Patent application title: High Affinity Human Antibodies to Human IL-4 Receptor
Inventors:
Joel H. Martin (Putnam Valley, NY, US)
Tammy T. Huang (Goldens Bridge, NY, US)
Tammy T. Huang (Goldens Bridge, NY, US)
Jeanette L. Fairhurst (White Plains, NY, US)
Jeanette L. Fairhurst (White Plains, NY, US)
Nicholas J. Papadopoulos (Lagrangeville, NY, US)
Assignees:
Regeneron Pharmaceuticals, Inc.
IPC8 Class: AC07K1628FI
USPC Class:
4241721
Class name: Drug, bio-affecting and body treating compositions immunoglobulin, antiserum, antibody, or antibody fragment, except conjugate or complex of the same with nonimmunoglobulin material binds eukaryotic cell or component thereof or substance produced by said eukaryotic cell (e.g., honey, etc.)
Publication date: 2014-09-18
Patent application number: 20140271681
Abstract:
The present invention provides nucleic acid molecules that encode
antibodies or antigen-binding fragments thereof, which specifically bind
human interleukin-4 receptor (IL-4R). Also provided are expression
vectors comprising nucleic acid molecule that encode anti-IL-4R
antibodies, host cells comprising the expression vectors, and methods of
producing anti-IL-4R antibodies or antigen-binding fragments thereof
comprising growing the host cells under conditions permitting production
of the antibody or fragment, and recovering the antibody or fragment so
producedClaims:
1-29. (canceled)
30. An antibody or antigen-binding fragment of an antibody which specifically binds human IL-4R, comprising heavy chain complementarity determining region 1 (HCDR1), 2 (HCDR2), 3 (HCDR3) and light chain complementarity determining region 1 (LCDR1), 2 (LCDR2), 3 (LCDR3), wherein HCDR1 comprises an amino acid sequence of the formula X1-X2-X3-X4-X5-X6-X7-X8 (SEQ ID NO:265), wherein X1=Gly; X2=Phe; X3=Thr; X4=Phe; X5=Asp or Arg; X6=Asp or Ser; X7=Tyr; and X8=Ala or Gly; HCDR2 comprises an amino acid sequence of the formula X1-X2-X3-X4-X5-X6-X7-X8 (SEQ ID NO:266), wherein X1=Ile or Leu, X2=Ser, X3=Gly, Tyr or Arg, X4=Ser, Asp or Thr, X5=Gly or Ser, X6=Gly, Ser or Val, X7=Ser or Asn, and X8=Thr, Lys or Ile; HCDR3 comprises an amino acid sequence of the formula X1-X2-X3-X4-X5-X6-X7-X8-X9-X- 10-X11-X12-X13-X14-X15-X16-X17-X.s- up.18 (SEQ ID NO:267) wherein X1=Ala, X2=Lys, X3=Asp, Glu or Trp, X4=Gly or Arg, X5=Leu, Thr or Arg, X6=Gly, Arg or Ser, X7=Ile or Gly, X8=Thr, Phe or Tyr, X9=Ile, Asp or Phe, X10=Arg, Tyr or Asp, X11=Pro, Tyr or absent, X12=Arg or absent, X13=Tyr or absent, X14=Tyr or absent, X15=Gly or absent, X16=Leu or absent, X17=Asp or absent, and X18=Val or absent; LCDR1 comprises an amino acid sequence of the formula X1-X2-X3-X4-X5-X6-X7-X8-X- 9-X10-X11 (SEQ ID NO:268) wherein X=Gln, X2=Asp, Ser or Val, X3=Ile or Leu, X4=Ser, Leu or Asn, X5=Asn, Tyr or Ile, X6=Trp, Ser or Tyr; X7=ile or absent; X8=Gly or absent; X9=Tyr or absent; X10=Asn or absent; and X11=Tyr or absent; LCDR2 comprises an amino acid sequence of the formula X1-X2-X3 (SEQ ID NO:269) wherein X1=Leu, Ala or Val, X2=Ala or Gly, and X3=Ser; and LCDR3 comprises an amino acid sequence of the formula X1-X2-X3-X4-X5-X6-X7-X8-X9 (SEQ ID NO. 270) wherein X1=Gln or Met, X2=Gln, X3=Ala or Tyr, X4=Leu or Asn, X5=Gn or Ser, X6=Thr, Phe or His, X7=Pro, X8=Tyr, Ile or Trp, and X9=Thr.
31. The antibody or antigen-binding fragment of claim 30, wherein the heavy and light chain CDR amino acid sequences (HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, LCDR3) have the amino acid sequences of SEQ ID NO: 148, 150, 152, 156, 158, and 160, respectively.
32. The antibody or antigen-binding fragment of claim 30, wherein the heavy and light chain CDR amino acid sequences (HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, LCDR3) are encoded by nucleotide sequences of SEQ ID NO:147, 149, 151, 155, 157, and 159, respectively.
