protein rho protein, into the bacterial cell; and b) culturing the bacterial cell under conditions whereby the second nucleotide sequence of the expression vector is expressed to produce the small G-protein rho protein.  
     [0026] Various other objectives and advantages of the present invention will become apparent from the following detailed description.  
     DETAILED DESCRIPTION OF THE INVENTION  
     [0027] As used herein, “a” or “an” can mean multiples. For example, “a cell” can mean at least one cell or more than one cell.  
     [0028] The present invention provides an isolated nucleic acid comprising a first nucleotide sequence encoding an amino acid sequence comprising at least three positively charged amino acid residues, positioned upstream and in frame with a second nucleotide sequence encoding a protein. As used herein, an “amino acid sequence comprising at least three positively charged residues” means an amino acid sequence having at least three and possibly more than three positively charged residues (e.g. arginine, lysine etc.) which can be consecutive, closely spaced, or randomly spaced. The amino acid sequence comprising at least three positively charged residues is the “leader sequence” of the fusion protein made by the methods of this invention. Neither the leader sequence nor the overall protein sequence (i.e., the leader sequence and the protein sequence together) need have a net positive charge, i.e., a pI value&gt;7. The leader sequence can be as short as five amino acids (aa) long (e.g., 15 aa), although longer sequences (e.g., about 36 to 76 amino acids) are preferred. For example, the leader sequence of the fusion protein of this invention can comprise a nucleic acid encoding an amino acid sequence of a DNA binding protein, such as the bacteriophage lambda repressor protein.  
     [0029] Thus, the present invention further provides an isolated nucleic acid comprising a first nucleotide sequence encoding a DNA binding protein, positioned upstream and in frame with a second nucleotide sequence encoding a protein. As used herein, a “DNA binding protein” means a protein, which in its native setting, binds DNA and regulates its function. The DNA binding protein can be selected from the group consisting of eukaryotic DNA binding proteins, prokaryotic DNA binding proteins and bacteriophage-derived DNA binding proteins. For example, the DNA binding proteins of this invention can include, but are not limited to, bacteriophage DNA binding proteins such as lambda (λ) repressor, λ cro repressor, phage P22 arc repressor and phage P22 nmt repressor; bacterial DNA binding proteins such as the lac repressor and the trp repressor; eukaryotic binding proteins such as the yeast gal 4 protein; and mammalian transcription factors such as fos and jun as well as histones, transcriptional activators such as CREB and any other DNA binding protein now known or later identified.  
     [0030] It is also appreciated by one of skill in the art that DNA binding proteins can include fragments which retain at least three positively charged residues.  
     [0031] Also provided is an isolated nucleic acid comprising a first nucleotide sequence encoding a bacteriophage lambda repressor protein (having 236 amino acids, as shown in SEQ ID NO:9), positioned upstream and in frame with a second nucleotide sequence encoding a protein. In addition, the first nucleotide sequence of the nucleic acid of this invention can encode the N-terminal domain of the bacteriophage lambda repressor protein (having amino acids 1-92, as shown in SEQ ID NO:10), amino acids 1-76 (SEQ ID NO:11) of the bacteriophage lambda repressor protein, or at least 15 contiguous amino acids of the N-terminal domain of the bacteriophage lambda repressor protein.  
     [0032] The λ repressor protein is a product of the cl gene of bacteriophage λ. This gene encodes a protein of 236 amino acids organized into two domains, and N-terminal DNA binding domain consisting of amino acids 1-92 and a C-terminal domain consisting of amino acids ˜114-236 (Sauer, 1978; Sauer et al., 1979).  
     [0033] In the nucleic acid of this invention as described above, the second nucleotide sequence can encode any protein which can be produced exogenously in a bacterial protein expression system. For example, the protein of this invention can be a viral protein, a prokaryotic protein or a eukaryotic protein. Viral proteins encoded by the second nucleotide sequence of the nucleic acid of this invention can include, but are not limited to, a homolog of the G-coupled receptor protein from cytomeaglovirus, herpesvirus 6, herpesvirus 7, Kaposi&#39;s sarcoma-associated herpesvirus (human herpesvirus 8), herpesvirus saimiri (e.g., the gene product ot ECRF3) human immuodeficiency virus (HIV) proteins gp120 and gp41, measles virus F protein, influenza hemagglutinin protein and herpesvirus B and H proteins. Prokaryotic proteins encoded by the second nucleotide sequence of the nucleic acid of this invention can include, but are not limited to, diacylglycerol kinase bacterial membrane protein and the lamB gene product. Eukaryotic proteins encoded by the second nucleotide sequence of the nucleic acid of the present invention can include, but are not limited to eukaryotic proteins selected from the group consisting of integral membrane proteins, G-protein coupled receptor (GPCR) proteins and ion channel proteins. Integral membrane proteins are proteins which have at least one hydrophobic amino acid sequence which passes through the membrane lipid bilayer as a transmembrane region or domain. GPCRs are a superfamily of integral membrane proteins which are widely distributed in eukaryotic cells and consist of seven transmembrane domains interconnected by a series of peptide loops. In their native environment, these proteins bind ligand from their exofacial surface and transmit signal to the intracellular side via heterotrimeric guanine-nucleotide binding proteins (G-proteins). Ion channel proteins are integral membrane proteins which, in their native environment, the plasma membrane of virtually all cell types, form a pore in the lipid bilayer which allows the selective passage of one or more ions, either into or out of the cell.  
     [0034] Furthermore, the second nucleotide sequence of the nucleic acid of this invention can encode a protein selected from the group consisting of rabbit prostaglandin E 2 EP 3  receptor protein, human prostaglandin E 2 EP 2  receptor protein, human chemokine receptor CCR-5 protein, human β 2  adrenergic receptor protein, rat renal outer medullary K +  channel protein and human small G-protein rho.  
     [0035] The present invention additionally provides an isolated nucleic acid having the nucleotide sequence selected from the group consisting of SEQ ID NO:1 (plasmid pLJM5.22His encoding the cI-77A his fusion protein), SEQ ID NO:2 (plasmid pCK2.5 HTL encoding cI-EP 2  his-thrombin-lambda repressor C terminal domain aa 82-236), SEQ ID NO:3 (plasmid pSD1.63 his encoding cI-CCR5 his), SEQ ID NO:4 plasmid pSD1.18his encoding cI-βAR his), SEQ ID NO:5 (plasmid pSD1.134his encoding cI-ROMK; his), and SEQ ID NO:6 (plasmid pSD2.46his encoding cI-rho his).  
     [0036] As used herein, “nucleic acid” refers to single- or double-stranded molecules which may be DNA, comprising two or more nucleotides comprised of the nucleotide bases A, T, C and G, or RNA, comprised of the bases A, U (substitute for T), C and G. The nucleic acid may represent a coding strand or its complement. Thus, the present invention also provides nucleic acids complementary to, or capable of, hybridizing with the nucleic acids of this invention. The nucleic acid of this invention may be a naturally occurring nucleic acid or the nucleic acid may be a synthetic nucleic acid sequence which contains alternative codons which encode the same amino acid as that which is found in a naturally occurring sequence (Lewin, 1994). Furthermore, the nucleic acids of this invention can include codons which encode amino acids which represent conservative substitutions of amino acids that do not alter the function of the protein, as are well known in the art.  
     [0037] As used herein, the term “isolated” means a nucleic acid separated or substantially free from at least some of the other components of the naturally occurring organism, for example, the cell structural components commonly found associated with nucleic acids in a cellular environment and/or other nucleic acids. The isolation of nucleic acids can therefore be accomplished by techniques such as cell lysis followed by phenol plus chloroform extraction, followed by ethanol precipitation of the nucleic acids (Michieli et al.,1996). The nucleic acids of this invention can be isolated from cells according to methods well known in the art. Alternatively, the nucleic acids of the present invention can be synthesized according to standard protocols well described in the literature.  
     [0038] The nucleic acid of this invention can be part of a recombinant nucleic acid comprising any combination of restriction sites and/or functional elements as are well known in the art which facilitate molecular cloning, expression, post-translational modifications and other recombinant DNA manipulations. For example, the nucleic acid of this invention encodes a leader sequence fused to a protein sequence to produce a fusion protein from which the leader sequence can be cleaved to yield only the protein sequence. Thus, nucleotide sequences which encode amino acid sequences which provide for the enzymatic or chemical cleavage of the leader peptide from the mature polypeptide, as well as regulatory sequences which allow temporal regulation of expression of the nucleic acid of this invention can also be included in the nucleic acid of this invention. Thus, the present invention further provides a recombinant nucleic acid comprising the nucleic acid of the present invention. In particular, the present invention provides a vector comprising the nucleic acid of this invention and a cell comprising the vector of this invention.  
     [0039] The vector of this invention can be an expression vector which contains all of the genetic components required for expression of the nucleic acid in cells into which the vector has been introduced, as are well known in the art. The expression vector can be a commercial expression vector or it can be constructed in the laboratory according to standard molecular biology protocols.  
     [0040] The vector of this invention is introduced into a bacterial cell under conditions whereby the resulting stable transformants maintain the vector, as are well known in the art and as described in the Examples provided herein.  
     [0041] The vector of this invention is introduced into a bacterial cell according to standard procedures for introducing nucleic acid into prokaryotes, as are well known in the art. There are numerous  E. coli  expression vectors known to one of ordinary skill in the art useful for the expression of proteins in prokaryotic systems. Other microbial hosts suitable for use include bacilli, such as  Bacillus subtilus  and other enterobacteria, such as Salmonella, Serratia and various Pseudomonas species. Expression vectors for prokaryotic systems typically contain expression control sequences compatible with the host cell (e.g., an origin of replication) and an antibiotic resistance marker to provide for the growth and selection of the expression vector in a bacterial host. In addition, any number of a variety of well-known promoters can be present, such as the T7 promoter system, the lactose promoter system, a tryptophan (Trp) promoter system, a beta-lactamase promoter system, or a promoter system from phage lambda. The promoters will typically control expression, optionally with an operator sequence and have ribosome binding site sequences for example, for initiating and completing transcription and translation. The vector can also contain expression control sequences, enhancers that may regulate the transcriptional activity of the promoter, appropriate restriction sites to facilitate cloning of inserts adjacent to the promoter and other necessary information processing sites, such as RNA splice sites, polyadenylation sites and transcription termination sequences as well as any other sequence which may facilitate the expression of the inserted nucleic acid.  
     [0042] The nucleic acid in the vector of this invention can be expressed in cells after the nucleotide sequences have been operably linked to, i.e., positioned, to ensure the functioning of an expression control sequence. These expression vectors are typically replicable in the cells either as episomes or as an integral part of the cell&#39;s chromosomal DNA. Commonly, expression vectors can contain selection markers, e.g., tetracycline resistance, ampicillin resistance, kanamycin resistance or chlormaphenicol resistance, etc., to permit detection and/or selection of those bacterial cells transformed with the desired nucleic acid sequences (see, e.g., U.S. Pat. No. 4,704,362).  
     [0043] Thus, the present invention provides a method of producing a eukaryotic protein in a bacterial cell comprising: a) introducing the expression vector of this invention, wherein the second nucleotide sequence encodes a eukaryotic protein into the bacterial cell; and b) culturing the bacterial cell under conditions whereby the second nucleotide sequence of the expression vector is expressed to produce the eukaryotic protein.  
     [0044] A method of producing a eukaryotic protein in a bacterial cell in high yield is also provided, comprising: a) introducing the expression vector of this invention, wherein the second nucleotide sequence encodes a eukaryotic protein, into the bacterial cell; and b) culturing the bacterial cell under conditions whereby the second nucleotide sequence of the expression vector is expressed to produce the eukaryotic protein in high yield. As used herein, “high yield” means that the protein is produced in an amount which is at least, and preferably is greater than, 100 μg/liter of bacterial culture. More preferably, high yield means the protein is produced in an amount which is at least 0.5 mg/liter of culture and most preferably, the protein is produced in an amount which is at least 2.0 mg/liter of culture.  
     [0045] In addition, the present invention provides a method of producing a eukaryotic integral membrane protein in a bacterial cell comprising: a) producing the expression vector of this invention, wherein the second nucleotide sequence encodes a eukaryotic integral membrane protein; b) introducing the vector into the bacterial cell; and c) culturing the bacterial cell under conditions whereby the second nucleotide sequence of the expression vector is expressed to produce the eukaryotic integral membrane protein.  
     [0046] The present invention also provides a method of producing a eukaryotic G-protein coupled receptor protein in a bacterial cell comprising: a) introducing the expression vector of this invention, wherein the second nucleotide sequence encodes a eukaryotic G-protein coupled receptor protein, into the bacterial cell; and b) culturing the bacterial cell under conditions whereby the second nucleotide sequence of the expression vector is expressed to produce the eukaryotic G-protein coupled receptor protein.  
     [0047] Additionally, the present invention provides a method of producing a eukaryotic ion channel protein in a bacterial cell comprising: a) introducing the expression vector of this invention, wherein the second nucleotide sequence encodes a eukaryotic ion channel protein, into the bacterial cell; and b) culturing the bacterial cell under conditions whereby the second nucleotide sequence of the expression vector is expressed to produce the eukaryotic ion channel protein.  
     [0048] Furthermore, the present invention provides methods for the production of specific eukaryotic proteins. In particular, the present invention provides a method of producing a rabbit prostaglandin E 2  EP 3  receptor protein in a bacterial cell comprising: a) introducing the expression vector of this invention, wherein the second nucleotide sequence encodes the rabbit prostaglandin E 2  EP 3  receptor protein, into the bacterial cell; and b) culturing the cell under conditions whereby the second nucleotide sequence of the expression vector is expressed to produce the rabbit prostaglandin E 2  EP 3  receptor protein. The rabbit PGE 2 EP 3  receptor protein is a member of the family of GPCRs, which in its native environment (e.g., the plasma membrane of cells in the kidney, stomach and adrenal glands, among others), binds prostaglandin E 2  and elicits intracellular signals (Breyer, et al., 1994) on the intracellular side via heterotrimeric guanine-nucleotide binding proteins (G-proteins).  
     [0049] Also provided is a method of producing a human prostaglandin E 2  EP 2  receptor protein in a bacterial cell comprising: a) introducing the expression vector of this invention, wherein the second nucleotide sequence encodes the human prostaglandin E 2  EP 2  receptor protein, into the bacterial cell; and b) culturing the bacterial cell under conditions whereby the second nucleotide sequence of the expression vector is expressed to produce the human prostaglandin E 2  EP 2  receptor protein. The human PGE 2  EP 2  receptor is a member of the family of GPCRs which, in its native environment (e.g., the plasma membrane of cells in the lung, uterus and blood cells, among others), binds prostaglandin E 2  and elicits intracellular signals on the intracellular side via heterotrimeric guanine-nucleotide binding proteins (G-proteins).  
     [0050] Further provided is a method of producing a human chemokine receptor CCR-5 protein in a bacterial cell comprising: a) introducing the expression vector of this invention, wherein the second nucleotide sequence encodes the human chemokine receptor CCR-5 protein, into the bacterial cell; and b) culturing the bacterial cell under conditions whereby the second nucleotide sequence of the expression vector is expressed to produce the human chemokine receptor CCR-5 protein. The human chemokine receptor CCR-5 (alternatively named CC-CKR5) is a member of the family of GPCRs which, in its native environment (the plasma membrane of T cells and macrophages, among others), binds chemokine peptide hormones on the exofacial surface of the cell and elicits intracellular signals on the intracellular side of the plasma membrane via heterotrimeric guanine-nucleotide binding proteins (G-proteins). In addition, CCR-5 is utilized by the human immunodeficiency virus (HIV) virus as a co-receptor which facilitates viral entry into the host cell during the pathogenesis of viral infection.  
     [0051] Production of the chemokine receptor protein, CCR-5, in high yield provides for a number of therapeutic uses. For example, the CCR-5 protein can be used as an immunogen to develop autoantibodies to the CCR-5 protein. This active immunization would then inhibit HIV entry into target cells expressing the CCR-5 receptor. Alternatively, the CCR-5 protein can be used for passive immunization wherein an animal (e.g., a horse) can be immunized with the CCR-5 fusion protein of this invention and the resulting antiserum collected and purified. The horse Ig anti-CCR-5 fusion protein fraction can then be administered to humans to inhibit HIV entry into target cells expressing the CCR-5 receptor. A third use of the CCR-5 protein can be to administer the CCR-5 protein of this invention into a subject infected with HIV in an amount which can bind to HIV in the subject and inactivate it.  
     [0052] In addition, the present invention provides a method of producing a human β 2  adrenergic receptor protein in a bacterial cell comprising: a) introducing the expression vector of this invention, wherein the second nucleotide sequence encodes the human β 2  adrenergic receptor protein, into the bacterial cell; and b) culturing the bacterial cell under conditions whereby the second nucleotide sequence of the expression vector is expressed to produce the human β 2  adrenergic receptor protein. The human β 2  adrenergic receptor is a member of the family of GPCRs which, in its native environment (the plasma membrane of cells in the heart, lungs, blood vessels, intestine and other organs and tissues), binds epinephrine and its natural and synthetic analogs on their exofacial surface and elicits intracellular signals on the intracellular side via heterotrimeric guanine-nucleotide binding proteins (G-proteins).  
     [0053] Furthermore, the present invention provides a method of producing a rat renal outer medullary K +  channel (ROMK) protein in a bacterial cell comprising: a) introducing the expression vector of this invention, wherein the second nucleotide sequence encodes the rat renal outer medullary K +  channel protein, into the bacterial cell; and b) culturing the bacterial cell under conditions whereby the second nucleotide sequence of the expression vector is expressed to produce the rat renal outer medullary K +  channel protein. ROMK is a member of the family of ion channel proteins which, in its native environment (the plasma membrane of cells in the kidney, brain, heart and stomach), allows the selective secretion of potassium ions from the intracellular milieu.  
     [0054] The present invention also provides a method of producing a human small G-protein rho protein in a bacterial cell comprising: a) producing the expression vector of this invention, wherein the second nucleotide sequence encodes the small G-protein rho protein; b) introducing the vector into the bacterial cell; and c) culturing the bacterial cell under conditions whereby the second nucleotide sequence of the expression vector is expressed to produce the small G-protein rho protein. The protein rho is a member of the family of “small GTP binding proteins” which, in its native environment, is a membrane associated protein but is not an integral membrane protein i.e., it does not contain any transmembrane regions.  
     [0055] The methods of this invention can further comprise the step of isolating and purifying the protein according to methods well known in the art and as described herein (see, e.g., Sambrook et al.). Additionally, for all of the methods of the present invention, the eukaryotic protein which is produced can be produced in high yield, as defined herein.  
     [0056] The proteins produced by the methods of this invention can be cleaved from the leader sequence as described herein, re-folded and tested for functionality. For example, the proteins of this invention can be refolded according to methods well known in the art for refolding membrane proteins (see, e.g., Braiman et al., 1987). Briefly, the inclusion bodies containing the fusion protein of this invention can be isolated from the bacterial cells in which they were produced and solubilized in buffer containing 0.2% sodium dodecyl sulfate (SDS). The fusion proteins can then be mixed with buffer containing dimyristoyl phosphatidyl choline (DMPC) and CHAPS detergent, to allow for the renaturation of the detergent-solubilized protein.  
     [0057] The proteins of this invention can be tested for functionality by a variety of methods. For example, the presence of antigenic epitopes and ability of the proteins to bind ligands can be determined by Western blot assays, fluorescence cell sorting assays, immunoprecipitation, immunochemical assays and/or competitive binding assays, as well as any other assay which measures specific binding activity.  
     [0058] For ion channel proteins of this invention, the proteins can be reconstituted into artificial lipid bilayers (so-called black lipid bilayers) according to methods well known in the art. The ability of the ion channel to function can then be tested electrophysiologically, using direct current/voltage measurements according to well known methods (Zweifach et al., 1991). Alternatively, the ion channel can be incorporated into liposomes according to standard methods, and activity can be monitored by the ability of ion fluxes to change the fluorescence of indicator dyes contained within the liposomes, as known in the art (Le Caherec et al., 1996; Zeidel et al., 1992).  
     [0059] GPCRs can be tested for functionality by testing them for the ability to bind radiolabeled ligand in a specific and saturable manner, as is well known in the art (Limbird, 1996). Moreover, activation of GPCRs leads to a concomitant increase in enzymatic activity by GTP binding proteins (increased turnover of GTP to GDP). The GPCRs of this invention can also be reconstituted with purified heteromeric G-proteins in liposomes and the rate of GTP hydrolysis upon addition of the cognate agonist can be measured according to well known methods. An increase in rate indicates a properly folded and functioning protein.  
     [0060] The proteins of this invention can be used in a number of practical applications including, but not limited to:  
     [0061] 1. Immunization with recombinant host protein antigen as a viral/pathogen antagonist.  
     [0062] 2. Production of membrane proteins for diagnostic or screening assays.  
     [0063] 3. Production of membrane proteins for biochemical studies.  
     [0064] 4. Production of membrane protein for structural studies.  
     [0065] 5. Antigen production for generation of antibodies for immuno-histochemical mapping, including mapping of orphan receptors and ion channels.  
     [0066] In particular, the proteins of this invention can be used as immunogens in vaccine protocols and as antigens for the production of monoclonal and polyclonal antibodies according to methods standard in the art (see, e.g., Harlow and Lane, 1988). Additionally, the proteins of this invention can be used in diagnostic and screening assays to detect the presence of a ligand which binds the protein of this invention, in a biological sample from a subject. Such assays can be carried out by detecting the binding of the protein with a ligand in the sample. Such detection methods can include identifying the formation of a protein/ligand complex with detectable antibodies which specifically bind the protein and/or the ligand as well as by competitive binding assays as are well known in the art.  
     [0067] The present invention is more particularly described in the following examples which are intended as illustrative only since numerous modifications and variations therein will be apparent to those skilled in the art. 
    