33. The antibody or antigen-binding fragment according to claim 30, wherein the antibody or antigen binding fragment comprises a heavy chain variable region (HCVR) having an amino acid sequence of SEQ ID NO: 162 and a light chain variable region (LCVR) having an amino acid sequence of SEQ ID NO: 164.
34. The antibody or antigen-binding fragment according to claim 30, characterized by an affinity for hIL-4R (KD) of ≦300 pM.
35. The antibody or antigen-binding fragment according to claim 30, characterized by an affinity for hIL-4R (KD) of ≦100 pM.
36. The antibody or antigen-binding fragment according to claim 30, characterized by an affinity for hIL-4R (KD) of ≦50 pM.
37. The antibody or antigen-binding fragment according to claim 30, wherein the antibody or fragment thereof cross-reacts with monkey IL-4R.
38. The antibody or antigen binding fragment according to claim 30 for use in the treatment of asthma or atopic dermatitis.
39. A nucleic acid sequence encoding the antibody or antigen binding fragment of claim 30.
40. A vector comprising the nucleic acid sequence of claim 39.
41. An isolated host cell comprising the vector of claim 40.
42. A therapeutic composition comprising the antibody or antigen-binding fragment according to claim 30 and an acceptable carrier.
43. The therapeutic composition of claim 42, further comprising a second therapeutic agent selected from montelukast, pranlukast, zafirlust, and rilonacept.
44. The antibody or antigen binding fragment according to claim 30 for use in the treatment of asthma.
45. The antibody or antigen binding fragment according to claim 30 for use in the treatment of atopic dermatitis.
46. The antibody or antigen binding fragment of claim 30, wherein the antibody or antigen-binding fragment thereof is a human antibody, and wherein the antibody or antigen-binding fragment thereof is made in a transgenic mouse.
47. The antibody or antigen binding fragment of claim 30, wherein the antibody or antigen-binding fragment is a human antibody, and wherein the antibody or antigen-binding fragment thereof is isolated from a hybridoma that is derived from a B cell that is isolated from a transgenic mouse.
48. A therapeutic composition comprising an antibody or antigen-binding fragment, and a pharmaceutically acceptable carrier, wherein the antibody or antigen binding fragment comprises a heavy chain variable region (HCVR) having an amino acid sequence of SEQ ID NO: 162 and a light chain variable region (LCVR) having an amino acid sequence of SEQ ID NO:164.
49. A therapeutic composition comprising an antibody or antigen-binding fragment, and a pharmaceutically acceptable carrier, wherein the antibody or antigen binding fragment comprises heavy and light chain CDR amino acid sequences (HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, LCDR3) having the amino acid sequences of SEQ ID NO:148, 150, 152, 156, 158, and 160, respectively.
Description:
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application is a divisional of U.S. patent application Ser. No. 13/287,151, filed on Nov. 2, 2011, which is a divisional of U.S. patent application Ser. No. 12/556,605, filed on Sep. 10, 2009, issued as U.S. Pat. No. 8,075,887, which is a divisional of U.S. patent application Ser. No. 12/260,307, filed on Oct. 29, 2008, issued as U.S. Pat. No. 7,608,693, which is a continuation-in-part of U.S. patent application Ser. No. 11/906,517, filed on Oct. 2, 2007, issued as U.S. Pat. No. 7,605,237, which claims the benefit under 35 USC §119(e) of U.S. Provisional Application No. 60/848,694, filed on Oct. 2, 2006, and U.S. Provisional Application No. 60/957,738, filed on Aug. 24, 2007, the contents of which are herein specifically incorporated by reference in their entireties.
BACKGROUND
[0002] Interleukin-4 (IL-4, also known as B cell stimulating factor or BSF-1) was originally characterized by its ability to stimulate the proliferation of B cells in response to low concentrations of antibodies directed to surface immunoglobulin. IL-4 has been shown to possess a broad spectrum of biological activities, including growth stimulation of T cells, mast cells, granulocytes, megakaryocytes and erythrocytes. IL-4 induces the expression of class II major histocompatibility complex molecules in resting B cells, and enhances the secretion of IgE and IgG1 isotypes by stimulated B cells.
[0003] The biological activities of IL-4 are mediated by specific cell surface receptors for IL-4. Human IL-4 receptor alpha (hIL-4R) (SEQ ID NO:274) is described in, for example, U.S. Pat. Nos. 5,599,905, 5,767,065, and 5,840,869. Antibodies to hIL-4R are described in U.S. Pat. Nos. 5,717,072 and 7,186,809.
[0004] Methods to produce antibodies useful as human therapeutics include generating chimeric antibodies and humanized antibodies (see, for example, U.S. Pat. No. 6,949,245). See, for example, WO 94/02602 and U.S. Pat. No. 6,596,541 (both of which publications are herein specifically incorporated by reference) describing methods of generating nonhuman transgenic mice capable of producing human antibodies.
[0005] Methods for using antibodies to hIL-4R are described in U.S. Pat. Nos. 5,714,146; 5,985,280; and 6,716,587.
BRIEF SUMMARY OF THE INVENTION
[0006] In a first aspect, the invention provides human antibodies, preferably recombinant human antibodies, that specifically bind human interleukin-4 receptor (hIL-4R). The human antibodies are characterized by binding to hIL-4R with high affinity and by the ability to neutralize hIL-4 activity. In specific embodiments, the human antibodies are capable of blocking hIL-13/hIL-13R1 complex binding to hIL-4R, and thus inhibit signaling by hIL-13. The antibodies can be full-length (for example, an IgG1 or IgG4 antibody) or may comprise only an antigen-binding portion (for example, a Fab, F(ab')2 or scFv fragment), and may be modified to effect functionality, e.g., to eliminate residual effector functions (Reddy et al. (2000) J. Immunol. 164:1925-1933).
[0007] In a general embodiment, the invention provides an antibody or antigen-binding fragment thereof, that specifically binds hIL-4R (SEQ ID NO:274) with a KD of about 300 pM or less, as measured by surface plasmon resonance in a monomeric or dimeric assay. In a more specific embodiment, the antibody or antigen-binding portion thereof exhibits a KD of about 200 pM or less, about 150 or less, about 100 pM or less, or about 50 pM. In various embodiments, the antibody or antigen-binding fragment blocks hIL-4 activity with an IC50 of about 100 pM or less, as measured by luciferase bioassay. In more specific embodiments, the antibody or antigen-binding fragment exhibits an IC50 of about 50 pM or less, about 30 pM or less, or about 25 pM or less, as measured by STAT6 luciferase bioassay. In various embodiments, the antibody or antigen-binding fragment blocks hIL-13 activity with an IC50 of about 100 pM or less, about 90 pM or less, about 50 pM or less, or about 20 pM or less, as measured by STAT6 luciferase bioassay.
[0008] In a second aspect, the antibody of the invention comprises a heavy chain variable region (HCVR) sequence selected from the group consisting of SEQ ID NO:2, 18, 22, 26, 42, 46, 50, 66, 70, 74, 90, 94, 98, 114, 118, 122, 138, 142, 146, 162, 166, 170, 186, 190, 194, 210, 214, 218, 234, 238, 242, 258 and 262, or a substantially similar sequence thereof.
[0009] In a third aspect, the antibody of the invention comprises a light chain variable region (LCVR) sequence selected from the group consisting of SEQ ID NO:10, 20, 24, 34, 44, 48, 58, 68, 72, 82, 92, 96, 106, 116, 120, 130, 140, 144, 154, 164, 168, 178, 188, 192, 202, 212, 216, 226, 236, 240, 250, 260 and 264, or a substantially similar sequence thereof.
[0010] In one embodiment, the antibody or antibody fragment of the invention comprises HCVR and LCVR sequence pairs (HCVR/LCVR) selected from the group consisting of SEQ ID NO: 2/10, 18/20, 22/24, 26/34, 42/44, 46/48, 50/58, 66/68, 70/72, 74/82, 90/92, 94/96, 98/106, 114/116, 118/120, 122/130, 138/140, 142/144, 146/154, 162/164, 166/168, 170/178, 186/188, 190/192, 194/202, 210/212, 214/216, 218/226, 234/236, 238/240, 242/250, 258/260 and 262/264. In a preferred embodiment, the antibody or antibody fragment comprise HCVR/LCVR sequence pairs SEQ ID NO:162/164, 210/212 and 18/20. In an even more preferred embodiment, the antibody or antibody fragment comprises HCVR/LCVR SEQ ID NO:162/164.
[0011] In a fourth aspect, the invention provides nucleic acid molecules encoding an HCVR, wherein the nucleic acid molecule is a nucleotide sequence selected from the group consisting of SEQ ID NO: 1, 17, 21, 25, 41, 45, 49, 65, 69, 73, 89, 93, 97, 113, 117, 121, 137, 141, 145, 161, 165, 169, 185, 189, 193, 209, 213, 217, 233, 237, 241, 257 and 261, or a substantially identical sequence having at least 95% homology thereof.
[0012] In a fifth aspect, the invention provides nucleic acid molecules encoding a LCVR, wherein the nucleic acid molecule is a sequence selected from the group consisting of SEQ ID NO: 9, 19, 23, 33, 43, 47, 57, 67, 71, 81, 91, 95, 105, 115, 119, 129, 139, 143, 153, 163, 167, 177, 187, 191, 201, 211, 215, 225, 235, 239, 249, 259 and 263, or a substantially identical sequence having at least 95% homology thereof.