    
     EXAMPLES  
     Example 1  
     Plasmid pLJM5-22H-Fusion of Rabbit EP 3  Receptor With the N-terminal Domain of λ Repressor.  
     [0068] Construction of cI-77A fusion  E. coli  expression vector—Plasmid pLJM5-22H (SEQ ID NO:1) consists of the first 231 bp of a semi-synthetic cI gene (Breyer and Sauer, 1989a), encoding amino acids 1-76 (SEQ ID NO:11), fused to the N-terminus of the EP 3  receptor 77A splice variant. It should be noted that the numbering convention for the λ-repressor protein designates serine encoded at codon 2 of the cI gene as position 1 because the N-terminal methionine is cleaved in the mature λ-repressor protein. It is not known whether the cleavage of the initiator methionine occurs in the fusion proteins, however the convention designating serine as amino acid 1 is utilized for the various fusion proteins described below.  
     [0069] The C-terminus of the 77A protein was modified to remove the stop codon, and an XhoI restriction site was introduced immediately 3′ to the coding region allowing a C-terminal fusion. The vector, ptac promoter and synthetic cI gene sequences were derived from the plasmid pRB200 (Breyer and Sauer, 1989a). Internal EcoRI and XhoI sites in the synthetic cI gene were digested and the 77A cDNA was inserted as follows to produce the plasmid: an EcoRI/NdeI adapter pair [5′-AATTCGCAGCTCA-3′ (SEQ ID NO:16) and 5′-TATGAGCTGCG 3′ (SEQ ID NO:17)] was used to fuse the initiator methionine of 77A from plasmid pRC/CMV 77A wt (Audoly and Breyer, 1997) in frame with amino acid 76 of the cI sequence. The 3′ end of the 77A cDNA was modified by PCR, removing the TGA stop codon and the 3′ untranslated region. An XhoI restriction site was simultaneously introduced allowing fusion to a C-terminal 6×-his tag. The sequence of the sense primer, 5′-ACA TCA GTT GAG CAC TGC-3′ (SEQ ID NO:18) lies within the 77A coding region. The antisense primer 5′-CCT CGA GGC TTG CTG ATA AGG ACG AGC-3′ (SEQ ID NO:19) lies at the 3′ end of the coding region. PCR was performed with Vent DNA polymerase for 30 cycles of 94° C. for 15 sec, 51° C. for 15 sec and 72° C. for 30 sec. The PCR fragment was digested with BsmI (a restriction site internal to 77A) and XhoI and ligated into the 77A cDNA backbone reconstituting the full length 77A cDNA with an XhoI site at the 3′ end of the coding region. This plasmid was digested at the XhoI site and a HindIII site 3′ to the EP coding region. The his tag linker oligonucleotide sequences 5′ TCG AGG CAC CAT CAC CAC CAC CAC TGA A 3′ (SEQ ID NO:20) and 5′ AGC TTT CAG TGG TGG TGG TGA TGG TGC C 3′ (SEQ ID NO:21) oligonucleotides encoding a 6× his tag were ligated to the backbone, allowing fusion to a C-terminal 6× his tag used for affinity purification.  
     [0070] The NT (No-Tail) EP 3  receptor expression construct, which lacks the C-terminal sequence encoded by the alternatively spliced variable sequence of the 77A splice variant, was generated by PCR using the 77A cDNA as a template. The receptor sequence ends at Q 355 , 10 residues distal to the end of transmembrane VII, and is fused to the 6× his tag at the C-terminus. A PCR fragment was generated using a sense internal primer at nt 532 of the coding region (5′-TGG CTG GCA GTG CTC GCC-3′) (SEQ ID NO:22) and a downstream primer (5′-TCA CCT CGA GGC CTG GCA AAA CTT CCG AAG-3′) (SEQ ID NO:23) which inserts an XhoI site immediately distal to nt 1065 (Q 355 ), allowing amplification of a 534 bp fragment. The PCR was performed with Vent DNA polymerase for 35 cycles of 98° C. for 15 sec, 57° C. for 15 sec and 72° C. for 60 sec. This PCR product was digested at internal SacI and XhoI restriction sites and the resulting fragment was subcloned into the SacI and XhoI sites in expression vector pLJM5-22H to yield the plasmid pLJM6-09 (SEQ ID NO:7) which expresses the cI-NT-his fusion protein.  
     [0071] Plasmid pLJM5-42T (SEQ ID NO:8) expresses a third variant of the 77A protein, cI-77A-TL. In addition to the N-terminal cI 1-76  fusion, the cI-77A-TL construct (for Thrombin-Lambda C-terminal fusion) is fused to amino acids 82-236 of the cI gene utilizing a thrombin cleavage site linker between the C-terminal sequence of 77A and the lambda C-terminal fusion. The thrombin cleavage sequence LVPRGS (SEQ ID NO:22), allows cleavage of the C-terminal fusion from the purified recombinant protein. This construct was made by inserting the oligonucleotide pair 5′ TCG ACC CTG GTG CCA CGC GGA TCC GT 3′ (SEQ ID NO:37) and 5′ TCG AAC GGA TCC GCG TGG CAC CAG GC 3′ (SEQ ID NO:24) into the XhoI site at the 3′ end of the 77A fragment outlined above. This allows an in-frame fusion of the 77A receptor to the thrombin cleavage sequence followed by amino acids 82-236 of the cI gene. This thrombin cleavage sequence or similar sequences (e.g. enterokinase) can be inserted between the N-terminal cI- 1-76  fusion as well, allowing the isolation of protein products which lack the fusion peptides.  
     [0072] Induction of cI-EP 3  fusion protein expression.  E. coli  strain DH5α cells transformed with various expression plasmids was grown in 2×LB medium containing 100 μg/ml of ampicillin (2×LBA100). Cells were grown in 2×LBA100 with shaking at 37° C. until the culture reached an A 600  of 0.8. Protein expression was induced by addition of 1 mM isopropyl β-thiogalactoside (IPTG), followed by a further incubation at 30° C. for 5 hours. Cells were harvested by centrifugation at 2,500×g, flash frozen in liquid nitrogen and stored at −80° C.  
     [0073] Protein Purification by Ni-NTA Column—Bacterial cell pellets were resuspended in Buffer 1 (50 mM Tris Cl, 150 mM Na Cl, 0.1% NaN 3 , 10 mM CHAPS 20% glycerol, 2 mM PMSF, 1.4 mM β-ME, pH 8.0) followed by sonication three times for 20 sec, on ice. The inclusion bodies were collected by centrifugation at 35,000×g for 20 min at 4° C. The supernatant was discarded and the inclusion bodies were washed two more times with Buffer 1 using the same centrifugation protocol. Washed inclusion bodies were dissolved in Buffer 2 (50 mM Tris Cl, 500 mM Na Cl, 1% NP-40, 0.5% Na deoxycholate, 2 mM PMSF, 2 M urea, 20 mM imidazole, pH 8.0). Ni-NTA agarose beads were added and incubated at 4° C. overnight on a rotary shaker. Agarose beads were collected by centrifugation at 1000×g for 2 min, and batch washed with 50 volumes of Buffer 2. After three washes, the fusion protein was batch eluted with Buffer 3 (50 mM Tris Cl, 500 mM Na Cl, 1% NP-40, 0.5% Na deoxycholate, 2 mM PMSF, 2 M urea, 100 mM imidazole, pH 8.0). Eluate was dialyzed against Buffer 2 and purified a second time on Ni-NTA beads as described above.  
     [0074] Quantitation of Protein. Purified protein was quantitated using the BCA protein assay (Pierce). Estimates of the specific content of EP 3  fusion protein in the lysate were made utilizing a “dot-blot” immuno-assay employing the 24H monoclonal antibody directed against the N-terminal domain of the fusion partner. The dot blot assay was performed as follows: Bacterial cells expressing the cl-fusion protein were induced for the required time at the appropriate temperature with 1 mM IPTG. The lysate was fractionated to obtain the following fractions: cell lysate, cytosol, washed inclusion bodies, solubilized inclusion bodies and Ni-NTA purified protein. One μl of the various purification fractions containing the expressed cI-fusion protein were “spotted” onto nitrocellulose filters and allowed to air dry. Additionally, 1 μl volumes of a range of known amounts of purified fusion protein, the concentration of which was determined by BCA assay, were spotted as a standard curve. The dried blot was then processed for immuno-detection: the blot was washed with Tween 20 containing Tris buffered Saline (TBS-T) for 10 min followed by a 1 hr blocking step using TBS-T containing 5% (w/v) skim milk. The blot was then rinsed briefly with TBS-T and incubated overnight at 4° C. with TBS-T containing 2% skim milk with an appropriate dilution of mouse 24H antibody. The blot was then washed and incubated with horseradish peroxidase-conjugated goat anti-mouse IgG. The blot was subsequently washed 3×10 min with TBS-T and a chemiluminescence reaction was performed according to the manufacturer&#39;s instructions (SuperSignal substrate, Pierce). Intensity of each dot was compared to the known standard by densitometry. Overall yield of the expression/purification was calculated by factoring in corrections for volume of each fraction.  
     [0075] Production of anti-fusion protein antibodies. The purified fusion protein of this invention can be used to raise specific antibodies (either monoclonal or polyclonal) against the expressed protein according to protocols well known in the art. In the case of the EP 3  receptor, the purified cI-77A-his protein was injected into goats. The first injection was made subcutaneously with 0.5 mg of recombinant protein in Complete Freunds&#39; adjuvant. Subsequent boost immunizations were made subcutaneously with 0.25 mg of recombinant protein in Incomplete Freunds&#39; adjuvant. Antiserum was “depleted” by incubation with  E. coli  lysate expressing the intact cI gene which had been covalently coupled to the CNBr activated Sepharose resin. This incubation removes or depletes antibodies directed against the cI fusion partner sequence as well as any antibodies raised to  E. coli  proteins from either natural infection or present as minor contaminants in the immunogen. The supernatant retains antibodies to the partner EP 3  protein. The resulting depleted antiserum may be further purified by adsorption to the purified cI-77A-his protein antigen coupled to CNBr Sepharose. Purified antibody is subsequently eluted from the resin and can be utilized, for example, in immunodetection assays to identify the EP 3  protein in native tissues or from recombinant sources.  
     [0076] Ligand Binding Studies—The fusion proteins of the present invention can be refolded as described herein and used in a variety of assays, such as ligand binding studies. For example, the refolded fusion proteins can be used to screen drugs in a variety of assays as are well known to one of skill in the art. For example, on the basis that HIV must interact with CCR-5 to gain entry into cells, substances can be screened for antiviral activity by detecting the ability of the substance to block the binding to the HIV gp120 coat protein to the CCR-5 receptor protein of this invention. Plastic microtiter plates can be coated with the CCR-5 protein and radiolabeled HIV gp120 coat protein (which can be complexed with recombinant CD-4, the co-receptor) can be added in the presence of the substance to be screened for antiviral activity. The amount of radioactive gp120 coat protein bound to the plate in the absence and presence of the substance can be determined according to standard methods. A decrease in or absence of bound gp120 as determined by quantitating the radioactive signal indicates a substance having the ability to inhibit binding of the HIV gp120 coat protein to the CCR-5 receptor protein, thereby identifying a substance potentially having antiviral activity. The substance can then be further screened for specific antiviral activity according to protocols well known in the art.  
     [0077] The protein of this invention can also be used in saturation binding isotherm experiments. For example, the recombinant membrane protein of this invention is incubated in binding buffer (25 mM KPO 4 , pH 6.2, 10 mM MgCl 2 , and 1 mM EDTA) for 2 h at 30° C. with varying concentrations of [ 3 H] PGE 2 . Nonspecific binding is determined in the presence of 50 μM unlabeled PGE 2 . Reactions are stopped by rapid filtration on Whatman GF/F glass fiber filters as described previously (Breyer, et al., 1994). Filters are washed three times with binding buffer, dried, and counted in Dupont 989 fluor.  
     [0078] Expression of cI-77A-his fusion protein—The vector pLJM5-22H has the tac promoter and the cI translation initiator sequences. The cDNA encoding the 77A splice variant of the EP 3  receptor was modified as described above to obtain an EP 3  receptor fused to a portion of the cI gene (1-76 aa) at the N-terminus and a C-terminal fusion to a 6× his tag. The resultant fusion protein of 501 aa has a predicted molecular weight 55.4 kDa. When expressed in  E. coli  this construct demonstrated high levels of expression of a protein of apparent molecular weight of 50 kDa in whole cell lysates when resolved by SDS-PAGE. Expression levels of the cI-77A-his fusion protein were estimated to be in the range of 20 mg/L of  E. coli  culture as determined by dot-blot assay as described herein.  
     [0079] A second fusion protein cI-NT (No Tail)-his, in which the C-terminal tail amino acids 356 to 411 of the 77A protein were deleted, was also constructed. The cI-NT-his protein expressed by this construct lacks the epitope to which the anti-peptide antibody was generated and thus served as a control for the anti tail-peptide antibody in Western blot experiments. A third construct fused the C-terminus of lambda repressor, residues 82-236, to the C-terminus of the cI-77A fusion via a thrombin cleavage site linker. This protein designated cI-77A-TL (Thrombin-Lambda) tested the hypothesis that the C-terminal sequence of  E. coli  expressed proteins are important determinants of protein degradation. Because lambda repressor is highly expressed in  E. coli , its C-terminus might be resistant to degradation by C-terminal directed proteases. Coomasie blue and Western blot analysis of the lysates resolved by SDS-PAGE demonstrated that the induced protein of the appropriate molecular weight was reactive with the 24H antibody for each of the EP 3  constructs tested. This 24H monoclonal Ab was generated against the N-terminal 102 amino acids of the cI gene and its recognition sequence is contained within the first 36 amino acids of the antigen (Breyer and Sauer, 1989b).  
     [0080] Western blot analysis was also performed with an antibody raised against the unique sequence in the 77A cDNA as described herein. A protein of 50 kDa was detected for the cI-77A-his lysate, and a protein of 68 kDa was detected for the cI-77A-TL but neither the cI containing lysate nor the cI-NT-his, in which the target epitope had been deleted, displayed any reactivity.  
     [0081] Purification of cI-77A-his fusion protein. When the plasmid encoding the cI-77A-his fusion protein was expressed in  E. coli , the majority of the cI-77A-his fusion protein produced was found in the insoluble fraction comprising the “inclusion bodies.” Precipitation of a protein into inclusion bodies can be advantageous because the precipitated protein is subjected to minimal proteolysis and can be recovered as a partially pure aggregate. Moreover, mammalian membrane proteins can be toxic when expressed in  E. coli  membrane and sequestration of the PGE 2 EP 3  receptor in inclusion bodies can remove some of the selective disadvantage of PGE 2 EP 3  overexpression.  
     [0082] Inclusion bodies were collected by centrifugation, which separated them from the majority of soluble protein contaminants. The inclusion bodies were washed extensively in the presence of CHAPS, with only a small loss of cI-77A-his. The washed inclusion bodies were solubilized in 2M urea in the presence of NP-40 and deoxycholate. Solubilized cI-77A-his was then purified by affinity chromatography using Ni-NTA agarose resin in buffer containing 2M urea, NP-40 and deoxycholate. After a second round of affinity purification on the Ni-NTA resin, the cI-77A-his was purified to apparent homogeneity as determined by silver staining, with a yield of approximately 10% of the initial protein expressed or approximately 2 mg protein purified per liter of bacterial culture.  
     Example 2  
     Plasmid pCK2-5HTL-Fusion of the Human EP 2  Receptor With the N-terminal Domain of λ Repressor.  
     [0083] Construction of cI-EP 2  HTL fusion  E. coli  expression vector. Plasmid pCK2-5HTL (SEQ ID NO:2) consists of the first 231 bp of a semi-synthetic cI gene (Breyer and Sauer, 1989a), encoding amino acids 1-76, fused to the N-terminus of the E-Prostanoid receptor, EP 2 . The human EP 2  open reading frame was inserted into a plasmid derivative of pRB200 between the EcoRI and XhoI sites by fusing it to the 3′ end of the sequence encoding the first 76 amino acids of lambda repressor using the EcoRI restriction enzyme site thereby creating pCK1-23. The stop codon of the human EP 2  receptor was removed from the hEP2 sequence by PCR. Briefly, the upstream oligonucleotide primer, originating from position 398 of the hEP 2  ORF, 5′-AGC GCT ACC TCT CGA TCG-3′ (SEQ ID NO:25), along with the downstream oligonucleotide primer, directed against the most 3′ region of hEP 2  ORF, 5′-GCC GCA  CTC GAG  GCA AGG TCA GCC TGT TTA CT-3′ (SEQ ID NO:26), were used in conjunction with VENT polymerase to amplify a new fragment lacking the stop codon from the hEP 2  ORF template (underlined sequence represents an XhoI site). Reaction conditions were carried out using 30 cycles of the following protocol: 1 min at 95° C., 15 sec at 98° C., 30 sec at 53° C. followed by 1 min at 72° C. The amplified product of 677 bp was digested at internal Bsu 36I and Xho I sites and the appropriate fragment subcloned into the plasmid pCK-1-23 to create the intermediate plasmid, pCK-1-38. Next, an oligonucleotide linker was synthesized to fuse the C-terminal portion of lambda repressor, in frame with the hEP 2  sequence. The sequence of this linker contains a thrombin cleavage site (aa sequence LVPRGS; SEQ ID NO:15), introduces a BamHI restriction site and begins with a 6× histidine tag. The two oligonucleotides, 5′-TCG AGC  CAC CAC CAC CAC CAC  TCT AGA CTG GTG CCA CGC G-3′ (SEQ ID NO:27) and 5′-GAT CCG CGT GGC ACC AGT CTA GA G TGG TGG TGG TGG TGG TG G C-3′ (SEQ ID NO:28), were annealed together at 65° C. and subcloned into pCK-1-38 following its digestion with Xho I and Bam HI, thereby creating the plasmid, CK2-5-HTL. The underlined sequences represent the histidine tag, while those in bold refer to the thrombin cleavage site.  
     [0084] Induction of cI-EP 2 -HTL fusion protein expression— E. coli  strain DH5α cells transformed with the pCK2-5-HTL expression plasmids were grown in 2×LB medium containing 100 μg/ml of ampicillin (2×LBA100). Cells were grown in 2×LBA100 with shaking at 37° C. until the culture reached an A 600  of 0.8. Protein expression was induced by addition of 1 mM isopropyl β-thiogalactoside (IPTG), followed by a further incubation at 30° C. for 5 hours. Then the cells were harvested by centrifugation at 2,500×g, flash frozen in liquid nitrogen and stored at −80° C.  
     [0085] Protein Purification by Ni-NTA Column—Bacterial cell pellets were resuspended in Buffer 1(50 nM Tris Cl, 150 mM Na Cl, 0.1% NaN3, 10 mM CHAPS, 20% glycerol, 2 mM PMSF, 1.4 mM β-ME, pH 8.0) followed by sonication three times for 20 sec, on ice. The protein was purified from inclusion bodies as described above for the cI-EP 3 -his protein.  
     [0086] Expression of cI-EP 2 -HTL fusion protein—The vector pCK2-5-HTL has the tac promoter and the cI translation initiator sequences. The cDNA encoding the human EP 2  receptor was modified as described above for the EP 3  receptor to obtain an EP 2  receptor fused to a portion of the cI gene (1-76 aa) at the N-terminus and a C-terminal fusion to a 6× his tag, followed by the thrombin cleavage site fused to amino acids 82-236 of λ-repressor. The resultant fusion protein of 610 aa has a predicted molecular weight 67.5 kDa. When expressed in  E. coli  this construct demonstrated high levels of expression of a protein of apparent molecular weight of 60 kDa in whole cell lysates when resolved by SDS-PAGE. The expression levels of the cI-EP 2 -HTL fusion protein were estimated to be in the range of 20 mg/L of  E. coli  culture. Western blot analysis of the lysates demonstrated that the induced protein of the appropriate molecular weight was reactive with the 24H antibody for EP 2  constructs tested.  
     [0087] Analysis of fusion of the EP 2  receptor to variable length cI leader sequences. To test the applicability of different bacterial promoters, while simultaneously assessing the limit to which the N-terminal sequence may be truncated, the human EP 2  receptor was fused to the bacteriophage T7 promoter with N-terminal fusion sequences of 0, 15, 36 and 76 aa of the N-terminus of cI. Induction of each of these fusion proteins was achieved by the addition of IPTG and the ability of each construct to express protein was assessed by Western blot. Initially, the N-terminal 76 aa of the cI gene were fused to the EP 2  receptor at an EcoRI restriction site encoding Glu-Phe at amino acids 75 and 76. The N-terminal sequence of 76 amino acids contains efficient translation initiation sequences and has 12 positively charged residues. To determine the minimum sequence requirement for efficient synthesis of recombinant protein from the cI fusion system, deletion analysis of the cI fusion sequence was performed. Deletions were performed by PCR mutagenesis employing an upstream primer which overlapped the initiator ATG and introduced an NdeI cloning site. The downstream primer overlapped the terminal sequence and introduced an EcoRI cloning site. These shortened N-terminal fusion sequences, consisting of amino acids 1-76 (SEQ ID NO:1; with 12 positively charged residues), amino acids 1-36 (SEQ ID NO:12; with 9 positively charged residues), amino acids 1-22 (SEQ ID NO:13; with 6 positively charged residues), amino acids 1-15 (SEQ ID NO:14; with 3 positively charged residues) and no leader (as control) were fused to the N-terminus of the EP 2  receptor and the resultant constructs were expressed in  E. coli . Protein expression was monitored by Western blot analysis, utilizing an anti-EP 2  sheep polyclonal antibody, as well as the Ni-NTA-HRP conjugate reactive with the 6×-his C-terminal fusion (FIG. 1). Because the 24H epitope lies within the first 36 aa of cI, this monoclonal anti -cI antibody was not useful for these experiments. In addition, these constructs were expressed in the pT7 “pET” vectors (Novagen, Madison, Wis. to compare the efficacy of the T7 promoter versus the previously described ptac promoter. The T7 promoter has the advantage of being less “leaky” than tac, i.e., it has a lower basal level of transcription. The results of these comparative studies demonstrated that the ptac promoter was superior to the T7 promoter in yielding high steady state levels of protein. Overall, the greatest amount of expression was observed with the cI 1-76  construct, no expression was observed with the cI 0  (no leader sequence) construct and intermediate amounts of expression were observed with the cI 1-15 , cI 1-22  and cI 1-36  constructs.  
     Example 3  
     Overexpression of Non EP Receptor Proteins  
     [0088] The ability of the ci fusion system of this invention to produce a variety of proteins was assessed as follows: the pLJM5-22H plasmid encoding the fusion of the PGEP 2 EP 3  receptor with the N-terminal domain of λ repressor was modified to remove the cDNA encoding the PGEP 2 EP 3  receptor and DNA sequences encoding alternative target proteins were inserted as fusion proteins in-frame with and downstream from the tac promoter/cI fusion at amino acid 76 of the λ repressor protein. As a general plasmid construction strategy, the target protein was amplified by polymerase chain reaction using an upstream oligonucleotide which overlapped the N-terminal sequence of the target protein and introduced an NdeI site immediately upstream from the ATG start codon. The downstream oligonucleotide overlapped the final codons of the target protein (in general, six codons), removed the stop codon and introduced an XhoI site. This allowed introduction of the target sequence at the NdeI and XhoI restriction sites which flanked the PGEP 2 EP 3  receptor in plasmid pLJM5-22H. The resulting constructs were fused at the N-terminus to the first 76 aa of λ repressor and at the C-terminus to the 6×-his tag.  
     [0089] Plasmid pSD1.63 his-fusion of the human CCR5 receptor with the N-terminal domain of A repressor. Construct pSD1.63 his (SEQ ID NO:3) was assembled according to the general outline as described above. The plasmid allowed expression of the nucleotide sequence encoding the CCR5 protein, a chemokine receptor of the GPCR superfamily and the co-receptor for HIV entry into host cells.  
     [0090] The upstream oligonucleotide primer had the sequence: 5′ GCGC  CAT ATG  GAT TAT AAG TGT CAA GTC CAA 3′ (SEQ ID NO:29). The downstream primer had the sequence: 5′ GCCG  CT CGA G GC CAA GCC CAC AGA TAT TTC CT 3′ (SEQ ID NO:30).  
     [0091] The underlined sequence delineates the NdeI sequence in the upper primer and the XhoI sequence in the lower primer, respectively. The two primers were used to amplify the CCR5 receptor from human genomic DNA utilizing the following reaction conditions: 35 cycles of the following protocol: 1 min at 95° C., 15 sec at 95° C., 15 sec at 68° C., followed by a 10 min final extension at 72° C. The resultant amplified product of 1074 nt was digested with NdeI and XhoI and inserted into the expression vector.  
     [0092] The fusion protein was produced and purified as described for the PGEP 2 EP 3  receptor, with modifications for optimal growth, induction times and temperatures. In brief, the culture conditions were as follows: Medium was inoculated with a 1:100 dilution of a fresh overnight culture and grown overnight for 24 hours at 30° C. in 2×LB medium containing 2% glucose and the appropriate antibiotics. Cells were collected by centrifugation and resuspended in 2×LB (no glucose) containing 1 mM IPTG and the appropriate antibiotics. Cells were then grown for varying times of induction at 30° C. and harvested by centrifugation. For the CCR-5 fusion protein, induction was overnight for 24 hours.  
     [0093] Plasmid pSD1.18 his-fusion of the human β 2 AR receptor with the N-terminal domain of λ repressor. Construct pSD1.18 his (SEQ ID NO:4) was assembled using a modification of the general outline described above. The natural stop codon of the semi-synthetic β 2 AR was removed by PCR, however the 5′ end of the cDNA was subcloned into the EcoRI site at codon 75/76 of λ repressor, which is immediately upstream of the flanking NdeI site utilized in the previous constructs.  
     [0094] The sense oligonucleotide contains sequence internal to the β 2 AR and adds an Nco I and EcoRI site at the 5′ end: 5′ GCGC GAATTC A CCATG G AA ATG AGA CCT GCT GTG ACT TC 3′ (SEQ ID NO:31).  
     [0095] The mutagenic antisense oligonucleotide primer which removes the stop codon and introduces the XhoI site for fusion to the 6× his tag had the sequence: 5′CCGGG  CT CGA G GC TAG CAGTGA GTC ATT TGT ACT ACA AT 3′ (SEQ ID NO:32).  
     [0096] The underlined sequence delineates the EcoRI and NcoI restriction sequences in the upper sense primer and the XhoI sequence in the lower antisense primer. These oligonucleotides were used to amplify a small internal β 2 AR sequence with convenient EcoRI and XhoI restriction sites at the termini. This fragment was subcloned into the tac/cI fusion expression vector as an intermediate step. The resulting plasmid was digested with at NcoI and an (internal to β 2 AR) EcoRV site contained within the original PCR fragment. The corresponding NcoI-EcoRV fragment of the β 2 AR cDNA was subcloned into this site, reconstituting the fill length modified β 2 AR sequence. This resulted in the cI 1-76 -β 2 AR-6× his fusion protein cloned into the cI-fusion expression vector. The fusion protein was produced and purified as described herein for the EP 3  receptor with an overnight 24 hour induction.  
     [0097] Plasmid pSD 1.134 his-fusion of the ROMK receptor with the N-terminal domain of λ repressor. Construct pSD1.134 his (SEQ ID NO:5) was assembled according to the general outline as described herein. The upstream oligonucleotide primer had the sequence: 5′ GG GAATTC CAT ATG  TTC AAA CAC CTC CGA AGA TGG 3′ (SEQ ID NO:33). The downstream primer had the sequence: 5′ CCG CTCGAG GC CAT CTG GGT GTC GTC CGT TTCA TC 3′ (SEQ ID NO:34). The underlined sequence delineates EcoRI and NdeI sequences in the upper primer and the XhoI sequence in the lower primer, respectively. The two primers were used to amplify the ROMK channel from the cloned rat cDNA utilizing PCR. The resultant amplified product was digested with NdeI and XhoI and the 1.1 kb fragment was inserted into the expression vector. The resultant cI 1-76  fusion protein was under the control of the tac promoter. The fusion protein was produced and purified as described herein for the PGEP 2 EP 3  receptor, with a three to five hour induction. The resulting yield of ROMK protein by this method was approximately 1 mg of purified protein/liter of bacterial culture when induced at A 600 =1 to 2.  
     [0098] Plasmid pSD2.46his-fusion of the human rho protein with the N-terminal domain of λ repressor. Construct pSD2.46his (SEQ ID NO:6) was assembled according to the general outline described herein. The resulting construct allowed expression of the nucleotide sequence encoding a fusion protein of cI 1-76  and the human rho protein, which is a cytoplasmic protein which is membrane-associated and a member of the family of small G-proteins. The upstream oligonucleotide primer had the sequence: 5′ GCGCG C ATATG GCTGCCATCCGGAAG3′ (SEQ ID NO:35). The downstream primer had the sequence: 5′ GCC G CT CGA G GC CAA GAC AAG GCA ACC AGA3′ (SEQ ID NO:36). The underlined sequence delineates an NdeI sequence in the upper primer and an XhoI sequence in the lower primer, respectively. The two primers were used to amplify rho from cloned human cDNA utilizing PCR. The resultant amplified product was digested with NdeI and XhoI and the 0.6 kb fragment was inserted into the expression vector. The resultant cI 1-76  fusion protein was under the control of the tac promoter. The fusion protein was produced and purified as described herein for the PGEP 2 EP 3  receptor, with a three to 24 hour induction.  
     [0099] Although the present process has been described with reference to specific details of certain embodiments thereof, it is not intended that such details should be regarded as limitations upon the scope of the invention except as and to the extent that they are included in the accompanying claims.  
     [0100] Throughout this application, various publications are referenced. The disclosures of these publications in their entireties are hereby incorporated by reference into this application in order to more fully describe the state of the art to which this invention pertains.  
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     [0112] Limbird, L. E.  Cell surface receptors: A short course on theory and methods . Second edition. Kluwer Academic Publishers, Norwell, Mass. (1996).  
     [0113] Loisel, T. P., H. Ansanay, S. St-Onge, B. Gay, P. Boulanger, A. D. Strosberg, S. Marullo and M. Bouvier. Recovery of homogeneous and functional β 2  adrenergic receptors from extracellular baculovirus particles.  Nature Biotechnology  15:1300-1304 (1997).  
     [0114] Marullo, S., C. Delavier-Klutchko, Y. Eshdat, A. D. Strosberg and L. J. Emorine. Human β 2  adrenergic receptors expressed in  E. coli  membranes retain their pharmacological properties.  Proc. Natl. Acad. Sci. U.S.A.  85(October 1988):7551-7555 (1988).  
     [0115] Michieli, et al.  Oncogene  12:775-784 (1996).  
     [0116] Sambrook et al.  Molecular Cloning: A Laboratory Manual.  2d Ed. Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y. 1988.  
     [0117] Sander, P., S. Grünewald, Reilander and H. Michel. Expression of the human D2s dopamine receptor in yeasts  Saccharomyces cerevisae  and  Schizosaccharomyces pombe : a comparative study.  FEBS Letters  344:41-46 (1994).  
     [0118] Sauer, R. T.  Biochemistry  17:1092-1100 (1978).  
     [0119] Sauer et al.  Nature  279:396-400 (1979).  
     [0120] Zeidel, M. L., et al.  Biochemistry  31:7436-7441 (1992).  
     [0121] Zweichal, A., et al.  Am. J. Physiol.  261:F187-F196 (1991).  
     [0122] 
    
     
       
         1 
         
           
             37  
           
           
             1  
             5856  
             DNA  
             Artificial Sequence  
             
               CDS  
               (300)...(1799)  
               Description of Artificial Sequence/Note =
      synthetic construct  
             
           
            1 

ttctcatgtt tgacagctta tctcatcgac tgcacggtgc accaatgctt ctggcgtcag     60 

gcagccatcg gaagctgtgg tatggctgtg caggtcgtaa atcactgcat aattcgtgtc    120 

gctcaaggcg cactcccgtt ctggataatg ttttttgcgc cgacatcata acggttctgg    180 

caaatattct gaaatgagct gttgacaatt aatcatcggc tcgtataatg tggaattgtg    240 

agcggataac aattaatgtg tgaatgtgag cggatacaat ttcacacagg aaacagcgt     299 

atg agc aca aaa aag aaa cca tta aca caa gag cag ctt gag gac gca      347 
Met Ser Thr Lys Lys Lys Pro Leu Thr Gln Glu Gln Leu Glu Asp Ala 
 1               5                   10                  15 

cgt cgc ctt aaa gca att tat gaa aaa aag aaa aat gaa ctt ggc tta      395 
Arg Arg Leu Lys Ala Ile Tyr Glu Lys Lys Lys Asn Glu Leu Gly Leu 
             20                  25                  30 

tcc cag gaa tct gtc gca gac aag atg ggg atg ggg cag tca ggc gtt      443 
Ser Gln Glu Ser Val Ala Asp Lys Met Gly Met Gly Gln Ser Gly Val 
         35                  40                  45 

ggt gct tta ttt aat ggc atc aat gca tta aat gct tat aac gcg gca      491 
Gly Ala Leu Phe Asn Gly Ile Asn Ala Leu Asn Ala Tyr Asn Ala Ala 
     50                  55                  60 

ttg cta gca aaa att ctc aaa gtt agc gtt gaa gaa ttc gca gct cat      539 
Leu Leu Ala Lys Ile Leu Lys Val Ser Val Glu Glu Phe Ala Ala His 
 65                  70                  75                  80 

atg aag gag acg cgg ggc gac gga ggg agc gcc ccc ttc tgc acc cgc      587 
Met Lys Glu Thr Arg Gly Asp Gly Gly Ser Ala Pro Phe Cys Thr Arg 
                 85                  90                  95 

ctc aac cac tcg tat cca ggc atg tgg gcg ccc gag gca cgg ggc aac      635 
Leu Asn His Ser Tyr Pro Gly Met Trp Ala Pro Glu Ala Arg Gly Asn 
            100                 105                 110 

ctc aca cgc ccc cca ggg ccc ggc gag gac tgt ggc tcg gtg tcc gtg      683 
Leu Thr Arg Pro Pro Gly Pro Gly Glu Asp Cys Gly Ser Val Ser Val 
        115                 120                 125 

gcc ttc ccg atc acc atg ctg atc acc ggc ttc gtg ggc aac gcg ctg      731 
Ala Phe Pro Ile Thr Met Leu Ile Thr Gly Phe Val Gly Asn Ala Leu 
    130                 135                 140 

gcc atg ctg ctc gtg tcg cgt agc tac cgg cgt cgg gag agc aag cgc      779 
Ala Met Leu Leu Val Ser Arg Ser Tyr Arg Arg Arg Glu Ser Lys Arg 
145                 150                 155                 160 

aag aag tcg ttc ctg ttg tgc atc ggc tgg ctg gcg ctc act gac ctg      827 
Lys Lys Ser Phe Leu Leu Cys Ile Gly Trp Leu Ala Leu Thr Asp Leu 
                165                 170                 175 

gtc ggg cag ctg ctc aca agc ccc gtg gtc atc ttg gtg tac cta tcc      875 
Val Gly Gln Leu Leu Thr Ser Pro Val Val Ile Leu Val Tyr Leu Ser 
            180                 185                 190 

aag cag cgc tgg gag cag ctc gac ccg tcg ggg cgc ctg tgc acc ttc      923 
Lys Gln Arg Trp Glu Gln Leu Asp Pro Ser Gly Arg Leu Cys Thr Phe 
        195                 200                 205 

ttt ggt ctg acc atg act gtt ttc ggg ctg tcc tcg ctc ttc atc gcc      971 
Phe Gly Leu Thr Met Thr Val Phe Gly Leu Ser Ser Leu Phe Ile Ala 
    210                 215                 220 

agc gcc atg gct gtc gag agg gcg ctg gcc atc cgt gcg cca cac tgg     1019 
Ser Ala Met Ala Val Glu Arg Ala Leu Ala Ile Arg Ala Pro His Trp 
225                 230                 235                 240 

tac gcg agc cac atg aag acg cgt gcc act cgc gcc gtc ctg ctg ggc     1067 
Tyr Ala Ser His Met Lys Thr Arg Ala Thr Arg Ala Val Leu Leu Gly 
                245                 250                 255 

gtg tgg ctg gca gtg ctc gcc ttc gcc ctg cta cct gtg ctg ggt gtg     1115 
Val Trp Leu Ala Val Leu Ala Phe Ala Leu Leu Pro Val Leu Gly Val 
            260                 265                 270 

ggt cag tac acc atc cag tgg ccc ggg acg tgg tgc ttc atc agc acc     1163 
Gly Gln Tyr Thr Ile Gln Trp Pro Gly Thr Trp Cys Phe Ile Ser Thr 
        275                 280                 285 

gga cga ggg gac aac ggg acg agc tct tca cac aac tgg ggc aac ctt     1211 
Gly Arg Gly Asp Asn Gly Thr Ser Ser Ser His Asn Trp Gly Asn Leu 
    290                 295                 300 

ttc ttc gcc tcc acc ttt gcc ttc ctg ggc ctc ttg gcg ctg gcc atc     1259 
Phe Phe Ala Ser Thr Phe Ala Phe Leu Gly Leu Leu Ala Leu Ala Ile 
305                 310                 315                 320 

acc ttc acc tgc aac ctg gcc acc att aag gct ctg gtg tcc cgc tgc     1307 
Thr Phe Thr Cys Asn Leu Ala Thr Ile Lys Ala Leu Val Ser Arg Cys 
                325                 330                 335 

cgg gca aag gcg gca gca tca cag tcc agt gcc cag tgg ggc cgg atc     1355 
Arg Ala Lys Ala Ala Ala Ser Gln Ser Ser Ala Gln Trp Gly Arg Ile 
            340                 345                 350 

acg acc gag acg gcc atc cag ctc atg ggg atc atg tgc gtg ctg tcg     1403 
Thr Thr Glu Thr Ala Ile Gln Leu Met Gly Ile Met Cys Val Leu Ser 
        355                 360                 365 

gtc tgc tgg tcg ccc cta ctg ata atg atg ttg aaa atg atc ttc aat     1451 
Val Cys Trp Ser Pro Leu Leu Ile Met Met Leu Lys Met Ile Phe Asn 
    370                 375                 380 

cag aca tca gtt gag cac tgc aag aca gac aca gga aag cag aaa gaa     1499 
Gln Thr Ser Val Glu His Cys Lys Thr Asp Thr Gly Lys Gln Lys Glu 
385                 390                 395                 400 

tgc aac ttc ttc tta ata gct gtt cgc ctg gct tca ctg aac cag ata     1547 
Cys Asn Phe Phe Leu Ile Ala Val Arg Leu Ala Ser Leu Asn Gln Ile 
                405                 410                 415 

ttg gat ccc tgg gtt tat ctg ctg cta aga aag att ctt ctt cgg aag     1595 
Leu Asp Pro Trp Val Tyr Leu Leu Leu Arg Lys Ile Leu Leu Arg Lys 
            420                 425                 430 

ttt tgc cag gta att cat gaa aat aat gag cag aag gat gaa att cag     1643 
Phe Cys Gln Val Ile His Glu Asn Asn Glu Gln Lys Asp Glu Ile Gln 
        435                 440                 445 

cgt gag aac agg aac gtc tca cac agt ggg caa cac gaa gag gcc aga     1691 
Arg Glu Asn Arg Asn Val Ser His Ser Gly Gln His Glu Glu Ala Arg 
    450                 455                 460 

gac agt gag aag agc aaa acc atc cct ggc ctg ttc tcc att ctg ctg     1739 
Asp Ser Glu Lys Ser Lys Thr Ile Pro Gly Leu Phe Ser Ile Leu Leu 
465                 470                 475                 480 

cag gct gac cct ggt gct cgt cct tat cag caa gcc tcg agg cac cat     1787 
Gln Ala Asp Pro Gly Ala Arg Pro Tyr Gln Gln Ala Ser Arg His His 
                485                 490                 495 

cac cac cac cac tgaagcttta atgcggtagt ttatcacagt taaattgcta         1839 
His His His His 
            500 