[0013] In one embodiment, the antibody of the invention comprises a HCVR and LCVR encoded by a nucleotide sequence pairs selected from the group consisting of SEQ ID NO: 1/9, 17/19, 21/22, 25/33, 41/43, 45/47, 49/57, 65/67, 69/71, 73/81, 89/91, 93/95, 97/105, 113/115, 117/119, 121/129, 137/139, 141/143, 145/153, 161/163, 165/167, 169/177, 185/187, 189/191, 193/201, 209/211, 213/215, 217/225, 233/235, 237/239, 241/249, 257/259 and 261/263. In a preferred embodiment, the antibody or antibody fragment comprise HCVR/LCVR sequences encoded by nucleic acid sequences selected from SEQ ID NO:161/163, 209/211 and 17/19. In an even more preferred embodiment, the antibody or antibody fragment comprises HCVR/LCVR encoded by nucleic acid sequences SEQ ID NO:161/163.
[0014] In a sixth aspect, the invention provides an antibody or antigen-binding fragment comprising a HCDR3 and a LCDR3, wherein the HCDR3 domain selected from the group consisting of SEQ ID NO:8, 32, 56, 80, 104, 128, 152, 176, 200, 224 and 248; and the LCDR3 domain selected from the group consisting of SEQ ID NO:16, 40, 64, 88, 112, 136, 160, 184, 208, 232 and 256. In a preferred embodiment, the HCDR3/LCDR3 sequences are SEQ ID NO:152/160, 8/16 or 200/208. In an even more preferred embodiment, the HCDR3 and LCDR3 sequences are SEQ ID NO:152 and 160.
[0015] In a further embodiment, the antibody or antibody fragment further comprises a HCDR1 sequence selected from the group consisting of SEQ ID NO:4, 28, 52, 76, 100, 124, 148, 172, 196, 220 and 244, or a substantially similar sequence thereof; a HCDR2 sequence selected from the group consisting of SEQ ID NO:6, 30, 54, 78, 102, 126, 150, 174, 198, 222 and 246, or a substantially similar sequence thereof; a HCDR3 sequence selected from the group consisting of SEQ ID NO:8, 32, 56, 80, 104, 128, 152, 176, 200, 224 and 248, or a substantially similar sequence thereof; a LCDR1 sequence selected from the group consisting of SEQ ID NO:12, 36, 60, 84, 108, 132, 156, 180, 204, 228 and 252, or a substantially similar sequence thereof; a LCDR2 sequence selected from the group consisting of SEQ ID NO:14, 38, 62, 86, 110, 134, 158, 182, 206, 230 and 252, or a substantially similar sequence thereof; and a LCDR3 sequence selected from the group consisting of SEQ ID NO:16, 40, 64, 88, 112, 136, 160, 184, 208, 232 and 256 or a substantially similar sequences thereof. In a preferred embodiment, the antibody or antigen-binding fragment comprise HCDR sequences SEQ ID NO:148, 150 and 152 and LCDR sequences SEQ ID NO:156, 158 and 160; HCDR sequences SEQ ID NO:4, 6 and 8 and LCDR sequences SEQ ID NO:12, 14 and 16; and HCDR sequences SEQ ID NO:196, 198 and 200 and LCDR sequences SEQ ID NO:204, 206 and 208.
[0016] In a seventh aspect, the invention features a human antibody or antibody fragment comprising a HCDR3 and LCDR3, wherein the HCDR3 is encoded by a nucleotide sequence selected from the group consisting of SEQ ID NO:7, 31, 55, 79, 103, 127, 151, 175, 199, 223 and 247, or a substantially identical sequence having at least 95% homology thereof; and the LCDR3 is encoded by a nucleotide sequence selected from the group consisting of SEQ ID NO:15, 39, 63, 87, 111, 135, 159, 183, 207, 231 and 255, or a substantially identical sequence having at least 95% homology thereof.
[0017] In a further embodiment, the invention features a human antibody or antibody fragment comprising a HCDR1 domain encoded by a nucleotide sequence selected from the group consisting of SEQ ID NO:3, 27, 51, 75, 99, 123, 147, 171, 195, 219 and 243, or a substantially identical sequence having at least 95% homology thereof; a HCDR2 domain encoded by a nucleotide sequence selected from the group consisting of SEQ ID NO:5, 29, 53, 77, 101, 125, 149, 173, 197, 221 and 245, or a substantially identical sequence having at least 95% homology thereof; a HCDR3 domain encoded by a nucleotide sequence selected from the group consisting of SEQ ID NO: 7, 31, 55, 79, 103, 127, 151, 175, 199, 223 and 247, or a substantially similar sequence having at least 95% homology thereof; a LCDR1 domain encoded by a nucleotide sequence selected from the group consisting of SEQ ID NO:11, 35, 59, 83, 107, 131, 155, 179, 203, 227 and 251, or a substantially similar sequence having at least 95% homology thereof; a LCDR2 domain encoded by a nucleotide sequence selected from the group consisting of SEQ ID NO:13, 37, 61, 85, 109, 133, 157, 181, 205, 229 and 253, or a substantially similar sequence having at least 95% homology thereof; and LCDR3 domain encoded by a nucleotide sequence selected from the group consisting of SEQ ID NO: 15, 39, 63, 87, 111, 135, 159, 183, 207, 231 and 255, or a substantially similar sequence having at least 95% homology thereof. In a preferred embodiment, the antibody or antigen-binding fragment comprise HCDR and LCDR sequences encoded by nucleotide sequences SEQ ID NO:147, 149, 151, 155, 157 and 159; 195, 197, 199, 203, 205 and 207; and 3, 5, 7, 11, 13 and 15.