acgcagtcag gcaccgtgta tgaaatctaa caatgcgctc atcgtcatcc tcggcaccgt   1899 

caccctggat gctgtaggca taggcttggt tatgccggta ctgccgggcc tcttgcggga   1959 

tcgacgcgag gctggatggc cttccccatt atgattcttc tcgcttccgg cggcatcggg   2019 

atgcccgcgt tgcaggccat gctgtccagg caggtagatg acgaccatca gggacagctt   2079 

caaggatcgc tcgcggctct taccagccta acttcgatca ctggaccgct gatcgtcacg   2139 

gcgatttatg ccgcctcggc gagcacatgg aacgggttgg catggattgt aggcgccgcc   2199 

ctataccttg tctgcctccc cgcgttgcgt cgcggtgcat ggagccgggc cacctcgacc   2259 

tgaatggaag ccggcggcac ctcgctaacg gattcaccac tccaagaatt ggagccaatc   2319 

aattcttgcg gagaactgtg aatgcgcaaa ccaacccttg gcagaacata tccatcgcgt   2379 

ccgccatctc cagcagccgc acgcggcgca tctcgggcag cgttgggtcc tggccacggg   2439 

tgcgcatgat cgtgctcctg tcgttgagga cccggctagg ctggcggggt tgccttactg   2499 

gttagcagaa tgaatcaccg atacgcgagc gaacgtgaag cgactgctgc tgcaaaacgt   2559 

ctgcgacctg agcaacaaca tgaatggtct tcggtttccg tgtttcgtaa agtctggaaa   2619 

cgcggaagtc agcgccctgc accattatgt tccggatctg catcgcagga tgctgctggc   2679 

taccctgtgg aacacctaca tctgtattaa cgaagcgctg gcattgaccc tgagtgattt   2739 

ttctctggtc ccgccgcatc cataccgcca gttgtttacc ctcacaacgt tccagtaacc   2799 

gggcatgttc atcatcagta acccgtatcg tgagcatcct ctctcgtttc atcggtatca   2859 

ttacccccat gaacagaaat tcccccttac acggaggcat caagtgacca aacaggaaaa   2919 

aaccgccctt aacatggccc gctttatcag aagccagaca ttaacgcttc tggagaaact   2979 

caacgagctg gacgcggatg aacaggcaga catctgtgaa tcgcttcacg accacgctga   3039 

tgagctttac cgcagctgcc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat   3099 

gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg   3159 

tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc agtcacgtag   3219 

cgatagcgga gtgtatactg gcttaactat gcggcatcag agcagattgt actgagagtg   3279 

caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc   3339 

tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta   3399 

tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag   3459 

aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg   3519 

tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg   3579 

tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg   3639 

cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga   3699 

agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc   3759 

tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt   3819 

aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact   3879 

ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg   3939 

cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt   3999 

accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt   4059 

ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct   4119 

ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg   4179 

gtcatgagat tatcaaaaag gatcttcacc tagatccttt taccccggtt gataatcaga   4239 

aaagccccaa aaacaggaag attgtataag caaatattta aattgtaaac gttaatattt   4299 

tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa   4359 

tcggcaaaat cccttataaa tcaaaagaat agcccgagat agggttgagt gttgttccag   4419 

tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg   4479 

tctatcaggg cgatggccca ctacgtgaac catcacccaa atcaagtttt ttggggtcga   4539 

ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg   4599 

gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg   4659 

cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc   4719 

cgctacaggg cgcgtaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac   4779 

caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt   4839 

gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt   4899 

gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag   4959 

ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct   5019 

attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt   5079 

gttgccattg ctgcaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc   5139 

tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt   5199 

agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg   5259 

gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg   5319 

actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct   5379 

tgcccggcgt caacacggga taataccgcg ccacatagca gaactttaaa agtgctcatc   5439 

attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt   5499 

tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt   5559 

tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg   5619 

aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat   5679 

tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg   5739 

cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta   5799 

acctataaaa ataggcgtat cacgaggccc tttcgtcttc aagaattgat cgatcaa      5856 

 
           
             2  
             6446  
             DNA  
             Artificial Sequence  
             
               CDS  
               (300)...(2126)  
               Description of Artificial Sequence/note =
      synthetic construct  
             
           
            2 

ttctcatgtt tgacagctta tctcatcgac tgcacggtgc accaatgctt ctggcgtcag     60 

gcagccatcg gaagctgtgg tatggctgtg caggtcgtaa atcactgcat aattcgtgtc    120 

gctcaaggcg cactcccgtt ctggataatg ttttttgcgc cgacatcata acggttctgg    180 

caaatattct gaaatgagct gttgacaatt aatcatcggc tcgtataatg tggaattgtg    240 

agcggataac aattaatgtg tgaatgtgag cggatacaat ttcacacagg aaacagcgt     299 

atg agc aca aaa aag aaa cca tta aca caa gag cag ctt gag gac gca      347 
Met Ser Thr Lys Lys Lys Pro Leu Thr Gln Glu Gln Leu Glu Asp Ala 
 1               5                   10                  15 

cgt cgc ctt aaa gca att tat gaa aaa aag aaa aat gaa ctt ggc tta      395 
Arg Arg Leu Lys Ala Ile Tyr Glu Lys Lys Lys Asn Glu Leu Gly Leu 
             20                  25                  30 

tcc cag gaa tct gtc gca gac aag atg ggg atg ggg cag tca ggc gtt      443 
Ser Gln Glu Ser Val Ala Asp Lys Met Gly Met Gly Gln Ser Gly Val 
         35                  40                  45 

ggt gct tta ttt aat ggc atc aat gca tta aat gct tat aac gcg gca      491 
Gly Ala Leu Phe Asn Gly Ile Asn Ala Leu Asn Ala Tyr Asn Ala Ala 
     50                  55                  60 

ttg cta gca aaa att ctc aaa gtt agc gtt gaa gaa ttc cat atg ggc      539 
Leu Leu Ala Lys Ile Leu Lys Val Ser Val Glu Glu Phe His Met Gly 
 65                  70                  75                  80 

aat gcc tcc aat gac tcc cag tct gag gac tgc gag acg cga cag tgg      587 
Asn Ala Ser Asn Asp Ser Gln Ser Glu Asp Cys Glu Thr Arg Gln Trp 
                 85                  90                  95 

ctt ccc cca ggc gaa agc cca gcc atc agc tcc gtc atg ttc tcg gcc      635 
Leu Pro Pro Gly Glu Ser Pro Ala Ile Ser Ser Val Met Phe Ser Ala 
            100                 105                 110 

ggg gtg ctg ggg aac ctc ata gca ctg gcg ctg ctg gcg cgc cgc tgg      683 
Gly Val Leu Gly Asn Leu Ile Ala Leu Ala Leu Leu Ala Arg Arg Trp 
        115                 120                 125 

cgg ggg gac gtg ggg tgc agc gcc ggc cgc agg agc tcc ctc tcc ttg      731 
Arg Gly Asp Val Gly Cys Ser Ala Gly Arg Arg Ser Ser Leu Ser Leu 
    130                 135                 140 

ttc cac gtg ctg gtg acc gag ctg gtg ttc acc gac ctg ctc ggg acc      779 
Phe His Val Leu Val Thr Glu Leu Val Phe Thr Asp Leu Leu Gly Thr 
145                 150                 155                 160 

tgc ctc atc agc cca gtg gta ctg gct tcg tac gcg cgg aac cag acc      827 
Cys Leu Ile Ser Pro Val Val Leu Ala Ser Tyr Ala Arg Asn Gln Thr 
                165                 170                 175 

ctg gtg gca ctg gcg ccc gag agc cgc gcg tgc acc tac ttc gct ttc      875 
Leu Val Ala Leu Ala Pro Glu Ser Arg Ala Cys Thr Tyr Phe Ala Phe 
            180                 185                 190 

gcc atg acc ttc ttc agc ctg gcc acg atg ctc atg ctc ttc gcc atg      923 
Ala Met Thr Phe Phe Ser Leu Ala Thr Met Leu Met Leu Phe Ala Met 
        195                 200                 205 

gcc ctg gag cgc tac ctc tcg atc ggg cac ccc tac ttc tac cag cgc      971 
Ala Leu Glu Arg Tyr Leu Ser Ile Gly His Pro Tyr Phe Tyr Gln Arg 
    210                 215                 220 

cgc gtc tcg gcc tcc ggg ggc ctg gcc gtg ctg cct gtc atc tat gca     1019 
Arg Val Ser Ala Ser Gly Gly Leu Ala Val Leu Pro Val Ile Tyr Ala 
225                 230                 235                 240 

gtc tcc ctg ctc ttc tgc tcg ctg ccg ctg ctg gac tat ggg cag tac     1067 
Val Ser Leu Leu Phe Cys Ser Leu Pro Leu Leu Asp Tyr Gly Gln Tyr 
                245                 250                 255 

gtc cag tac tgc ccc ggg acc tgg tgc ttc atc cgg cac ggg cgg acc     1115 
Val Gln Tyr Cys Pro Gly Thr Trp Cys Phe Ile Arg His Gly Arg Thr 
            260                 265                 270 

gct tac ctg cag ctg tac gcc acc ctg ctg ctg ctt ctc att gtc tcg     1163 
Ala Tyr Leu Gln Leu Tyr Ala Thr Leu Leu Leu Leu Leu Ile Val Ser 
        275                 280                 285 

gtg ctc gcc tgc aac ttc agt gtc att ctc aac ctc atc cgc atg cac     1211 
Val Leu Ala Cys Asn Phe Ser Val Ile Leu Asn Leu Ile Arg Met His 
    290                 295                 300 

cgc cga agc cgg aga agc cgc tgc gga cct tcc ctg ggc agt ggc cgg     1259 
Arg Arg Ser Arg Arg Ser Arg Cys Gly Pro Ser Leu Gly Ser Gly Arg 
305                 310                 315                 320 

ggc ggc ccc ggg gcc cgc agg aga ggg gaa agg gtg tcc atg gcg gag     1307 
Gly Gly Pro Gly Ala Arg Arg Arg Gly Glu Arg Val Ser Met Ala Glu 
                325                 330                 335 

gag acg gac cac ctc att ctc ctg gct atc atg acc atc acc ttc gcc     1355 
Glu Thr Asp His Leu Ile Leu Leu Ala Ile Met Thr Ile Thr Phe Ala 
            340                 345                 350 

gtc tgc tcc ttg cct ttc acg att ttt gca tat atg aat gaa acc tct     1403 
Val Cys Ser Leu Pro Phe Thr Ile Phe Ala Tyr Met Asn Glu Thr Ser 
        355                 360                 365 

tcc cga aag gaa aaa tgg gac ctc caa gct ctt agg ttt tta tca att     1451 
Ser Arg Lys Glu Lys Trp Asp Leu Gln Ala Leu Arg Phe Leu Ser Ile 
    370                 375                 380 

aat tca ata att gac cct tgg gtc ttt gcc atc ctt agg cct cct gtt     1499 
Asn Ser Ile Ile Asp Pro Trp Val Phe Ala Ile Leu Arg Pro Pro Val 
385                 390                 395                 400 

ctg aga cta atg cgt tca gtc ctc tgt tgt cgg att tca tta aga aca     1547 
Leu Arg Leu Met Arg Ser Val Leu Cys Cys Arg Ile Ser Leu Arg Thr 
                405                 410                 415 

caa gat gca aca caa act tcc tgt tct aca cag tca gat gcc agt aaa     1595 
Gln Asp Ala Thr Gln Thr Ser Cys Ser Thr Gln Ser Asp Ala Ser Lys 
            420                 425                 430 

cag gct gac ctt gcc tcg agc cac cac cac cac cac cac tct aga ctg     1643 
Gln Ala Asp Leu Ala Ser Ser His His His His His His Ser Arg Leu 
        435                 440                 445 

gtg cca cgc gga tcc gtt cga gaa atc tac gag atg tat gaa gcg gtt     1691 
Val Pro Arg Gly Ser Val Arg Glu Ile Tyr Glu Met Tyr Glu Ala Val 
    450                 455                 460 

agc atg cag ccg tca ctt aga agt gag tat gag tac cct gtt ttt tct     1739 
Ser Met Gln Pro Ser Leu Arg Ser Glu Tyr Glu Tyr Pro Val Phe Ser 
465                 470                 475                 480 

cat gtt cag gca ggg atg ttc tca cct aag ctt aga acc ttt acc aaa     1787 
His Val Gln Ala Gly Met Phe Ser Pro Lys Leu Arg Thr Phe Thr Lys 
                485                 490                 495 

ggt gat gcg gag aga tgg gta agc aca acc aaa aaa gcc agt gat tct     1835 
Gly Asp Ala Glu Arg Trp Val Ser Thr Thr Lys Lys Ala Ser Asp Ser 
            500                 505                 510 

gca ttc tgg ctt gag gtt gaa ggt aat tcc atg acc gca cca aca ggc     1883 
Ala Phe Trp Leu Glu Val Glu Gly Asn Ser Met Thr Ala Pro Thr Gly 
        515                 520                 525 

tcc aag cca agc ttt cct gac gga atg tta att ctc gtt gac cct gag     1931 
Ser Lys Pro Ser Phe Pro Asp Gly Met Leu Ile Leu Val Asp Pro Glu 
    530                 535                 540 

cag gct gtt gag cca ggt gat ttc tgc ata gcc aga ctt ggg ggt gat     1979 
Gln Ala Val Glu Pro Gly Asp Phe Cys Ile Ala Arg Leu Gly Gly Asp 
545                 550                 555                 560 

gag ttt acc ttc aag aaa ctg atc agg gat agc ggt cag gtg ttt tta     2027 
Glu Phe Thr Phe Lys Lys Leu Ile Arg Asp Ser Gly Gln Val Phe Leu 
                565                 570                 575 

caa cca cta aac cca cag tac cca atg atc cca tgc aat gag agt tgt     2075 
Gln Pro Leu Asn Pro Gln Tyr Pro Met Ile Pro Cys Asn Glu Ser Cys 
            580                 585                 590 

tcc gtt gtg ggg aaa gtt atc gct agt cag tgg cct gaa gag acg ttt     2123 
Ser Val Val Gly Lys Val Ile Ala Ser Gln Trp Pro Glu Glu Thr Phe 
        595                 600                 605 

ggc tgatcggcaa ggtgttctgg tcggcgcata gctgataaca attgagcaag          2176 
Gly 

aatcttcatc gaattagggg aattttcact cccctcagaa cataacatag taaatggatt   2236 

gaattatgaa gaatggtttt tatgcgactt accgcagcaa aaataaaggg aaagataagc   2296 

gctcaataaa cctgtctgtt ttccttaatt ctctgctggc tgataatcat cacctgcagg   2356 

ttggctccaa ttatttgtat attcataaaa tcgataagct ttaatgcggt agtttatcac   2416 

agttaaattg ctaacgcagt caggcaccgt gtatgaaatc taacaatgcg ctcatcgtca   2476 

tcctcggcac cgtcaccctg gatgctgtag gcataggctt ggttatgccg gtactgccgg   2536 

gcctcttgcg ggatcgacgc gaggctggat ggccttcccc attatgattc ttctcgcttc   2596 

cggcggcatc gggatgcccg cgttgcaggc catgctgtcc aggcaggtag atgacgacca   2656 

tcagggacag cttcaaggat cgctcgcggc tcttaccagc ctaacttcga tcactggacc   2716 

gctgatcgtc acggcgattt atgccgcctc ggcgagcaca tggaacgggt tggcatggat   2776 

tgtaggcgcc gccctatacc ttgtctgcct ccccgcgttg cgtcgcggtg catggagccg   2836 

ggccacctcg acctgaatgg aagccggcgg cacctcgcta acggattcac cactccaaga   2896 

attggagcca atcaattctt gcggagaact gtgaatgcgc aaaccaaccc ttggcagaac   2956 

atatccatcg cgtccgccat ctccagcagc cgcacgcggc gcatctcggg cagcgttggg   3016 

tcctggccac gggtgcgcat gatcgtgctc ctgtcgttga ggacccggct aggctggcgg   3076 

ggttgcctta ctggttagca gaatgaatca ccgatacgcg agcgaacgtg aagcgactgc   3136 

tgctgcaaaa cgtctgcgac ctgagcaaca acatgaatgg tcttcggttt ccgtgtttcg   3196 

taaagtctgg aaacgcggaa gtcagcgccc tgcaccatta tgttccggat ctgcatcgca   3256 

ggatgctgct ggctaccctg tggaacacct acatctgtat taacgaagcg ctggcattga   3316 

ccctgagtga tttttctctg gtcccgccgc atccataccg ccagttgttt accctcacaa   3376 

cgttccagta accgggcatg ttcatcatca gtaacccgta tcgtgagcat cctctctcgt   3436 

ttcatcggta tcattacccc catgaacaga aattccccct tacacggagg catcaagtga   3496 

ccaaacagga aaaaaccgcc cttaacatgg cccgctttat cagaagccag acattaacgc   3556 

ttctggagaa actcaacgag ctggacgcgg atgaacaggc agacatctgt gaatcgcttc   3616 

acgaccacgc tgatgagctt taccgcagct gcctcgcgcg tttcggtgat gacggtgaaa   3676 

acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga   3736 

gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc gcagccatga   3796 

cccagtcacg tagcgatagc ggagtgtata ctggcttaac tatgcggcat cagagcagat   3856 

tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata   3916 

ccgcatcagg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct   3976 

gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga   4036 

taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc   4096 

cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg   4156 

ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg   4216 

aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt   4276 

tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt   4336 

gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg   4396 

cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact   4456 

ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt   4516 

cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct   4576 

gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac   4636 

cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc   4696 

tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg   4756 

ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaccccg   4816 

gttgataatc agaaaagccc caaaaacagg aagattgtat aagcaaatat ttaaattgta   4876 

aacgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac   4936 

caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagcccga gatagggttg   4996 

agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa   5056 

gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc caaatcaagt   5116 

tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt   5176 

agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga   5236 

gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc   5296 

gcgcttaatg cgccgctaca gggcgcgtaa atcaatctaa agtatatatg agtaaacttg   5356 

gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg   5416 

ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc   5476 

atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc   5536 

agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc   5596 

ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag   5656 

tttgcgcaac gttgttgcca ttgctgcagg catcgtggtg tcacgctcgt cgtttggtat   5716 

ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg   5776 

caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt   5836 

gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag   5896 

atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg   5956 

accgagttgc tcttgcccgg cgtcaacacg ggataatacc gcgccacata gcagaacttt   6016 

aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct   6076 

gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac   6136 

tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat   6196 

aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat   6256 

ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca   6316 

aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat   6376 

tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc ttcaagaatt   6436 

gatcgatcaa                                                          6446 

 
           
             3  
             5674  
             DNA  
             Artificial Sequence  
             
               CDS  
               (300)...(1616)  
               Description of Artificial Sequence/note =
      synthetic construct  
             