[0018] In a specific embodiment, the anti-hIL-4R antibody or antigen-binding fragment of the invention comprises HCVR comprising the amino acid sequence shown in SEQ ID NO:162 and LCVR comprising the amino acid sequence shown in SEQ ID NO:164, and is characterized by a KD of about 100 pM or less (monomeric substrate) or 70 pM or less (dimeric substrate); a KD of about 160 pM or less (monomeric substrate) or 40 pM or less (dimeric substrate) at 25° C. and 37° C., respectively; and an IC50 of about 10 pM or less (25 pM dimer substrate) or about 100 pM or less (200 pM monomer substrate), which is capable of blocking both hIL-4 and hIL-13 activity with an IC50 of about 30 pM or less (as measured by bioassay) and cross-reacts with monkey IL-4R.
[0019] In a specific embodiment, the anti-hIL-4R antibody or antigen-binding fragment of the invention comprises HCVR comprising the amino acid sequence shown in SEQ ID NO:18 and LCVR comprising the amino acid sequence shown in SEQ ID NO:20, and is characterized by a KD of about 450 pM or less (monomeric or dimeric substrate); and an IC50 of about 40 pM or less (25 pM dimer substrate) or about 100 pM or less (200 pM monomer substrate), which is capable of blocking both hIL-4 and hIL-13 activity with an IC50 of about 100 pM or less (as measured by bioassay).
[0020] In a specific embodiment, the anti-hIL-4R antibody or antigen-binding fragment of the invention comprises HCVR comprising the amino acid sequence shown in SEQ ID NO:210 and LCVR comprising the amino acid sequence shown in SEQ ID NO:212, and is characterized by a KD of about 50 pM or less (monomeric substrate) or 30 pM or less (dimeric substrate); a KD of about 200 pM or less (monomeric substrate) or 40 pM or less (dimeric substrate) at 25° C. and 37° C., respectively; and an IC50 of about 10 pM or less (25 pM dimer substrate) or about 90 pM or less (200 pM monomer substrate), which is capable of blocking both hIL-4 and hIL-13 activity with an IC50 of about 25 pM or less (as measured by bioassay) and does not cross-reacts with monkey IL-4R.
[0021] In a eighth aspect, the invention features an antibody or antigen-binding fragment of an antibody that specifically binds hIL-4R, comprising three heavy chain and three light chain complementarity determining regions (HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, LCDR3), wherein the HCDR1 comprises an amino acid sequence of the formula X1-X2-X3-X4-X5-X6-X7-X8 (SEQ ID NO:265), wherein X1=Gly; X2=Phe; X3=Thr; X4=Phe; X5=Asp or Arg; X6=Asp or Ser; X7=Tyr; and X8=Ala or Gly; the HCDR2 comprises an amino acid sequence of the formula X1-X2-X3-X4-X5-X6-X7-X8 (SEQ ID NO:266), wherein X1=Ile or Leu, X2=Ser, X3=Gly, Tyr or Arg, X4=Ser, Asp or Thr, X5=Gly or Ser, X6=Gly, Ser or Val, X7=Ser or Asn, and X8=Thr, Lys or Ile; the HCDR3 comprises an amino acid sequence of the formula X1-X2-X3-X4-X5-X6-X7-X8-X9-X- 10-X11-X12-X13-X14-X15-X16-X17-X.s- up.18 (SEQ ID NO:267) wherein X1=Ala, X2=Lys, X3=Asp, Glu or Trp, X4=Gly or Arg, X5=Leu, Thr or Arg, X6=Gly, Arg or Ser, X7=Ile or Gly, X8=Thr, Phe or Tyr, X9=Ile, Asp or Phe, X10=Arg, Tyr or Asp, X11=Pro, Tyr or absent, X12=Arg or absent, X13=Tyr or absent, X14=Tyr or absent, X15=Gly or absent, X16=Leu or absent, X17=Asp or absent, and X18=Val or absent; the LCDR1 comprises an amino acid sequence of the formula X1-X2-X3-X4-X5-X6-X7-X8-X- 9-X10-X11 (SEQ ID NO:268) wherein X1=Gln, X2=Asp, Ser or Val, X3=Ile or Leu, X4=Ser, Leu or Asn, X5=Asn, Tyr or Ile, X6=Trp, Ser or Tyr; X7=Ile or absent; X8=Gly or absent; X9=Tyr or absent; X10=Asn or absent; and X11=Tyr or absent; the LCDR2 comprises an amino acid sequence of the formula X1-X2-X3 (SEQ ID NO:269) wherein X1=Leu, Ala or Val, X2=Ala or Gly, and X3=Ser; and the LCDR3 comprises an amino acid sequence of the formula X1-X2-X3-X4-X5-X6-X7-X8-X9 (SEQ ID NO:270) wherein X1=Gln or Met, X2=Gln, X3=Ala or Tyr, X4=Leu or Asn, X5=Gln or Ser, X6=Thr, Phe or His, X7=Pro, X8=Tyr, Ile or Trp, and X9=Thr.
[0022] In a more specific embodiment, the HCDR1 comprises an amino acid sequence of the formula X1-X2-X3-X4-X5-X6-X7-X8 (SEQ ID NO:265), wherein X1=Gly; X2=Phe; X3=Thr; X4=Phe; X5=Arg; X6=Asp or Ser; X7=Tyr; and X8=Ala or Gly; the HCDR2 comprises an amino acid sequence of the formula X1-X2-X3-X4-X5-X6-X7-X8 (SEQ ID NO:266), wherein X1=Ile, X2=Ser, X3=Gly or Tyr, X4=Ser or Thr, X5=Gly, X6=Gly or Ser, X7=Asn, and X8=Thr or Lys; the HCDR3 comprises an amino acid sequence of the formula X1-X2-X3-X4-X5-X6-X7-X8-X- 9-X10-X11-X12-X13-X14-X15-X16-X.su- p.17-X18 (SEQ ID NO:267) wherein X1=Ala, X2=Lys, X3=Asp or Glu, X4=Gly or Arg, X5=Leu or Arg, X6=Gly or Ser, X7=Ile or Gly, X8=Thr or Phe, X9=lie or Asp, X10=Arg or Tyr, X11=Pro or absent, X12=Arg or absent, X13=Tyr or absent, X14=Tyr or absent, X15=Gly or absent, X16=Leu or absent, X17=Asp or absent, and X18=Val or absent; the LCDR1 comprises an amino acid sequence of the formula X1-X2-X3-X4-X5-X6-X7-X8-X9-X- 10-X11 (SEQ ID NO:268) wherein X1=Gln, X2=Ser or Val, X3=Ile or Leu, X4=Leu or Asn, X5=Asn or Tyr, X6=Ser or Tyr; X7=Ile or absent; X8=Gly or absent; X9=Tyr or absent; X10=Asn or absent; and X11=Tyr or absent; the LCDR2 comprises an amino acid sequence of the formula X1-X2-X3 (SEQ ID NO:269) wherein X1=Leu or Ala, X2=Ala or Gly, and X3=Ser; and the LCDR3 comprises an amino acid sequence of the formula X1-X2-X3-X4-X5-X6-X7-X8-X- 9 (SEQ ID NO:270) wherein X1=Gln or Met, X2=Gln, X3=Ala or Tyr, X4=Leu or Asn, X5=Gln or Ser, X6=Thr or His, X7=Pro, X8=Tyr or Trp, and X9=Thr.
[0023] In another more specific embodiment, the HCDR1 comprises an amino acid sequence of the formula X1-X2-X3-X4-X5-X6-X7-X8 (SEQ ID NO:265), wherein X1=Gly; X2=Phe; X3=Thr; X4=Phe; X5=Asp or Arg; X6=Asp; X7=Tyr; and X8=Ala; the HCDR2 comprises an amino acid sequence of the formula X1-X2-X3-X4-X5-X6-X7-X8 (SEQ ID NO:266), wherein X1=Ile or Leu, X2=Ser, X3=Gly or Arg, X4=Ser or Thr, X5=Gly or Ser, X6=Gly or Val, X7=Ser or Asn, and X8=Thr or Ile; the HCDR3 comprises an amino acid sequence of the formula X1-X2-X3-X4-X5-X6-X7-X8-X9-X- 10-X11-X12-X13-X14-X15-X16-X17-X.s- up.18 (SEQ ID NO:267) wherein X1=Ala, X2=Lys, X3=Asp or Trp, X4=Gly or Arg, X5=Leu or Thr, X6=Arg or Ser, X7=Ile or Gly, X8=Thr or Tyr, X9=lie or Phe, X10=Arg or Asp, X11=Pro, Tyr or absent, X12=Arg or absent, X13=Tyr or absent, X14=Tyr or absent, X15=Gly or absent, X16=Leu or absent, X17=Asp or absent, and X18=Val or absent; the LCDR1 comprises an amino acid sequence of the formula X1-X2-X3-X4-X5-X6-X7-X8-X9-X- 10-X11 (SEQ ID NO:268) wherein X1=Gln, X2=Asp or Ser, X3=Ile or Leu, X4=Ser or Leu, X5=Tyr or Ile, X6=Trp or Ser; X7=Ile or absent; X8=Gly or absent; X9=Tyr or absent; X10=Asn or absent; and X11=Tyr or absent; the LCDR2 comprises an amino acid sequence of the formula X1-X2-X3 (SEQ ID NO:269) wherein X1=Leu or Val, X2=Ala or Gly, and X3=Ser; and the LCDR3 comprises an amino acid sequence of the formula X1-X2-X3-X4-X5-X6-X7-X8-X- 9 (SEQ ID NO:270) wherein X1=Gln or Met, X2=Gln, X3=Ala, X4=Leu or Asn, X5=Gln or Ser, X6=Thr or Phe, X7=Pro, X8=Tyr or Ile, and X9=Thr.