           
            3 

ttctcatgtt tgacagctta tctcatcgac tgcacggtgc accaatgctt ctggcgtcag     60 

gcagccatcg gaagctgtgg tatggctgtg caggtcgtaa atcactgcat aattcgtgtc    120 

gctcaaggcg cactcccgtt ctggataatg ttttttgcgc cgacatcata acggttctgg    180 

caaatattct gaaatgagct gttgacaatt aatcatcggc tcgtataatg tggaattgtg    240 

agcggataac aattaatgtg tgaatgtgag cggatacaat ttcacacagg aaacagcgt     299 

atg agc aca aaa aag aaa cca tta aca caa gag cag ctt gag gac gca      347 
Met Ser Thr Lys Lys Lys Pro Leu Thr Gln Glu Gln Leu Glu Asp Ala 
 1               5                   10                  15 

cgt cgc ctt aaa gca att tat gaa aaa aag aaa aat gaa ctt ggc tta      395 
Arg Arg Leu Lys Ala Ile Tyr Glu Lys Lys Lys Asn Glu Leu Gly Leu 
             20                  25                  30 

tcc cag gaa tct gtc gca gac aag atg ggg atg ggg cag tca ggc gtt      443 
Ser Gln Glu Ser Val Ala Asp Lys Met Gly Met Gly Gln Ser Gly Val 
         35                  40                  45 

ggt gct tta ttt aat ggc atc aat gca tta aat gct tat aac gcg gca      491 
Gly Ala Leu Phe Asn Gly Ile Asn Ala Leu Asn Ala Tyr Asn Ala Ala 
     50                  55                  60 

ttg cta gca aaa att ctc aaa gtt agc gtt gaa gaa ttc cat atg gat      539 
Leu Leu Ala Lys Ile Leu Lys Val Ser Val Glu Glu Phe His Met Asp 
 65                  70                  75                  80 

tat caa gtg tca agt cca atc tat gac atc aat tat tat aca tcg gag      587 
Tyr Gln Val Ser Ser Pro Ile Tyr Asp Ile Asn Tyr Tyr Thr Ser Glu 
                 85                  90                  95 

ccc tgc caa aaa atc aat gtg aag caa atc gca gcc cgc ctc ctg cct      635 
Pro Cys Gln Lys Ile Asn Val Lys Gln Ile Ala Ala Arg Leu Leu Pro 
            100                 105                 110 

ccg ctc tac tca ctg gtg ttc atc ttt ggt ttt gtg ggc aac atg ctg      683 
Pro Leu Tyr Ser Leu Val Phe Ile Phe Gly Phe Val Gly Asn Met Leu 
        115                 120                 125 

gtc atc ctc atc ctg ata aac tgc aaa agg ctg aag agc atg act gac      731 
Val Ile Leu Ile Leu Ile Asn Cys Lys Arg Leu Lys Ser Met Thr Asp 
    130                 135                 140 

atc tac ctg ctc aac ctg gcc atc tct gac ctg ttt ttc ctt ctt act      779 
Ile Tyr Leu Leu Asn Leu Ala Ile Ser Asp Leu Phe Phe Leu Leu Thr 
145                 150                 155                 160 

gtc ccc ttc tgg gct cac tat gct gcc gcc cag tgg gac ttt gga aat      827 
Val Pro Phe Trp Ala His Tyr Ala Ala Ala Gln Trp Asp Phe Gly Asn 
                165                 170                 175 

aca atg tgt caa ctc ttg aca ggg ctc tat ttt ata ggc ttc ttc tct      875 
Thr Met Cys Gln Leu Leu Thr Gly Leu Tyr Phe Ile Gly Phe Phe Ser 
            180                 185                 190 

gga atc ttc ttc atc atc ctc ctg aca atc gat agg tac ctg gct gtc      923 
Gly Ile Phe Phe Ile Ile Leu Leu Thr Ile Asp Arg Tyr Leu Ala Val 
        195                 200                 205 

gtc cat gct gtg ttt gct tta aaa gcc agg acg gtc acc ttt ggg gtg      971 
Val His Ala Val Phe Ala Leu Lys Ala Arg Thr Val Thr Phe Gly Val 
    210                 215                 220 

gtg aca agt gtg atc act tgg gtg gtg gct gtg ttt gcg tct ctc cca     1019 
Val Thr Ser Val Ile Thr Trp Val Val Ala Val Phe Ala Ser Leu Pro 
225                 230                 235                 240 

gga atc atc ttt acc aga tct caa aaa gaa ggt ctt cat tac acc tgc     1067 
Gly Ile Ile Phe Thr Arg Ser Gln Lys Glu Gly Leu His Tyr Thr Cys 
                245                 250                 255 

agc tct cat ttt cca tac agt cag tat caa ttc tgg aag aat ttc cag     1115 
Ser Ser His Phe Pro Tyr Ser Gln Tyr Gln Phe Trp Lys Asn Phe Gln 
            260                 265                 270 

aca tta aag ata gtc atc ttg ggg ctg gtc ctg ccg ctg ctt gtc atg     1163 
Thr Leu Lys Ile Val Ile Leu Gly Leu Val Leu Pro Leu Leu Val Met 
        275                 280                 285 

gtc atc tgc tac tcg gga atc cta aaa act ctg ctt cgg tgt cga aat     1211 
Val Ile Cys Tyr Ser Gly Ile Leu Lys Thr Leu Leu Arg Cys Arg Asn 
    290                 295                 300 

gag aag aag agg cac agg gct gtg agg ctt atc ttc acc atc atg att     1259 
Glu Lys Lys Arg His Arg Ala Val Arg Leu Ile Phe Thr Ile Met Ile 
305                 310                 315                 320 

gtt tat ttt ctc ttc tgg gct ccc tac aac att gtc ctt ctc ctg aac     1307 
Val Tyr Phe Leu Phe Trp Ala Pro Tyr Asn Ile Val Leu Leu Leu Asn 
                325                 330                 335 

acc ttc cag gaa ttc ttt ggc ctg aat aat tgc agt agc tct aac agg     1355 
Thr Phe Gln Glu Phe Phe Gly Leu Asn Asn Cys Ser Ser Ser Asn Arg 
            340                 345                 350 

ttg gac caa gct atg cag gtg aca gag act ctt ggg atg acg cac tgc     1403 
Leu Asp Gln Ala Met Gln Val Thr Glu Thr Leu Gly Met Thr His Cys 
        355                 360                 365 

tgc atc aac ccc atc atc tat gcc ttt gtc ggg gag aag ttc aga aac     1451 
Cys Ile Asn Pro Ile Ile Tyr Ala Phe Val Gly Glu Lys Phe Arg Asn 
    370                 375                 380 

tac ctc tta gtc ttc ttc caa aag cac att gcc aaa cgc ttc tgc aaa     1499 
Tyr Leu Leu Val Phe Phe Gln Lys His Ile Ala Lys Arg Phe Cys Lys 
385                 390                 395                 400 

tgc tgt tct att ttc cag caa gag gct ccc gag cga gca agc tca gtt     1547 
Cys Cys Ser Ile Phe Gln Gln Glu Ala Pro Glu Arg Ala Ser Ser Val 
                405                 410                 415 

tac acc cga tcc act ggg gag cag gaa ata tct gtg ggc ttg gcc tcg     1595 
Tyr Thr Arg Ser Thr Gly Glu Gln Glu Ile Ser Val Gly Leu Ala Ser 
            420                 425                 430 

agg cac cat cac cac cac cac tgaaagcttt aatgcggtag tttatcacag        1646 
Arg His His His His His His 
        435 

ttaaattgct aacgcagtca ggcaccgtgt atgaaatcta acaatgcgct catcgtcatc   1706 

ctcggcaccg tcaccctgga tgctgtaggc ataggcttgg ttatgccggt actgccgggc   1766 

ctcttgcggg atcgacgcga ggctggatgg ccttccccat tatgattctt ctcgcttccg   1826 

gcggcatcgg gatgcccgcg ttgcaggcca tgctgtccag gcaggtagat gacgaccatc   1886 

agggacagct tcaaggatcg ctcgcggctc ttaccagcct aacttcgatc actggaccgc   1946 

tgatcgtcac ggcgatttat gccgcctcgg cgagcacatg gaacgggttg gcatggattg   2006 

taggcgccgc cctatacctt gtctgcctcc ccgcgttgcg tcgcggtgca tggagccggg   2066 

ccacctcgac ctgaatggaa gccggcggca cctcgctaac ggattcacca ctccaagaat   2126 

tggagccaat caattcttgc ggagaactgt gaatgcgcaa accaaccctt ggcagaacat   2186 

atccatcgcg tccgccatct ccagcagccg cacgcggcgc atctcgggca gcgttgggtc   2246 

ctggccacgg gtgcgcatga tcgtgctcct gtcgttgagg acccggctag gctggcgggg   2306 

ttgccttact ggttagcaga atgaatcacc gatacgcgag cgaacgtgaa gcgactgctg   2366 

ctgcaaaacg tctgcgacct gagcaacaac atgaatggtc ttcggtttcc gtgtttcgta   2426 

aagtctggaa acgcggaagt cagcgccctg caccattatg ttccggatct gcatcgcagg   2486 

atgctgctgg ctaccctgtg gaacacctac atctgtatta acgaagcgct ggcattgacc   2546 

ctgagtgatt tttctctggt cccgccgcat ccataccgcc agttgtttac cctcacaacg   2606 

ttccagtaac cgggcatgtt catcatcagt aacccgtatc gtgagcatcc tctctcgttt   2666 

catcggtatc attaccccca tgaacagaaa ttccccctta cacggaggca tcaagtgacc   2726 

aaacaggaaa aaaccgccct taacatggcc cgctttatca gaagccagac attaacgctt   2786 

ctggagaaac tcaacgagct ggacgcggat gaacaggcag acatctgtga atcgcttcac   2846 

gaccacgctg atgagcttta ccgcagctgc ctcgcgcgtt tcggtgatga cggtgaaaac   2906 

ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc   2966 

agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc agccatgacc   3026 

cagtcacgta gcgatagcgg agtgtatact ggcttaacta tgcggcatca gagcagattg   3086 

tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc   3146 

gcatcaggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc   3206 

ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata   3266 

acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg   3326 

cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct   3386 

caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa   3446 

gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc   3506 

tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt   3566 

aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg   3626 

ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg   3686 

cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct   3746 

tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc   3806 

tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg   3866 

ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc   3926 

aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt   3986 

aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaccccggt   4046 

tgataatcag aaaagcccca aaaacaggaa gattgtataa gcaaatattt aaattgtaaa   4106 

cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca   4166 

ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagcccgaga tagggttgag   4226 

tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg   4286 

gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccca aatcaagttt   4346 

tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag   4406 

agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc   4466 

gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc   4526 

gcttaatgcg ccgctacagg gcgcgtaaat caatctaaag tatatatgag taaacttggt   4586 

ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt   4646 

catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat   4706 

ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag   4766 

caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct   4826 

ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt   4886 

tgcgcaacgt tgttgccatt gctgcaggca tcgtggtgtc acgctcgtcg tttggtatgg   4946 

cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca   5006 

aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt   5066 

tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat   5126 

gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac   5186 

cgagttgctc ttgcccggcg tcaacacggg ataataccgc gccacatagc agaactttaa   5246 

aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt   5306 

tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt   5366 

tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa   5426 

gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt   5486 

atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa   5546 

taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta   5606 

tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctt caagaattga   5666 

tcgatcaa                                                            5674 

 
           
             4  
             5857  
             DNA  
             Artificial Sequence  
             
               CDS  
               (300)...(1799)  
               Description of Artificial Sequence/note =
      synthetic construct  
             
           
            4 

ttctcatgtt tgacagctta tctcatcgac tgcacggtgc accaatgctt ctggcgtcag     60 

gcagccatcg gaagctgtgg tatggctgtg caggtcgtaa atcactgcat aattcgtgtc    120 

gctcaaggcg cactcccgtt ctggataatg ttttttgcgc cgacatcata acggttctgg    180 

caaatattct gaaatgagct gttgacaatt aatcatcggc tcgtataatg tggaattgtg    240 

agcggataac aattaatgtg tgaatgtgag cggatacaat ttcacacagg aaacagcgt     299 

atg agc aca aaa aag aaa cca tta aca caa gag cag ctt gag gac gca      347 
Met Ser Thr Lys Lys Lys Pro Leu Thr Gln Glu Gln Leu Glu Asp Ala 
 1               5                   10                  15 

cgt cgc ctt aaa gca att tat gaa aaa aag aaa aat gaa ctt ggc tta      395 
Arg Arg Leu Lys Ala Ile Tyr Glu Lys Lys Lys Asn Glu Leu Gly Leu 
             20                  25                  30 

tcc cag gaa tct gtc gca gac aag atg ggg atg ggg cag tca ggc gtt      443 
Ser Gln Glu Ser Val Ala Asp Lys Met Gly Met Gly Gln Ser Gly Val 
         35                  40                  45 

ggt gct tta ttt aat ggc atc aat gca tta aat gct tat aac gcg gca      491 
Gly Ala Leu Phe Asn Gly Ile Asn Ala Leu Asn Ala Tyr Asn Ala Ala 
     50                  55                  60 

ttg cta gca aaa att ctc aaa gtt agc gtt gaa gaa ttc acc atg ggg      539 
Leu Leu Ala Lys Ile Leu Lys Val Ser Val Glu Glu Phe Thr Met Gly 
 65                  70                  75                  80 

caa ccc ggg aac ggc agc gcc ttc ttg ctg gca ccc aat gga agc cat      587 
Gln Pro Gly Asn Gly Ser Ala Phe Leu Leu Ala Pro Asn Gly Ser His 
                 85                  90                  95 

gcg ccg gac cac gac gtc acg cag caa agg gac gag gtg tgg gtg gtg      635 
Ala Pro Asp His Asp Val Thr Gln Gln Arg Asp Glu Val Trp Val Val 
            100                 105                 110 

ggc atg ggc atc gtc atg tct ctc atc gtc ctg gcc atc gtg ttt ggc      683 
Gly Met Gly Ile Val Met Ser Leu Ile Val Leu Ala Ile Val Phe Gly 
        115                 120                 125 

aat gtg ctg gtc atc aca gcc att gcc aag ttc gag cgt ctg cag acg      731 
Asn Val Leu Val Ile Thr Ala Ile Ala Lys Phe Glu Arg Leu Gln Thr 
    130                 135                 140 

gtc acc aac tac ttc atc aca agc ttg gcc tgt gct gat ctg gtc atg      779 
Val Thr Asn Tyr Phe Ile Thr Ser Leu Ala Cys Ala Asp Leu Val Met 
145                 150                 155                 160 

ggg cta gca gtg gtg ccc ttt ggg gcc gcc cat att ctc atg aaa atg      827 
Gly Leu Ala Val Val Pro Phe Gly Ala Ala His Ile Leu Met Lys Met 
                165                 170                 175 

tgg act ttt ggc aac ttc tgg tgc gaa ttc tgg act tcc att gat gtg      875 
Trp Thr Phe Gly Asn Phe Trp Cys Glu Phe Trp Thr Ser Ile Asp Val 
            180                 185                 190 

ctg tgc gtc acg gca tcg att gag acc ctg tgc gtg atc gca gtc gac      923 
Leu Cys Val Thr Ala Ser Ile Glu Thr Leu Cys Val Ile Ala Val Asp 
        195                 200                 205 

cgc tac ttt gcc att act agt cct ttc aag tac cag agc ctg ctg acc      971 
Arg Tyr Phe Ala Ile Thr Ser Pro Phe Lys Tyr Gln Ser Leu Leu Thr 
    210                 215                 220 

aag aat aag gcc cgg gtg atc att ctg atg gtg tgg att gtg tca ggc     1019 
Lys Asn Lys Ala Arg Val Ile Ile Leu Met Val Trp Ile Val Ser Gly 
225                 230                 235                 240 

ctt acc tcc ttc ttg ccc att cag atg cac tgg tac agg gcc acc cac     1067 
Leu Thr Ser Phe Leu Pro Ile Gln Met His Trp Tyr Arg Ala Thr His 
                245                 250                 255 

cag gaa gcc atc aac tgc tat gcc aat gag acc tgc tgt gac ttc ttc     1115 
Gln Glu Ala Ile Asn Cys Tyr Ala Asn Glu Thr Cys Cys Asp Phe Phe 
            260                 265                 270 

acg aac caa gcc tat gcc att gcc tct tcc atc gtg tcc ttc tac gtt     1163 
Thr Asn Gln Ala Tyr Ala Ile Ala Ser Ser Ile Val Ser Phe Tyr Val 
        275                 280                 285 

ccc ctg gtg atc atg gtc ttc gtc tac tcc agg gtc ttt cag gag gcc     1211 
Pro Leu Val Ile Met Val Phe Val Tyr Ser Arg Val Phe Gln Glu Ala 
    290                 295                 300 

aaa agg cag ctc cag aag att gac aaa tct gag ggc cgc ttc cat gtc     1259 
Lys Arg Gln Leu Gln Lys Ile Asp Lys Ser Glu Gly Arg Phe His Val 
305                 310                 315                 320 

cag aac ctt agc cag gtg gag cag gat ggg cgg acg ggg cat gga ctc     1307 
Gln Asn Leu Ser Gln Val Glu Gln Asp Gly Arg Thr Gly His Gly Leu 
                325                 330                 335 

cgc aga tct tcc aag ttc tgc ttg aag gag cac aaa gcc ctc aag acg     1355 
Arg Arg Ser Ser Lys Phe Cys Leu Lys Glu His Lys Ala Leu Lys Thr 
            340                 345                 350 

tta ggc atc atc atg ggc act ttc acc ctc tgc tgg ctg ccc ttc ttc     1403 
Leu Gly Ile Ile Met Gly Thr Phe Thr Leu Cys Trp Leu Pro Phe Phe 
        355                 360                 365 

atc gtt aac att gtg cat gtg atc cag gat aac ctc atc cgt aag gaa     1451 
Ile Val Asn Ile Val His Val Ile Gln Asp Asn Leu Ile Arg Lys Glu 
    370                 375                 380 

gtt tac atc ctc cta aat tgg ata ggc tat gtc aat tct ggt ttc aat     1499 
Val Tyr Ile Leu Leu Asn Trp Ile Gly Tyr Val Asn Ser Gly Phe Asn 
385                 390                 395                 400 

ccc ctt atc tac tgc cgg agc cca gat ttc agg att gcc ttc cag gag     1547 
Pro Leu Ile Tyr Cys Arg Ser Pro Asp Phe Arg Ile Ala Phe Gln Glu 
                405                 410                 415 

ctt ctg tgc ctg cgc agg tct tct ttg aag gcc tat ggc aat ggc tac     1595 
Leu Leu Cys Leu Arg Arg Ser Ser Leu Lys Ala Tyr Gly Asn Gly Tyr 
            420                 425                 430 

tcc agc aac ggc aac aca ggg gag cag agt gga tat cac gtg gaa cag     1643 
Ser Ser Asn Gly Asn Thr Gly Glu Gln Ser Gly Tyr His Val Glu Gln 
        435                 440                 445 

gag aaa gaa aat aaa ctg ctg tgt gaa gac ctc cca ggc acg gaa gac     1691 
Glu Lys Glu Asn Lys Leu Leu Cys Glu Asp Leu Pro Gly Thr Glu Asp 
    450                 455                 460 

ttt gtg ggc cat caa ggt act gtg cct agc gat aac att gat tca caa     1739 
Phe Val Gly His Gln Gly Thr Val Pro Ser Asp Asn Ile Asp Ser Gln 
465                 470                 475                 480 

ggg agg aat tgt agt aca aat gac tca ctg cta gcc tcg agg cac cat     1787 
Gly Arg Asn Cys Ser Thr Asn Asp Ser Leu Leu Ala Ser Arg His His 
                485                 490                 495 

cac cac cac cac tgaaagcttt aatgcggtag tttatcacag ttaaattgct         1839 
His His His His 
            500 

aacgcagtca ggcaccgtgt atgaaatcta acaatgcgct catcgtcatc ctcggcaccg   1899 

tcaccctgga tgctgtaggc ataggcttgg ttatgccggt actgccgggc ctcttgcggg   1959 

atcgacgcga ggctggatgg ccttccccat tatgattctt ctcgcttccg gcggcatcgg   2019 

gatgcccgcg ttgcaggcca tgctgtccag gcaggtagat gacgaccatc agggacagct   2079 

tcaaggatcg ctcgcggctc ttaccagcct aacttcgatc actggaccgc tgatcgtcac   2139 

ggcgatttat gccgcctcgg cgagcacatg gaacgggttg gcatggattg taggcgccgc   2199 

cctatacctt gtctgcctcc ccgcgttgcg tcgcggtgca tggagccggg ccacctcgac   2259 

ctgaatggaa gccggcggca cctcgctaac ggattcacca ctccaagaat tggagccaat   2319 

caattcttgc ggagaactgt gaatgcgcaa accaaccctt ggcagaacat atccatcgcg   2379 

tccgccatct ccagcagccg cacgcggcgc atctcgggca gcgttgggtc ctggccacgg   2439 

gtgcgcatga tcgtgctcct gtcgttgagg acccggctag gctggcgggg ttgccttact   2499 

ggttagcaga atgaatcacc gatacgcgag cgaacgtgaa gcgactgctg ctgcaaaacg   2559 

tctgcgacct gagcaacaac atgaatggtc ttcggtttcc gtgtttcgta aagtctggaa   2619 

acgcggaagt cagcgccctg caccattatg ttccggatct gcatcgcagg atgctgctgg   2679 

ctaccctgtg gaacacctac atctgtatta acgaagcgct ggcattgacc ctgagtgatt   2739 

tttctctggt cccgccgcat ccataccgcc agttgtttac cctcacaacg ttccagtaac   2799 

cgggcatgtt catcatcagt aacccgtatc gtgagcatcc tctctcgttt catcggtatc   2859 

attaccccca tgaacagaaa ttccccctta cacggaggca tcaagtgacc aaacaggaaa   2919 

aaaccgccct taacatggcc cgctttatca gaagccagac attaacgctt ctggagaaac   2979 

tcaacgagct ggacgcggat gaacaggcag acatctgtga atcgcttcac gaccacgctg   3039 

atgagcttta ccgcagctgc ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca   3099 

tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc   3159 

gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc agccatgacc cagtcacgta   3219 

gcgatagcgg agtgtatact ggcttaacta tgcggcatca gagcagattg tactgagagt   3279 

gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg   3339 

ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt   3399 

atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa   3459 

gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc   3519 

gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag   3579 

gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt   3639 

gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg   3699 

aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg   3759 

ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg   3819 

taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac   3879 

tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg   3939 

gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt   3999 

taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg   4059 

tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc   4119 

tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt   4179 

ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaccccggt tgataatcag   4239 

aaaagcccca aaaacaggaa gattgtataa gcaaatattt aaattgtaaa cgttaatatt   4299 

ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa   4359 

atcggcaaaa tcccttataa atcaaaagaa tagcccgaga tagggttgag tgttgttcca   4419 

gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc   4479 

gtctatcagg gcgatggccc actacgtgaa ccatcaccca aatcaagttt tttggggtcg   4539 

aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg   4599 

ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg   4659 

gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg   4719 

ccgctacagg gcgcgtaaat caatctaaag tatatatgag taaacttggt ctgacagtta   4779 

ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt   4839 

tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag   4899 

tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca   4959 

gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc   5019 

tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt   5079 

tgttgccatt gctgcaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag   5139 

ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt   5199 

tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat   5259 

ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt   5319 

gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc   5379 

ttgcccggcg tcaacacggg ataataccgc gccacatagc agaactttaa aagtgctcat   5439 

cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag   5499 

ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt   5559 

ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg   5619 

gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta   5679 

ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc   5739 

gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt   5799 

aacctataaa aataggcgta tcacgaggcc ctttcgtctt caagaattga tcgatcaa     5857 

 
           
             5  
             5734  
             DNA  
             Artificial Sequence  
             
               CDS  
               (300)...(1676)  
               Description of Artificial Sequence/note =
      synthetic construct  
             
           
            5 

ttctcatgtt tgacagctta tctcatcgac tgcacggtgc accaatgctt ctggcgtcag     60 

gcagccatcg gaagctgtgg tatggctgtg caggtcgtaa atcactgcat aattcgtgtc    120 

gctcaaggcg cactcccgtt ctggataatg ttttttgcgc cgacatcata acggttctgg    180 

caaatattct gaaatgagct gttgacaatt aatcatcggc tcgtataatg tggaattgtg    240 

agcggataac aattaatgtg tgaatgtgag cggatacaat ttcacacagg aaacagcgt     299 

atg agc aca aaa aag aaa cca tta aca caa gag cag ctt gag gac gca      347 
Met Ser Thr Lys Lys Lys Pro Leu Thr Gln Glu Gln Leu Glu Asp Ala 
 1               5                   10                  15 

cgt cgc ctt aaa gca att tat gaa aaa aag aaa aat gaa ctt ggc tta      395 
Arg Arg Leu Lys Ala Ile Tyr Glu Lys Lys Lys Asn Glu Leu Gly Leu 
             20                  25                  30 

tcc cag gaa tct gtc gca gac aag atg ggg atg ggg cag tca ggc gtt      443 
Ser Gln Glu Ser Val Ala Asp Lys Met Gly Met Gly Gln Ser Gly Val 
         35                  40                  45 

ggt gct tta ttt aat ggc atc aat gca tta aat gct tat aac gcg gca      491 
Gly Ala Leu Phe Asn Gly Ile Asn Ala Leu Asn Ala Tyr Asn Ala Ala 
     50                  55                  60 

ttg cta gca aaa att ctc aaa gtt agc gtt gaa gaa ttc cat atg ttc      539 
Leu Leu Ala Lys Ile Leu Lys Val Ser Val Glu Glu Phe His Met Phe 
 65                  70                  75                  80 

aaa cac ctc cga aga tgg ttt atc act cac ata ttt ggg cgt tcc cgg      587 
Lys His Leu Arg Arg Trp Phe Ile Thr His Ile Phe Gly Arg Ser Arg 
                 85                  90                  95 

caa cgg gca agg ctg gtc tct aaa gaa gga aga tgt aac atc gag ttt      635 
Gln Arg Ala Arg Leu Val Ser Lys Glu Gly Arg Cys Asn Ile Glu Phe 
            100                 105                 110 

ggc aat gtg gat gca cag tca agg ttt ata ttc ttt gtg gac atc tgg      683 
Gly Asn Val Asp Ala Gln Ser Arg Phe Ile Phe Phe Val Asp Ile Trp 
        115                 120                 125 

aca act gtg ctg gac ctg aaa tgg agg tac aaa atg acc gtg ttc atc      731 
Thr Thr Val Leu Asp Leu Lys Trp Arg Tyr Lys Met Thr Val Phe Ile 
    130                 135                 140 

aca gcc ttc ttg ggg agt tgg ttc ctc ttt ggt ctc ctg tgg tat gtc      779 
Thr Ala Phe Leu Gly Ser Trp Phe Leu Phe Gly Leu Leu Trp Tyr Val 
145                 150                 155                 160 

gta gcg tat gtt cat aag gac ctc cca gag ttc tac ccg cct gac aac      827 
Val Ala Tyr Val His Lys Asp Leu Pro Glu Phe Tyr Pro Pro Asp Asn 
                165                 170                 175 

cgc act cct tgt gtg gag aac att aat ggc atg act tca gcc ttt ctg      875 
Arg Thr Pro Cys Val Glu Asn Ile Asn Gly Met Thr Ser Ala Phe Leu 
            180                 185                 190 

ttt tct cta gag act caa gtg acc ata ggt tac gga ttc agg ttt gtg      923 
Phe Ser Leu Glu Thr Gln Val Thr Ile Gly Tyr Gly Phe Arg Phe Val 
        195                 200                 205 

aca gaa cag tgc gcc act gcc att ttc ctg ctt atc ttc cag tct att      971 
Thr Glu Gln Cys Ala Thr Ala Ile Phe Leu Leu Ile Phe Gln Ser Ile 
    210                 215                 220 

ctt gga gtg atc atc aat tcc ttc atg tgt ggt gcc att tta gcc aag     1019 
Leu Gly Val Ile Ile Asn Ser Phe Met Cys Gly Ala Ile Leu Ala Lys 
225                 230                 235                 240 

atc tct aga ccc aaa aaa cgt gct aaa acc att acg ttc agc aag aat     1067 
Ile Ser Arg Pro Lys Lys Arg Ala Lys Thr Ile Thr Phe Ser Lys Asn 
                245                 250                 255 

gcg gtg atc agc aag cgt ggc ggg aag ctc tgc ctc ctc atc cga gtg     1115 
Ala Val Ile Ser Lys Arg Gly Gly Lys Leu Cys Leu Leu Ile Arg Val 
            260                 265                 270 

gcc aat ctt agg aag agc ctt ctg att ggc agc cac ata tat ggc aag     1163 
Ala Asn Leu Arg Lys Ser Leu Leu Ile Gly Ser His Ile Tyr Gly Lys 
        275                 280                 285 

ctt cta aag aca acc atc act cct gaa ggc gag acc atc att ttg gat     1211 
Leu Leu Lys Thr Thr Ile Thr Pro Glu Gly Glu Thr Ile Ile Leu Asp 
    290                 295                 300 

cag acc aac atc aac ttt gtc gtc gac gct ggc aat gaa aat ttg ttc     1259 
Gln Thr Asn Ile Asn Phe Val Val Asp Ala Gly Asn Glu Asn Leu Phe 
305                 310                 315                 320 

ttc ata tcc cca ctg acg atc tac cac att att gac cac aac agc cct     1307 
Phe Ile Ser Pro Leu Thr Ile Tyr His Ile Ile Asp His Asn Ser Pro 
                325                 330                 335 

ttc ttc cac atg gca gca gaa act ctt tcc caa cag gac ttt gag ctg     1355 
Phe Phe His Met Ala Ala Glu Thr Leu Ser Gln Gln Asp Phe Glu Leu 
            340                 345                 350 

gtg gtc ttt tta gat ggc aca gtg gaa tcc acc agt gca acc tgc cag     1403 
Val Val Phe Leu Asp Gly Thr Val Glu Ser Thr Ser Ala Thr Cys Gln 
        355                 360                 365 

gtc cgc acg tca tac gtc cca gag gag gtg ctt tgg ggc tac cgt ttc     1451 
Val Arg Thr Ser Tyr Val Pro Glu Glu Val Leu Trp Gly Tyr Arg Phe 
    370                 375                 380 

gtt cct att gtg tcc aag acc aag gaa ggg aaa tac cga gtt gat ttt     1499 
Val Pro Ile Val Ser Lys Thr Lys Glu Gly Lys Tyr Arg Val Asp Phe 
385                 390                 395                 400 

cat aac ttc ggt aag aca gtg gaa gtg gag acc cct cac tgt gcc atg     1547 
His Asn Phe Gly Lys Thr Val Glu Val Glu Thr Pro His Cys Ala Met 
                405                 410                 415 

tgc ctc tat aat gag aaa gat gcc agg gcc agg atg aag aga ggc tat     1595 
Cys Leu Tyr Asn Glu Lys Asp Ala Arg Ala Arg Met Lys Arg Gly Tyr 
            420                 425                 430 

gac aac cct aac ttt gtc ttg tca gaa gtt gat gaa acg gac gac acc     1643 
Asp Asn Pro Asn Phe Val Leu Ser Glu Val Asp Glu Thr Asp Asp Thr 
        435                 440                 445 

cag atg gcc tcg agg cac cat cac cac cac cac tgaaagcttt aatgcggtag   1696 
Gln Met Ala Ser Arg His His His His His His 
    450                 455 

tttatcacag ttaaattgct aacgcagtca ggcaccgtgt atgaaatcta acaatgcgct   1756 

catcgtcatc ctcggcaccg tcaccctgga tgctgtaggc ataggcttgg ttatgccggt   1816 

actgccgggc ctcttgcggg atcgacgcga ggctggatgg ccttccccat tatgattctt   1876 

ctcgcttccg gcggcatcgg gatgcccgcg ttgcaggcca tgctgtccag gcaggtagat   1936 

gacgaccatc agggacagct tcaaggatcg ctcgcggctc ttaccagcct aacttcgatc   1996 

actggaccgc tgatcgtcac ggcgatttat gccgcctcgg cgagcacatg gaacgggttg   2056 

gcatggattg taggcgccgc cctatacctt gtctgcctcc ccgcgttgcg tcgcggtgca   2116 

tggagccggg ccacctcgac ctgaatggaa gccggcggca cctcgctaac ggattcacca   2176 

ctccaagaat tggagccaat caattcttgc ggagaactgt gaatgcgcaa accaaccctt   2236 

ggcagaacat atccatcgcg tccgccatct ccagcagccg cacgcggcgc atctcgggca   2296 

gcgttgggtc ctggccacgg gtgcgcatga tcgtgctcct gtcgttgagg acccggctag   2356 

gctggcgggg ttgccttact ggttagcaga atgaatcacc gatacgcgag cgaacgtgaa   2416 

gcgactgctg ctgcaaaacg tctgcgacct gagcaacaac atgaatggtc ttcggtttcc   2476 

gtgtttcgta aagtctggaa acgcggaagt cagcgccctg caccattatg ttccggatct   2536 

gcatcgcagg atgctgctgg ctaccctgtg gaacacctac atctgtatta acgaagcgct   2596 

ggcattgacc ctgagtgatt tttctctggt cccgccgcat ccataccgcc agttgtttac   2656 

cctcacaacg ttccagtaac cgggcatgtt catcatcagt aacccgtatc gtgagcatcc   2716 

tctctcgttt catcggtatc attaccccca tgaacagaaa ttccccctta cacggaggca   2776 

tcaagtgacc aaacaggaaa aaaccgccct taacatggcc cgctttatca gaagccagac   2836 

attaacgctt ctggagaaac tcaacgagct ggacgcggat gaacaggcag acatctgtga   2896 

atcgcttcac gaccacgctg atgagcttta ccgcagctgc ctcgcgcgtt tcggtgatga   2956 

cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga   3016 

tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc   3076 

agccatgacc cagtcacgta gcgatagcgg agtgtatact ggcttaacta tgcggcatca   3136 

gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg   3196 

agaaaatacc gcatcaggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc   3256 

gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa   3316 

tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt   3376 

aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa   3436 

aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt   3496 

ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg   3556 

tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc   3616 

agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc   3676 

gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta   3736 

tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct   3796 

acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc   3856 

tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa   3916 

caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa   3976 

aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa   4036 

aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt   4096 

ttaccccggt tgataatcag aaaagcccca aaaacaggaa gattgtataa gcaaatattt   4156 

aaattgtaaa cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat   4216 

tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagcccgaga   4276 

tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca   4336 

acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccca   4396 

aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc   4456 

cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag   4516 

cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca   4576 

cacccgccgc gcttaatgcg ccgctacagg gcgcgtaaat caatctaaag tatatatgag   4636 

taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt   4696 

ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag   4756 

ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca   4816 

gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact   4876 

ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca   4936 

gttaatagtt tgcgcaacgt tgttgccatt gctgcaggca tcgtggtgtc acgctcgtcg   4996 

tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc   5056 

atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg   5116 

gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca   5176 

tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt   5236 

atgcggcgac cgagttgctc ttgcccggcg tcaacacggg ataataccgc gccacatagc   5296 

agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc   5356 

ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca   5416 

tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa   5476 

aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat   5536 

tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa   5596 

aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa   5656 

accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctt   5716 

caagaattga tcgatcaa                                                 5734 

 
           
             6  
             5197  
             DNA  
             Artificial Sequence  
             
               CDS  
               (300)...(1139)  
               Description of Artificial Sequence/note =
      synthetic construct  
             
           
            6 

ttctcatgtt tgacagctta tctcatcgac tgcacggtgc accaatgctt ctggcgtcag     60 

gcagccatcg gaagctgtgg tatggctgtg caggtcgtaa atcactgcat aattcgtgtc    120 

gctcaaggcg cactcccgtt ctggataatg ttttttgcgc cgacatcata acggttctgg    180 

caaatattct gaaatgagct gttgacaatt aatcatcggc tcgtataatg tggaattgtg    240 

agcggataac aattaatgtg tgaatgtgag cggatacaat ttcacacagg aaacagcgt     299 

atg agc aca aaa aag aaa cca tta aca caa gag cag ctt gag gac gca      347 
Met Ser Thr Lys Lys Lys Pro Leu Thr Gln Glu Gln Leu Glu Asp Ala 
 1               5                   10                  15 

cgt cgc ctt aaa gca att tat gaa aaa aag aaa aat gaa ctt ggc tta      395 
Arg Arg Leu Lys Ala Ile Tyr Glu Lys Lys Lys Asn Glu Leu Gly Leu 
             20                  25                  30 

tcc cag gaa tct gtc gca gac aag atg ggg atg ggg cag tca ggc gtt      443 
Ser Gln Glu Ser Val Ala Asp Lys Met Gly Met Gly Gln Ser Gly Val 
         35                  40                  45 

ggt gct tta ttt aat ggc atc aat gca tta aat gct tat aac gcg gca      491 
Gly Ala Leu Phe Asn Gly Ile Asn Ala Leu Asn Ala Tyr Asn Ala Ala 
     50                  55                  60 

ttg cta gca aaa att ctc aaa gtt agc gtt gaa gaa ttc cat atg gct      539 
Leu Leu Ala Lys Ile Leu Lys Val Ser Val Glu Glu Phe His Met Ala 
 65                  70                  75                  80 

gcc atc cgg aag aaa ctg gtg att gtt ggt gat gga gcc tgt gga aag      587 
Ala Ile Arg Lys Lys Leu Val Ile Val Gly Asp Gly Ala Cys Gly Lys 
                 85                  90                  95 

aca tgc ttg ctc ata gtc ttc agc aag gac cag ttc cca gag gtg tat      635 
Thr Cys Leu Leu Ile Val Phe Ser Lys Asp Gln Phe Pro Glu Val Tyr 
            100                 105                 110 

gtg ccc aca gtg ttt gag aac tat gtg gca gat atc gag gtg gat gga      683 
Val Pro Thr Val Phe Glu Asn Tyr Val Ala Asp Ile Glu Val Asp Gly 
        115                 120                 125 

aag cag gta gag ttg gct ttg tgg gac aca gct ggg cag gaa gat tat      731 
Lys Gln Val Glu Leu Ala Leu Trp Asp Thr Ala Gly Gln Glu Asp Tyr 
    130                 135                 140 

gat cgc ctg agg ccc ctc tcc tac cca gat acc gat gtt ata ctg atg      779 
Asp Arg Leu Arg Pro Leu Ser Tyr Pro Asp Thr Asp Val Ile Leu Met 
145                 150                 155                 160 

tgt ttt tcc atc gac agc cct gat agt tta gaa aac atc cca gaa aag      827 
Cys Phe Ser Ile Asp Ser Pro Asp Ser Leu Glu Asn Ile Pro Glu Lys 
                165                 170                 175 

tgg acc cca gaa gtc aag cat ttc tgt ccc aac gtg ccc atc atc ctg      875 
Trp Thr Pro Glu Val Lys His Phe Cys Pro Asn Val Pro Ile Ile Leu 
            180                 185                 190 

gtt ggg aat aag aag gat ctt cgg aat gat gag cac aca agg cgg gag      923 
Val Gly Asn Lys Lys Asp Leu Arg Asn Asp Glu His Thr Arg Arg Glu 
        195                 200                 205 

cta gcc aag atg aag cag gag ccg gtg aaa cct gaa gaa ggc aga gat      971 
Leu Ala Lys Met Lys Gln Glu Pro Val Lys Pro Glu Glu Gly Arg Asp 
    210                 215                 220 

atg gca aac agg att ggc gct ttt ggg tac atg gag tgt tca gca aag     1019 
Met Ala Asn Arg Ile Gly Ala Phe Gly Tyr Met Glu Cys Ser Ala Lys 
225                 230                 235                 240 

acc aaa gat gga gtg aga gag gtt ttt gaa atg gct acg aga gct gct     1067 
Thr Lys Asp Gly Val Arg Glu Val Phe Glu Met Ala Thr Arg Ala Ala 
                245                 250                 255 

ctg caa gct aga cgt ggg aag aaa aaa tct ggt tgc ctt gtc ttg gcc     1115 
Leu Gln Ala Arg Arg Gly Lys Lys Lys Ser Gly Cys Leu Val Leu Ala 
            260                 265                 270 

tcg agg cac cat cac cac cac cac tgaaagcttt aatgcggtag tttatcacag    1169 
Ser Arg His His His His His His 
        275                 280 

ttaaattgct aacgcagtca ggcaccgtgt atgaaatcta acaatgcgct catcgtcatc   1229 

ctcggcaccg tcaccctgga tgctgtaggc ataggcttgg ttatgccggt actgccgggc   1289 

ctcttgcggg atcgacgcga ggctggatgg ccttccccat tatgattctt ctcgcttccg   1349 

gcggcatcgg gatgcccgcg ttgcaggcca tgctgtccag gcaggtagat gacgaccatc   1409 

agggacagct tcaaggatcg ctcgcggctc ttaccagcct aacttcgatc actggaccgc   1469 

tgatcgtcac ggcgatttat gccgcctcgg cgagcacatg gaacgggttg gcatggattg   1529 

taggcgccgc cctatacctt gtctgcctcc ccgcgttgcg tcgcggtgca tggagccggg   1589 

ccacctcgac ctgaatggaa gccggcggca cctcgctaac ggattcacca ctccaagaat   1649 

tggagccaat caattcttgc ggagaactgt gaatgcgcaa accaaccctt ggcagaacat   1709 

atccatcgcg tccgccatct ccagcagccg cacgcggcgc atctcgggca gcgttgggtc   1769 

ctggccacgg gtgcgcatga tcgtgctcct gtcgttgagg acccggctag gctggcgggg   1829 

ttgccttact ggttagcaga atgaatcacc gatacgcgag cgaacgtgaa gcgactgctg   1889 

ctgcaaaacg tctgcgacct gagcaacaac atgaatggtc ttcggtttcc gtgtttcgta   1949 

aagtctggaa acgcggaagt cagcgccctg caccattatg ttccggatct gcatcgcagg   2009 

atgctgctgg ctaccctgtg gaacacctac atctgtatta acgaagcgct ggcattgacc   2069 

ctgagtgatt tttctctggt cccgccgcat ccataccgcc agttgtttac cctcacaacg   2129 

ttccagtaac cgggcatgtt catcatcagt aacccgtatc gtgagcatcc tctctcgttt   2189 

catcggtatc attaccccca tgaacagaaa ttccccctta cacggaggca tcaagtgacc   2249 

aaacaggaaa aaaccgccct taacatggcc cgctttatca gaagccagac attaacgctt   2309 

ctggagaaac tcaacgagct ggacgcggat gaacaggcag acatctgtga atcgcttcac   2369 

gaccacgctg atgagcttta ccgcagctgc ctcgcgcgtt tcggtgatga cggtgaaaac   2429 

ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc   2489 

agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc agccatgacc   2549 

cagtcacgta gcgatagcgg agtgtatact ggcttaacta tgcggcatca gagcagattg   2609 

tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc   2669 

gcatcaggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc   2729 

ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata   2789 

acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg   2849 

cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct   2909 

caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa   2969 

gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc   3029 

tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt   3089 

aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg   3149 

ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg   3209 

cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct   3269 

tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc   3329 

tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg   3389 

ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc   3449 

aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt   3509 

aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaccccggt   3569 

tgataatcag aaaagcccca aaaacaggaa gattgtataa gcaaatattt aaattgtaaa   3629 

cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca   3689 

ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagcccgaga tagggttgag   3749 

tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg   3809 

gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccca aatcaagttt   3869 

tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag   3929 

agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc   3989 

gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc   4049 

gcttaatgcg ccgctacagg gcgcgtaaat caatctaaag tatatatgag taaacttggt   4109 

ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt   4169 

catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat   4229 

ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag   4289 

caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct   4349 

ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt   4409 

tgcgcaacgt tgttgccatt gctgcaggca tcgtggtgtc acgctcgtcg tttggtatgg   4469 

cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca   4529 

aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt   4589 

tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat   4649 

gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac   4709 

cgagttgctc ttgcccggcg tcaacacggg ataataccgc gccacatagc agaactttaa   4769 

aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt   4829 

tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt   4889 

tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa   4949 

gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt   5009 

atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa   5069 

taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta   5129 

tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctt caagaattga   5189 

tcgatcaa                                                            5197 

 
           