[0024] In a ninth aspect, the invention provides an antibody or antigen-binding fragment comprising HCDR1/HCDR2/HCDR3/LCDR1/LCDR2/LCDR3 sequences from a HCVR and LCVR pair, wherein the HCVR/LCVR sequences are selected from the group consisting of SEQ ID NO:162/164, 210/212 and 18/20. In a more specific embodiment, heavy and light chain CDR sequences are those contained in HCVR SEQ ID NO:162 and LCVR SEQ ID NO:164. In another more specific embodiment, heavy and light chain CDR sequences are those contained in HCVR SEQ ID NO:18 and LCVR SEQ ID NO:20. In yet another specific embodiment, heavy and light chain CDR sequences are those contained in HCVR SEQ ID NO:210 and LCVR SEQ ID NO:212.
[0025] The invention encompasses anti-hIL-4R antibodies having a modified glycosylation pattern. In some applications, modification to remove undesirable glycosylation sites may be useful, or an antibody lacking a fucose moiety present on the oligosaccharide chain, for example, to increase antibody dependent cellular cytotoxicity (ADCC) function (see Shield et al. (2002) JBC 277:26733). In other applications, modification of a galactosylation can be made in order to modify complement dependent cytotoxicity (CDC).
[0026] In a tenth aspect, the invention provides recombinant expression vectors carrying the nucleic acid molecules of the invention, and host cells into which such vectors have been included, as are methods of making the antibodies or antigen-binding fragments of the invention obtained by culturing the host cells of the invention. The host cell may be a prokaryotic or eukaryotic cell, preferably the host cell is an E. coli cell or a mammalian cell, such as a CHO cell.
[0027] In an eleventh aspect, the invention features a composition comprising a recombinant human antibody that specifically binds hIL-4R and an acceptable carrier.
[0028] In a twelfth aspect, the invention features methods for inhibiting hIL-4 activity using an antibody, or antigen-binding portion thereof, of the invention. In specific embodiments, the antibodies of the invention also block hIL-13/hIL-13R1 complex binding to hIL-4R. In one embodiment, the method comprises contacting hIL-4R with the antibody of the invention, or antigen-binding portion thereof, such that hIL-4 or hIL-4/hIL-13 activity is inhibited. In another embodiment, the method comprises administering an antibody of the invention, or antigen-binding portion thereof, to a human subject suffering from a disorder that is ameliorated by inhibition of hIL-4 or hIL-4/hIL-13 activity. The disorder treated is any disease or condition that is improved, ameliorated, inhibited or prevented by removal, inhibition or reduction of hIL-4 or hIL-4/hIL-13 activity.
[0029] IL-4 related disorders which are treated by the antibodies or antibody fragments of the invention include, for example, arthritis (including septic arthritis), herpetiformis, chronic idiopathic urticaria, scleroderma, hypertrophic scarring, Whipple's Disease, benign prostate hyperplasia, lung disorders, such as mild, moderate or severe asthma, inflammatory disorders such as inflammatory bowel disease, allergic reactions, Kawasaki disease, sickle cell disease, Churg-Strauss syndrome, Grave's disease, pre-eclampsia, Sjogren's syndrome, autoimmune lymphoproliferative syndrome, autoimmune hemolytic anemia, Barrett's esophagus, autoimmune uveitis, tuberculosis, and nephrosis.
[0030] Other objects and advantages will become apparent from a review of the ensuing detailed description.
DETAILED DESCRIPTION
[0031] Before the present methods are described, it is to be understood that this invention is not limited to particular methods, and experimental conditions described, as such methods and conditions may vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting, since the scope of the present invention will be limited only by the appended claims.
[0032] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, the preferred methods and materials are now described. All publications mentioned herein are incorporated herein by reference in their entirety.
DEFINITIONS
[0033] The term "human IL4R" (hIL-4R), as used herein, is intended to refer to a human cytokine receptor that specifically binds interleukin-4 (IL-4), IL-4Rα (SEQ ID NO:274). The term "human interleukin-13" (hIL-13) refers to a cytokine that specifically binds IL-13 receptor, and "hIL-13/hIL-13R1 complex" refers to the complex formed by hIL-13 binding to hIL-13R1 complex, which complex binds hIL-4 receptor to initiate biological activity.