             7  
             5688  
             DNA  
             Artificial Sequence  
             
               CDS  
               (300)...(1631)  
               Description of Artificial Sequence/note =
      synthetic construct  
             
           
            7 

ttctcatgtt tgacagctta tctcatcgac tgcacggtgc accaatgctt ctggcgtcag     60 

gcagccatcg gaagctgtgg tatggctgtg caggtcgtaa atcactgcat aattcgtgtc    120 

gctcaaggcg cactcccgtt ctggataatg ttttttgcgc cgacatcata acggttctgg    180 

caaatattct gaaatgagct gttgacaatt aatcatcggc tcgtataatg tggaattgtg    240 

agcggataac aattaatgtg tgaatgtgag cggatacaat ttcacacagg aaacagcgt     299 

atg agc aca aaa aag aaa cca tta aca caa gag cag ctt gag gac gca      347 
Met Ser Thr Lys Lys Lys Pro Leu Thr Gln Glu Gln Leu Glu Asp Ala 
 1               5                   10                  15 

cgt cgc ctt aaa gca att tat gaa aaa aag aaa aat gaa ctt ggc tta      395 
Arg Arg Leu Lys Ala Ile Tyr Glu Lys Lys Lys Asn Glu Leu Gly Leu 
             20                  25                  30 

tcc cag gaa tct gtc gca gac aag atg ggg atg ggg cag tca ggc gtt      443 
Ser Gln Glu Ser Val Ala Asp Lys Met Gly Met Gly Gln Ser Gly Val 
         35                  40                  45 

ggt gct tta ttt aat ggc atc aat gca tta aat gct tat aac gcg gca      491 
Gly Ala Leu Phe Asn Gly Ile Asn Ala Leu Asn Ala Tyr Asn Ala Ala 
     50                  55                  60 

ttg cta gca aaa att ctc aaa gtt agc gtt gaa gaa ttc gca gct cat      539 
Leu Leu Ala Lys Ile Leu Lys Val Ser Val Glu Glu Phe Ala Ala His 
 65                  70                  75                  80 

atg aag gag acg cgg ggc gac gga ggg agc gcc ccc ttc tgc acc cgc      587 
Met Lys Glu Thr Arg Gly Asp Gly Gly Ser Ala Pro Phe Cys Thr Arg 
                 85                  90                  95 

ctc aac cac tcg tat cca ggc atg tgg gcg ccc gag gca cgg ggc aac      635 
Leu Asn His Ser Tyr Pro Gly Met Trp Ala Pro Glu Ala Arg Gly Asn 
            100                 105                 110 

ctc aca cgc ccc cca ggg ccc ggc gag gac tgt ggc tcg gtg tcc gtg      683 
Leu Thr Arg Pro Pro Gly Pro Gly Glu Asp Cys Gly Ser Val Ser Val 
        115                 120                 125 

gcc ttc ccg atc acc atg ctg atc acc ggc ttc gtg ggc aac gcg ctg      731 
Ala Phe Pro Ile Thr Met Leu Ile Thr Gly Phe Val Gly Asn Ala Leu 
    130                 135                 140 

gcc atg ctg ctc gtg tcg cgt agc tac cgg cgt cgg gag agc aag cgc      779 
Ala Met Leu Leu Val Ser Arg Ser Tyr Arg Arg Arg Glu Ser Lys Arg 
145                 150                 155                 160 

aag aag tcg ttc ctg ttg tgc atc ggc tgg ctg gcg ctc act gac ctg      827 
Lys Lys Ser Phe Leu Leu Cys Ile Gly Trp Leu Ala Leu Thr Asp Leu 
                165                 170                 175 

gtc ggg cag ctg ctc aca agc ccc gtg gtc atc ttg gtg tac cta tcc      875 
Val Gly Gln Leu Leu Thr Ser Pro Val Val Ile Leu Val Tyr Leu Ser 
            180                 185                 190 

aag cag cgc tgg gag cag ctc gac ccg tcg ggg cgc ctg tgc acc ttc      923 
Lys Gln Arg Trp Glu Gln Leu Asp Pro Ser Gly Arg Leu Cys Thr Phe 
        195                 200                 205 

ttt ggt ctg acc atg act gtt ttc ggg ctg tcc tcg ctc ttc atc gcc      971 
Phe Gly Leu Thr Met Thr Val Phe Gly Leu Ser Ser Leu Phe Ile Ala 
    210                 215                 220 

agc gcc atg gct gtc gag agg gcg ctg gcc atc cgt gcg cca cac tgg     1019 
Ser Ala Met Ala Val Glu Arg Ala Leu Ala Ile Arg Ala Pro His Trp 
225                 230                 235                 240 

tac gcg agc cac atg aag acg cgt gcc act cgc gcc gtc ctg ctg ggc     1067 
Tyr Ala Ser His Met Lys Thr Arg Ala Thr Arg Ala Val Leu Leu Gly 
                245                 250                 255 

gtg tgg ctg gca gtg ctc gcc ttc gcc ctg cta cct gtg ctg ggt gtg     1115 
Val Trp Leu Ala Val Leu Ala Phe Ala Leu Leu Pro Val Leu Gly Val 
            260                 265                 270 

ggt cag tac acc atc cag tgg ccc ggg acg tgg tgc ttc atc agc acc     1163 
Gly Gln Tyr Thr Ile Gln Trp Pro Gly Thr Trp Cys Phe Ile Ser Thr 
        275                 280                 285 

gga cga ggg gac aac ggg acg agc tct tca cac aac tgg ggc aac ctt     1211 
Gly Arg Gly Asp Asn Gly Thr Ser Ser Ser His Asn Trp Gly Asn Leu 
    290                 295                 300 

ttc ttc gcc tcc acc ttt gcc ttc ctg ggc ctc ttg gcg ctg gcc atc     1259 
Phe Phe Ala Ser Thr Phe Ala Phe Leu Gly Leu Leu Ala Leu Ala Ile 
305                 310                 315                 320 

acc ttc acc tgc aac ctg gcc acc att aag gct ctg gtg tcc cgc tgc     1307 
Thr Phe Thr Cys Asn Leu Ala Thr Ile Lys Ala Leu Val Ser Arg Cys 
                325                 330                 335 

cgg gca aag gcg gca gca tca cag tcc agt gcc cag tgg ggc cgg atc     1355 
Arg Ala Lys Ala Ala Ala Ser Gln Ser Ser Ala Gln Trp Gly Arg Ile 
            340                 345                 350 

acg acc gag acg gcc atc cag ctc atg ggg atc atg tgc gtg ctg tcg     1403 
Thr Thr Glu Thr Ala Ile Gln Leu Met Gly Ile Met Cys Val Leu Ser 
        355                 360                 365 

gtc tgc tgg tcg ccc cta ctg ata atg atg ttg aaa atg atc ttc aat     1451 
Val Cys Trp Ser Pro Leu Leu Ile Met Met Leu Lys Met Ile Phe Asn 
    370                 375                 380 

cag aca tca gtt gag cac tgc aag aca gac aca gga aag cag aaa gaa     1499 
Gln Thr Ser Val Glu His Cys Lys Thr Asp Thr Gly Lys Gln Lys Glu 
385                 390                 395                 400 

tgc aac ttc ttc tta ata gct gtt cgc ctg gct tca ctg aac cag ata     1547 
Cys Asn Phe Phe Leu Ile Ala Val Arg Leu Ala Ser Leu Asn Gln Ile 
                405                 410                 415 

ttg gat ccc tgg gtt tat ctg ctg cta aga aag att ctt ctt cgg aag     1595 
Leu Asp Pro Trp Val Tyr Leu Leu Leu Arg Lys Ile Leu Leu Arg Lys 
            420                 425                 430 

ttt tgc cag gcc tcg agg cac cat cac cac cac cac tgaagcttta          1641 
Phe Cys Gln Ala Ser Arg His His His His His His 
        435                 440 

atgcggtagt ttatcacagt taaattgcta acgcagtcag gcaccgtgta tgaaatctaa   1701 

caatgcgctc atcgtcatcc tcggcaccgt caccctggat gctgtaggca taggcttggt   1761 

tatgccggta ctgccgggcc tcttgcggga tcgacgcgag gctggatggc cttccccatt   1821 

atgattcttc tcgcttccgg cggcatcggg atgcccgcgt tgcaggccat gctgtccagg   1881 

caggtagatg acgaccatca gggacagctt caaggatcgc tcgcggctct taccagccta   1941 

acttcgatca ctggaccgct gatcgtcacg gcgatttatg ccgcctcggc gagcacatgg   2001 

aacgggttgg catggattgt aggcgccgcc ctataccttg tctgcctccc cgcgttgcgt   2061 

cgcggtgcat ggagccgggc cacctcgacc tgaatggaag ccggcggcac ctcgctaacg   2121 

gattcaccac tccaagaatt ggagccaatc aattcttgcg gagaactgtg aatgcgcaaa   2181 

ccaacccttg gcagaacata tccatcgcgt ccgccatctc cagcagccgc acgcggcgca   2241 

tctcgggcag cgttgggtcc tggccacggg tgcgcatgat cgtgctcctg tcgttgagga   2301 

cccggctagg ctggcggggt tgccttactg gttagcagaa tgaatcaccg atacgcgagc   2361 

gaacgtgaag cgactgctgc tgcaaaacgt ctgcgacctg agcaacaaca tgaatggtct   2421 

tcggtttccg tgtttcgtaa agtctggaaa cgcggaagtc agcgccctgc accattatgt   2481 

tccggatctg catcgcagga tgctgctggc taccctgtgg aacacctaca tctgtattaa   2541 

cgaagcgctg gcattgaccc tgagtgattt ttctctggtc ccgccgcatc cataccgcca   2601 

gttgtttacc ctcacaacgt tccagtaacc gggcatgttc atcatcagta acccgtatcg   2661 

tgagcatcct ctctcgtttc atcggtatca ttacccccat gaacagaaat tcccccttac   2721 

acggaggcat caagtgacca aacaggaaaa aaccgccctt aacatggccc gctttatcag   2781 

aagccagaca ttaacgcttc tggagaaact caacgagctg gacgcggatg aacaggcaga   2841 

catctgtgaa tcgcttcacg accacgctga tgagctttac cgcagctgcc tcgcgcgttt   2901 

cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct   2961 

gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg   3021 

tcggggcgca gccatgaccc agtcacgtag cgatagcgga gtgtatactg gcttaactat   3081 

gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga   3141 

tgcgtaagga gaaaataccg catcaggcgc tcttccgctt cctcgctcac tgactcgctg   3201 

cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta   3261 

tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc   3321 

aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag   3381 

catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac   3441 

caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc   3501 

ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt   3561 

aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc   3621 

gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga   3681 

cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta   3741 

ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta   3801 

tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga   3861 

tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg   3921 

cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag   3981 

tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc   4041 

tagatccttt taccccggtt gataatcaga aaagccccaa aaacaggaag attgtataag   4101 

caaatattta aattgtaaac gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa   4161 

tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat   4221 

agcccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg   4281 

tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac   4341 

catcacccaa atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta   4401 

aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag   4461 

ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg   4521 

taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtaaatc aatctaaagt   4581 

atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca   4641 

gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg   4701 

atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca   4761 

ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt   4821 

cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt   4881 

agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctgcaggcat cgtggtgtca   4941 

cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca   5001 

tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga   5061 

agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact   5121 

gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga   5181 

gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caacacggga taataccgcg   5241 

ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc   5301 

tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga   5361 

tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat   5421 

gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt   5481 

caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt   5541 

atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac   5601 

gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc   5661 

tttcgtcttc aagaattgat cgatcaa                                       5688 

 
           
             8  
             6587  
             DNA  
             Artificial Sequence  
             
               CDS  
               (300)...(2267)  
               Description of Artificial Sequence/note =
      synthetic construct  
             
           
            8 

ttctcatgtt tgacagctta tctcatcgac tgcacggtgc accaatgctt ctggcgtcag     60 

gcagccatcg gaagctgtgg tatggctgtg caggtcgtaa atcactgcat aattcgtgtc    120 

gctcaaggcg cactcccgtt ctggataatg ttttttgcgc cgacatcata acggttctgg    180 

caaatattct gaaatgagct gttgacaatt aatcatcggc tcgtataatg tggaattgtg    240 

agcggataac aattaatgtg tgaatgtgag cggatacaat ttcacacagg aaacagcgt     299 

atg agc aca aaa aag aaa cca tta aca caa gag cag ctt gag gac gca      347 
Met Ser Thr Lys Lys Lys Pro Leu Thr Gln Glu Gln Leu Glu Asp Ala 
 1               5                   10                  15 

cgt cgc ctt aaa gca att tat gaa aaa aag aaa aat gaa ctt ggc tta      395 
Arg Arg Leu Lys Ala Ile Tyr Glu Lys Lys Lys Asn Glu Leu Gly Leu 
             20                  25                  30 

tcc cag gaa tct gtc gca gac aag atg ggg atg ggg cag tca ggc gtt      443 
Ser Gln Glu Ser Val Ala Asp Lys Met Gly Met Gly Gln Ser Gly Val 
         35                  40                  45 

ggt gct tta ttt aat ggc atc aat gca tta aat gct tat aac gcg gca      491 
Gly Ala Leu Phe Asn Gly Ile Asn Ala Leu Asn Ala Tyr Asn Ala Ala 
     50                  55                  60 

ttg cta gca aaa att ctc aaa gtt agc gtt gaa gaa ttc gca gct cat      539 
Leu Leu Ala Lys Ile Leu Lys Val Ser Val Glu Glu Phe Ala Ala His 
 65                  70                  75                  80 

atg aag gag acg cgg ggc gac gga ggg agc gcc ccc ttc tgc acc cgc      587 
Met Lys Glu Thr Arg Gly Asp Gly Gly Ser Ala Pro Phe Cys Thr Arg 
                 85                  90                  95 

ctc aac cac tcg tat cca ggc atg tgg gcg ccc gag gca cgg ggc aac      635 
Leu Asn His Ser Tyr Pro Gly Met Trp Ala Pro Glu Ala Arg Gly Asn 
            100                 105                 110 

ctc aca cgc ccc cca ggg ccc ggc gag gac tgt ggc tcg gtg tcc gtg      683 
Leu Thr Arg Pro Pro Gly Pro Gly Glu Asp Cys Gly Ser Val Ser Val 
        115                 120                 125 

gcc ttc ccg atc acc atg ctg atc acc ggc ttc gtg ggc aac gcg ctg      731 
Ala Phe Pro Ile Thr Met Leu Ile Thr Gly Phe Val Gly Asn Ala Leu 
    130                 135                 140 

gcc atg ctg ctc gtg tcg cgt agc tac cgg cgt cgg gag agc aag cgc      779 
Ala Met Leu Leu Val Ser Arg Ser Tyr Arg Arg Arg Glu Ser Lys Arg 
145                 150                 155                 160 

aag aag tcg ttc ctg ttg tgc atc ggc tgg ctg gcg ctc act gac ctg      827 
Lys Lys Ser Phe Leu Leu Cys Ile Gly Trp Leu Ala Leu Thr Asp Leu 
                165                 170                 175 

gtc ggg cag ctg ctc aca agc ccc gtg gtc atc ttg gtg tac cta tcc      875 
Val Gly Gln Leu Leu Thr Ser Pro Val Val Ile Leu Val Tyr Leu Ser 
            180                 185                 190 

aag cag cgc tgg gag cag ctc gac ccg tcg ggg cgc ctg tgc acc ttc      923 
Lys Gln Arg Trp Glu Gln Leu Asp Pro Ser Gly Arg Leu Cys Thr Phe 
        195                 200                 205 

ttt ggt ctg acc atg act gtt ttc ggg ctg tcc tcg ctc ttc atc gcc      971 
Phe Gly Leu Thr Met Thr Val Phe Gly Leu Ser Ser Leu Phe Ile Ala 
    210                 215                 220 

agc gcc atg gct gtc gag agg gcg ctg gcc atc cgt gcg cca cac tgg     1019 
Ser Ala Met Ala Val Glu Arg Ala Leu Ala Ile Arg Ala Pro His Trp 
225                 230                 235                 240 

tac gcg agc cac atg aag acg cgt gcc act cgc gcc gtc ctg ctg ggc     1067 
Tyr Ala Ser His Met Lys Thr Arg Ala Thr Arg Ala Val Leu Leu Gly 
                245                 250                 255 

gtg tgg ctg gca gtg ctc gcc ttc gcc ctg cta cct gtg ctg ggt gtg     1115 
Val Trp Leu Ala Val Leu Ala Phe Ala Leu Leu Pro Val Leu Gly Val 
            260                 265                 270 

ggt cag tac acc atc cag tgg ccc ggg acg tgg tgc ttc atc agc acc     1163 
Gly Gln Tyr Thr Ile Gln Trp Pro Gly Thr Trp Cys Phe Ile Ser Thr 
        275                 280                 285 

gga cga ggg gac aac ggg acg agc tct tca cac aac tgg ggc aac ctt     1211 
Gly Arg Gly Asp Asn Gly Thr Ser Ser Ser His Asn Trp Gly Asn Leu 
    290                 295                 300 

ttc ttc gcc tcc acc ttt gcc ttc ctg ggc ctc ttg gcg ctg gcc atc     1259 
Phe Phe Ala Ser Thr Phe Ala Phe Leu Gly Leu Leu Ala Leu Ala Ile 
305                 310                 315                 320 

acc ttc acc tgc aac ctg gcc acc att aag gct ctg gtg tcc cgc tgc     1307 
Thr Phe Thr Cys Asn Leu Ala Thr Ile Lys Ala Leu Val Ser Arg Cys 
                325                 330                 335 

cgg gca aag gcg gca gca tca cag tcc agt gcc cag tgg ggc cgg atc     1355 
Arg Ala Lys Ala Ala Ala Ser Gln Ser Ser Ala Gln Trp Gly Arg Ile 
            340                 345                 350 

acg acc gag acg gcc atc cag ctc atg ggg atc atg tgc gtg ctg tcg     1403 
Thr Thr Glu Thr Ala Ile Gln Leu Met Gly Ile Met Cys Val Leu Ser 
        355                 360                 365 

gtc tgc tgg tcg ccc cta ctg ata atg atg ttg aaa atg atc ttc aat     1451 
Val Cys Trp Ser Pro Leu Leu Ile Met Met Leu Lys Met Ile Phe Asn 
    370                 375                 380 

cag aca tca gtt gag cac tgc aag aca gac aca gga aag cag aaa gaa     1499 
Gln Thr Ser Val Glu His Cys Lys Thr Asp Thr Gly Lys Gln Lys Glu 
385                 390                 395                 400 

tgc aac ttc ttc tta ata gct gtt cgc ctg gct tca ctg aac cag ata     1547 
Cys Asn Phe Phe Leu Ile Ala Val Arg Leu Ala Ser Leu Asn Gln Ile 
                405                 410                 415 

ttg gat ccc tgg gtt tat ctg ctg cta aga aag att ctt ctt cgg aag     1595 
Leu Asp Pro Trp Val Tyr Leu Leu Leu Arg Lys Ile Leu Leu Arg Lys 
            420                 425                 430 

ttt tgc cag gta att cat gaa aat aat gag cag aag gat gaa att cag     1643 
Phe Cys Gln Val Ile His Glu Asn Asn Glu Gln Lys Asp Glu Ile Gln 
        435                 440                 445 

cgt gag aac agg aac gtc tca cac agt ggg caa cac gaa gag gcc aga     1691 
Arg Glu Asn Arg Asn Val Ser His Ser Gly Gln His Glu Glu Ala Arg 
    450                 455                 460 

gac agt gag aag agc aaa acc atc cct ggc ctg ttc tcc att ctg ctg     1739 
Asp Ser Glu Lys Ser Lys Thr Ile Pro Gly Leu Phe Ser Ile Leu Leu 
465                 470                 475                 480 

cag gct gac cct ggt gct cgt cct tat cag caa gcc tcg agc ctg gtg     1787 
Gln Ala Asp Pro Gly Ala Arg Pro Tyr Gln Gln Ala Ser Ser Leu Val 
                485                 490                 495 

cca cgc gga tcc gtt cga gaa atc tac gag atg tat gaa gcg gtt agc     1835 
Pro Arg Gly Ser Val Arg Glu Ile Tyr Glu Met Tyr Glu Ala Val Ser 
            500                 505                 510 

atg cag ccg tca ctt aga agt gag tat gag tac cct gtt ttt tct cat     1883 
Met Gln Pro Ser Leu Arg Ser Glu Tyr Glu Tyr Pro Val Phe Ser His 
        515                 520                 525 

gtt cag gca ggg atg ttc tca cct aag ctt aga acc ttt acc aaa ggt     1931 
Val Gln Ala Gly Met Phe Ser Pro Lys Leu Arg Thr Phe Thr Lys Gly 
    530                 535                 540 

gat gcg gag aga tgg gta agc aca acc aaa aaa gcc agt gat tct gca     1979 
Asp Ala Glu Arg Trp Val Ser Thr Thr Lys Lys Ala Ser Asp Ser Ala 
545                 550                 555                 560 

ttc tgg ctt gag gtt gaa ggt aat tcc atg acc gca cca aca ggc tcc     2027 
Phe Trp Leu Glu Val Glu Gly Asn Ser Met Thr Ala Pro Thr Gly Ser 
                565                 570                 575 

aag cca agc ttt cct gac gga atg tta att ctc gtt gac cct gag cag     2075 
Lys Pro Ser Phe Pro Asp Gly Met Leu Ile Leu Val Asp Pro Glu Gln 
            580                 585                 590 

gct gtt gag cca ggt gat ttc tgc ata gcc aga ctt ggg ggt gat gag     2123 
Ala Val Glu Pro Gly Asp Phe Cys Ile Ala Arg Leu Gly Gly Asp Glu 
        595                 600                 605 

ttt acc ttc aag aaa ctg atc agg gat agc ggt cag gtg ttt tta caa     2171 
Phe Thr Phe Lys Lys Leu Ile Arg Asp Ser Gly Gln Val Phe Leu Gln 
    610                 615                 620 

cca cta aac cca cag tac cca atg atc cca tgc aat gag agt tgt tcc     2219 
Pro Leu Asn Pro Gln Tyr Pro Met Ile Pro Cys Asn Glu Ser Cys Ser 
625                 630                 635                 640 

gtt gtg ggg aaa gtt atc gct agt cag tgg cct gaa gag acg ttt ggc     2267 
Val Val Gly Lys Val Ile Ala Ser Gln Trp Pro Glu Glu Thr Phe Gly 
                645                 650                 655 

tgatcggcaa ggtgttctgg tcggcgcata gctgataaca attgagcaag aatcttcatc   2327 

gaattagggg aattttcact cccctcagaa cataacatag taaatggatt gaattatgaa   2387 

gaatggtttt tatgcgactt accgcagcaa aaataaaggg aaagataagc gctcaataaa   2447 

cctgtctgtt ttccttaatt ctctgctggc tgataatcat cacctgcagg ttggctccaa   2507 

ttatttgtat attcataaaa tcgataagct ttaatgcggt agtttatcac agttaaattg   2567 

ctaacgcagt caggcaccgt gtatgaaatc taacaatgcg ctcatcgtca tcctcggcac   2627 

cgtcaccctg gatgctgtag gcataggctt ggttatgccg gtactgccgg gcctcttgcg   2687 

ggatcgacgc gaggctggat ggccttcccc attatgattc ttctcgcttc cggcggcatc   2747 

gggatgcccg cgttgcaggc catgctgtcc aggcaggtag atgacgacca tcagggacag   2807 

cttcaaggat cgctcgcggc tcttaccagc ctaacttcga tcactggacc gctgatcgtc   2867 

acggcgattt atgccgcctc ggcgagcaca tggaacgggt tggcatggat tgtaggcgcc   2927 

gccctatacc ttgtctgcct ccccgcgttg cgtcgcggtg catggagccg ggccacctcg   2987 

acctgaatgg aagccggcgg cacctcgcta acggattcac cactccaaga attggagcca   3047 

atcaattctt gcggagaact gtgaatgcgc aaaccaaccc ttggcagaac atatccatcg   3107 

cgtccgccat ctccagcagc cgcacgcggc gcatctcggg cagcgttggg tcctggccac   3167 

gggtgcgcat gatcgtgctc ctgtcgttga ggacccggct aggctggcgg ggttgcctta   3227 

ctggttagca gaatgaatca ccgatacgcg agcgaacgtg aagcgactgc tgctgcaaaa   3287 

cgtctgcgac ctgagcaaca acatgaatgg tcttcggttt ccgtgtttcg taaagtctgg   3347 

aaacgcggaa gtcagcgccc tgcaccatta tgttccggat ctgcatcgca ggatgctgct   3407 

ggctaccctg tggaacacct acatctgtat taacgaagcg ctggcattga ccctgagtga   3467 

tttttctctg gtcccgccgc atccataccg ccagttgttt accctcacaa cgttccagta   3527 

accgggcatg ttcatcatca gtaacccgta tcgtgagcat cctctctcgt ttcatcggta   3587 

tcattacccc catgaacaga aattccccct tacacggagg catcaagtga ccaaacagga   3647 

aaaaaccgcc cttaacatgg cccgctttat cagaagccag acattaacgc ttctggagaa   3707 

actcaacgag ctggacgcgg atgaacaggc agacatctgt gaatcgcttc acgaccacgc   3767 

tgatgagctt taccgcagct gcctcgcgcg tttcggtgat gacggtgaaa acctctgaca   3827 

catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc   3887 

ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc gcagccatga cccagtcacg   3947 

tagcgatagc ggagtgtata ctggcttaac tatgcggcat cagagcagat tgtactgaga   4007 

gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg   4067 

cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg   4127 

gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga   4187 

aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg   4247 

gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag   4307 

aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc   4367 

gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg   4427 

ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt   4487 

cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc   4547 

ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc   4607 

actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg   4667 

tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca   4727 

gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc   4787 

ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat   4847 

cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt   4907 

ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaccccg gttgataatc   4967 

agaaaagccc caaaaacagg aagattgtat aagcaaatat ttaaattgta aacgttaata   5027 

ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg   5087 

aaatcggcaa aatcccttat aaatcaaaag aatagcccga gatagggttg agtgttgttc   5147 

cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa   5207 

ccgtctatca gggcgatggc ccactacgtg aaccatcacc caaatcaagt tttttggggt   5267 

cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac   5327 

ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta   5387 

gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg   5447 

cgccgctaca gggcgcgtaa atcaatctaa agtatatatg agtaaacttg gtctgacagt   5507 

taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata   5567 

gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc   5627 

agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac   5687 

cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag   5747 

tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac   5807 

gttgttgcca ttgctgcagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc   5867 

agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg   5927 

gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc   5987 

atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct   6047 

gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc   6107 

tcttgcccgg cgtcaacacg ggataatacc gcgccacata gcagaacttt aaaagtgctc   6167 

atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc   6227 

agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc   6287 

gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca   6347 

cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt   6407 

tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt   6467 

ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca   6527 

ttaacctata aaaataggcg tatcacgagg ccctttcgtc ttcaagaatt gatcgatcaa   6587 

 
           
             9  
             711  
             DNA  
             Artificial Sequence  
             
               CDS  
               (1)...(708)  
               Description of Artificial Sequence/note =
      synthetic construct  
             
           
            9 

agc aca aaa aag aaa cca tta aca caa gag cag ctt gag gac gca cgt       48 
Ser Thr Lys Lys Lys Pro Leu Thr Gln Glu Gln Leu Glu Asp Ala Arg 
 1               5                   10                  15 

cgc ctt aaa gca att tat gaa aaa aag aaa aat gaa ctt ggc tta tcc       96 
Arg Leu Lys Ala Ile Tyr Glu Lys Lys Lys Asn Glu Leu Gly Leu Ser 
             20                  25                  30 

cag gaa tct gtc gca gac aag atg ggg atg ggg cag tca ggc gtt ggt      144 
Gln Glu Ser Val Ala Asp Lys Met Gly Met Gly Gln Ser Gly Val Gly 
         35                  40                  45 

gct tta ttt aat ggc atc aat gca tta aat gct tat aac gcg gca ttg      192 
Ala Leu Phe Asn Gly Ile Asn Ala Leu Asn Ala Tyr Asn Ala Ala Leu 
     50                  55                  60 

cta gca aaa att ctc aaa gtt agc gtt gaa gaa ttc agc cct tca atc      240 
Leu Ala Lys Ile Leu Lys Val Ser Val Glu Glu Phe Ser Pro Ser Ile 
 65                  70                  75                  80 

gct cga gaa atc tac gag atg tat gaa gcg gtt agc atg cag ccg tca      288 
Ala Arg Glu Ile Tyr Glu Met Tyr Glu Ala Val Ser Met Gln Pro Ser 
                 85                  90                  95 

ctt aga agt gag tat gag tac cct gtt ttt tct cat gtt cag gca ggg      336 
Leu Arg Ser Glu Tyr Glu Tyr Pro Val Phe Ser His Val Gln Ala Gly 
            100                 105                 110 

atg ttc tca cct aag ctt aga acc ttt acc aaa ggt gat gcg gag aga      384 
Met Phe Ser Pro Lys Leu Arg Thr Phe Thr Lys Gly Asp Ala Glu Arg 
        115                 120                 125 

tgg gta agc aca acc aaa aaa gcc agt gat tct gca ttc tgg ctt gag      432 
Trp Val Ser Thr Thr Lys Lys Ala Ser Asp Ser Ala Phe Trp Leu Glu 
    130                 135                 140 

gtt gaa ggt aat tcc atg acc gca cca aca ggc tcc aag cca agc ttt      480 
Val Glu Gly Asn Ser Met Thr Ala Pro Thr Gly Ser Lys Pro Ser Phe 
145                 150                 155                 160 

cct gac gga atg tta att ctc gtt gac cct gag cag gct gtt gag cca      528 
Pro Asp Gly Met Leu Ile Leu Val Asp Pro Glu Gln Ala Val Glu Pro 
                165                 170                 175 

ggt gat ttc tgc ata gcc aga ctt ggg ggt gat gag ttt acc ttc aag      576 
Gly Asp Phe Cys Ile Ala Arg Leu Gly Gly Asp Glu Phe Thr Phe Lys 
            180                 185                 190 

aaa ctg atc agg gat agc ggt cag gtg ttt tta caa cca cta aac cca      624 
Lys Leu Ile Arg Asp Ser Gly Gln Val Phe Leu Gln Pro Leu Asn Pro 
        195                 200                 205 

cag tac cca atg atc cca tgc aat gag agt tgt tcc gtt gtg ggg aaa      672 
Gln Tyr Pro Met Ile Pro Cys Asn Glu Ser Cys Ser Val Val Gly Lys 
    210                 215                 220 

gtt atc gct agt cag tgg cct gaa gag acg ttt ggc tga                  711 
Val Ile Ala Ser Gln Trp Pro Glu Glu Thr Phe Gly 
225                 230                 235 

 
           
             10  
             276  
             DNA  
             Artificial Sequence  
             
               CDS  
               (1)...(276)  
               Description of Artificial Sequence/note =
      synthetic construct  
             
           
            10 

agc aca aaa aag aaa cca tta aca caa gag cag ctt gag gac gca cgt       48 
Ser Thr Lys Lys Lys Pro Leu Thr Gln Glu Gln Leu Glu Asp Ala Arg 
 1               5                   10                  15 

cgc ctt aaa gca att tat gaa aaa aag aaa aat gaa ctt ggc tta tcc       96 
Arg Leu Lys Ala Ile Tyr Glu Lys Lys Lys Asn Glu Leu Gly Leu Ser 
             20                  25                  30 

cag gaa tct gtc gca gac aag atg ggg atg ggg cag tca ggc gtt ggt      144 
Gln Glu Ser Val Ala Asp Lys Met Gly Met Gly Gln Ser Gly Val Gly 
         35                  40                  45 

gct tta ttt aat ggc atc aat gca tta aat gct tat aac gcg gca ttg      192 
Ala Leu Phe Asn Gly Ile Asn Ala Leu Asn Ala Tyr Asn Ala Ala Leu 
     50                  55                  60 

cta gca aaa att ctc aaa gtt agc gtt gaa gaa ttc agc cct tca atc      240 
Leu Ala Lys Ile Leu Lys Val Ser Val Glu Glu Phe Ser Pro Ser Ile 
 65                  70                  75                  80 

gct cga gaa atc tac gag atg tat gaa gcg gtt agc                      276 
Ala Arg Glu Ile Tyr Glu Met Tyr Glu Ala Val Ser 
                 85                  90 

 
           
             11  
             228  
             DNA  
             Artificial Sequence  
             
               CDS  
               (1)...(228)  
               Description of Artificial Sequence/note =
      synthetic construct  
             
           
            11 

agc aca aaa aag aaa cca tta aca caa gag cag ctt gag gac gca cgt       48 
Ser Thr Lys Lys Lys Pro Leu Thr Gln Glu Gln Leu Glu Asp Ala Arg 
 1               5                   10                  15 

cgc ctt aaa gca att tat gaa aaa aag aaa aat gaa ctt ggc tta tcc       96 
Arg Leu Lys Ala Ile Tyr Glu Lys Lys Lys Asn Glu Leu Gly Leu Ser 
             20                  25                  30 

cag gaa tct gtc gca gac aag atg ggg atg ggg cag tca ggc gtt ggt      144 
Gln Glu Ser Val Ala Asp Lys Met Gly Met Gly Gln Ser Gly Val Gly 
         35                  40                  45 

gct tta ttt aat ggc atc aat gca tta aat gct tat aac gcg gca ttg      192 
Ala Leu Phe Asn Gly Ile Asn Ala Leu Asn Ala Tyr Asn Ala Ala Leu 
     50                  55                  60 

cta gca aaa att ctc aaa gtt agc gtt gaa gaa ttc                      228 
Leu Ala Lys Ile Leu Lys Val Ser Val Glu Glu Phe 
 65                  70                  75 

 
           
             12  
             108  
             DNA  
             Artificial Sequence  
             
               CDS  
               (1)...(108)  
               Description of Artificial Sequence/note =
      synthetic construct  
             
           
            12 

agc aca aaa aag aaa cca tta aca caa gag cag ctt gag gac gca cgt       48 
Ser Thr Lys Lys Lys Pro Leu Thr Gln Glu Gln Leu Glu Asp Ala Arg 
 1               5                   10                  15 

cgc ctt aaa gca att tat gaa aaa aag aaa aat gaa ctt ggc tta tcc       96 
Arg Leu Lys Ala Ile Tyr Glu Lys Lys Lys Asn Glu Leu Gly Leu Ser 
             20                  25                  30 

cag gaa tct gtc                                                      108 
Gln Glu Ser Val 
         35 

 
           
             13  
             66  
             DNA  
             Artificial Sequence  
             
               CDS  
               (1)...(66)  
               Description of Artificial Sequence/note =
      synthetic construct  
             
           
            13 

agc aca aaa aag aaa cca tta aca caa gag cag ctt gag gac gca cgt       48 
Ser Thr Lys Lys Lys Pro Leu Thr Gln Glu Gln Leu Glu Asp Ala Arg 
 1               5                   10                  15 

cgc ctt aaa gca att tat                                               66 
Arg Leu Lys Ala Ile Tyr 
             20 

 
           
             14  
             45  
             DNA  
             Artificial Sequence  
             
               CDS  
               (1)...(45)  
               Description of Artificial Sequence/note =
      synthetic construct  
             
           
            14 

agc aca aaa aag aaa cca tta aca caa gag cag ctt gag gac gca           45 
Ser Thr Lys Lys Lys Pro Leu Thr Gln Glu Gln Leu Glu Asp Ala 
 1               5                   10                  15 

 
           
             15  
             6  
             PRT  
             Artificial Sequence  
             
               Description of Artificial Sequence/note =
      synthetic construct  
             
           
            15 

Leu Val Pro Arg Gly Ser 
 1               5 

 
           
             16  
             13  
             DNA  
             Artificial Sequence  
             
               Description of Artificial Sequence/note =
      synthetic construct  
             
           
            16 

aattcgcagc tca                                                        13 

 
           
             17  
             11  
             DNA  
             Artificial Sequence  
             
               Description of Artificial Sequence/note =
      synthetic construct  
             
           
            17 

tatgagctgc g                                                          11 

 
           
             18  
             18  
             DNA  
             Artificial Sequence  
             
               Description of Artificial Sequence/note =
      synthetic construct  
             
           
            18 

acatcagttg agcactgc                                                   18 

 
           
             19  
             27  
             DNA  
             Artificial Sequence  
             
               Description of Artificial Sequence/note =
      synthetic construct  
             
           
            19 

cctcgaggct tgctgataag gacgagc                                         27 

 
           
             20  
             28  
             DNA  
             Artificial Sequence  
             
               Description of Artificial Sequence/note =
      synthetic construct  
             
           
            20 

tcgaggcacc atcaccacca ccactgaa                                        28  
           
             21  
             28  
             DNA  
             Artificial Sequence  
             
               Description of Artificial Sequence/note =
      synthetic construct  
             
           
            21 

agctttcagt ggtggtggtg atggtgcc                                        28 

 
           
             22  
             18  
             DNA  
             Artificial Sequence  
             
               Description of Artificial Sequence/note =
      synthetic construct  
             
           
            22 

tggctggcag tgctcgcc                                                   18 

 
           
             23  
             30  
             DNA  
             Artificial Sequence  
             
               Description of Artificial Sequence/note =
      synthetic construct  
             
           
            23 

tcacctcgag gcctggcaaa acttccgaag                                      30 

 
           
             24  
             26  
             DNA  
             Artificial Sequence  
             
               Description of Artificial Sequence/note =
      synthetic construct  
             
           
            24 

tcgaacggat ccgcgtggca ccaggc                                          26 

 
           
             25  
             18  
             DNA  
             Artificial Sequence  
             
               Description of Artificial Sequence/note =
      synthetic construct  
             
           
            25 

agcgctacct ctcgatcg                                                   18 

 
           
             26  
             32  
             DNA  
             Artificial Sequence  
             
               Description of Artificial Sequence/note =
      synthetic construct  
             
           
            26 

gccgcactcg aggcaaggtc agcctgttta ct                                   32 

 
           
             27  
             40  
             DNA  
             Artificial Sequence  
             
               Description of Artificial Sequence/note =
      synthetic construct  
             
           
            27 

tcgagccacc accaccacca ctctagactg gtgccacgcg                           40 

 
           
             28  
             43  
             DNA  
             Artificial Sequence  
             
               Description of Artificial Sequence/note =
      synthetic construct  
             
           
            28 

gatccgcgtg gcaccagtct agagtggtgg tggtggtggt ggc                       43 

 
           
             29  
             31  
             DNA  
             Artificial Sequence  
             
               Description of Artificial Sequence/note =
      synthetic construct  
             
           
            29 

gcgccatatg gattataagt gtcaagtcca a                                    31 

 
           
             30  
             32  
             DNA  
             Artificial Sequence  
             
               Description of Artificial Sequence/note =
      synthetic construct  
             
           
            30 

gccgctcgag gccaagccca cagatatttc ct                                   32 

 
           
             31  
             39  
             DNA  
             Artificial Sequence  
             
               Description of Artificial Sequence/note =
      synthetic construct  
             
           
            31 

gcgcgaattc accatggaaa tgagacctgc tgtgacttc                            39 

 
           
             32  
             39  
             DNA  
             Artificial Sequence  
             
               Description of Artificial Sequence/note =
      synthetic construct  
             
           
            32 

ccgggctcga ggctagcagt gagtcatttg tactacaat                            39 

 
           
             33  
             35  
             DNA  
             Artificial Sequence  
             
               Description of Artificial Sequence/note =
      synthetic construct  
             
           
            33 

gggaattcca tatgttcaaa cacctccgaa gatgg                                35 

 
           
             34  
             35  
             DNA  
             Artificial Sequence  
             
               Description of Artificial Sequence/note =
      synthetic construct  
             
           
            34 

ccgctcgagg ccatctgggt gtcgtccgtt tcatc                                35 

 
           
             35  
             26  
             DNA  
             Artificial Sequence  
             
               Description of Artificial Sequence/note =
      synthetic construct  
             
           
            35 

gcgcgcatat ggctgccatc cggaag                                          26 

 
           
             36  
             30  
             DNA  
             Artificial Sequence  
             
               Description of Artificial Sequence/note =
      synthetic construct  
             
           
            36 

gccgctcgag gccaagacaa ggcaaccaga                                      30 

 
           
             37  
             26  
             DNA  
             Artificial Sequence  
             
               Description of Artificial Sequence/note =
      synthetic construct  
             
           
            37 

tcgagcctgg tgccacgcgg atccgt                                          26