[0034] The term "antibody", as used herein, is intended to refer to immunoglobulin molecules comprising four polypeptide chains, two heavy (H) chains and two light (L) chains inter-connected by disulfide bonds. Each heavy chain comprises a heavy chain variable region (HCVR or VH) and a heavy chain constant region. The heavy chain constant region comprises three domains, CH1, CH2 and CH3. Each light chain comprises a light chain variable region (LCVR or VL) and a light chain constant region. The light chain constant region comprises one domain (CL1). The VH and VL regions can be further subdivided into regions of hypervariability, termed complementarity determining regions (CDR), interspersed with regions that are more conserved, termed framework regions (FR). Each VH and VL is composed of three CDRs and four FRs, arranged from amino-terminus to carboxy-terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4.
[0035] The term "antigen-binding portion" of an antibody (or simply "antibody portion" or "antibody fragment"), as used herein, refers to one or more fragments of an antibody that retain the ability to specifically bind to an antigen (e.g., hIL-4R). It has been shown that the antigen-binding function of an antibody can be performed by fragments of a full-length antibody. Examples of binding fragments encompassed within the term "antigen-binding portion" of an antibody include (i) a Fab fragment, a monovalent fragment consisting of the VL, VH, CL1 and CH1 domains; (ii) a F(ab')2 fragment, a bivalent fragment comprising two F(ab)' fragments linked by a disulfide bridge at the hinge region; (iii) a Fd fragment consisting of the VH and CH1 domains; (iv) a Fv fragment consisting of the VL and VH domains of a single arm of an antibody, (v) a dAb fragment (Ward et al. (1989) Nature 241:544-546), which consists of a VH domain; and (vi) an CDR. Furthermore, although the two domains of the Fv fragment, VL and VH, are coded for by separate genes, they can be joined, using recombinant methods, by a synthetic linker that enables them to be made as a single contiguous chain in which the VL and VH regions pair to form monovalent molecules (known as single chain Fv (scFv); see e.g., Bird et al. (1988) Science 242:423-426; and Huston et al. (1988) Proc. Natl. Acad. Sci. USA 85:5879-5883. Such single chain antibodies are also intended to be encompassed within the term "antigen-binding portion" of an antibody. Other forms of single chain antibodies, such as diabodies, are also encompassed (see e.g., Holliger et al. (1993) Proc. Natl. Acad. Sci. USA 90:6444-6448).
[0036] A "neutralizing" or "blocking" antibody, as used herein, is intended to refer to an antibody whose binding to hIL-4R results in inhibition of the biological activity of hIL-4 and/or hIL-13. This inhibition of the biological activity of hIL-4 and/or IL-13 can be assessed by measuring one or more indicators of hIL-4 and/or hIL-13 biological activity known to the art, such as hIL-4- and/or IL-13-induced cellular activation and hIL-4 binding to hIL-4R (see examples below).
[0037] A "CDR" or complementarity determining region is a region of hypervariability interspersed within regions that are more conserved, termed "framework regions" (FR). In different embodiments of the anti-hIL-4R antibody or fragment of the invention, the FRs may be identical to the human germline sequences, or may be naturally or artificially modified.
[0038] The term "surface plasmon resonance", as used herein, refers to an optical phenomenon that allows for the analysis of real-time interactions by detection of alterations in protein concentrations within a biosensor matrix, for example using the BIACORE® system (Pharmacia Biosensor AB).
[0039] The term "epitope" is an antigenic determinant that interacts with a specific antigen binding site in the variable region of an antibody molecule known as a paratope. A single antigen may have more than one epitope. Epitopes may be either conformational or linear. A conformational epitope is produced by spatially juxtaposed amino acids from different segments of the linear polypeptide chain. A linear epitope is one produced by adjacent amino acid residues in a polypeptide chain. In certain circumstance, an epitope may include moieties of saccharides, phosphoryl groups, or sulfonyl groups on the antigen.
[0040] The term "substantial identity" or "substantially identical," when referring to a nucleic acid or fragment thereof, indicates that, when optimally aligned with appropriate nucleotide insertions or deletions with another nucleic acid (or its complementary strand), there is nucleotide sequence identity in at least about 95%, and more preferably at least about 96%, 97%, 98% or 99% of the nucleotide bases, as measured by any well-known algorithm of sequence identity, such as FASTA, BLAST or Gap, as discussed below.
[0041] As applied to polypeptides, the term "substantial similarity" or "substantially similar" means that two peptide sequences, when optimally aligned, such as by the programs GAP or BESTFIT using default gap weights, share at least 95% sequence identity, even more preferably at least 98% or 99% sequence identity. Preferably, residue positions which are not identical differ by conservative amino acid substitutions. A "conservative amino acid substitution" is one in which an amino acid residue is substituted by another amino acid residue having a side chain (R group) with similar chemical properties (e.g., charge or hydrophobicity). In general, a conservative amino acid substitution will not substantially change the functional properties of a protein. In cases where two or more amino acid sequences differ from each other by conservative substitutions, the percent sequence identity or degree of similarity may be adjusted upwards to correct for the conservative nature of the substitution. Means for making this adjustment are well-known to those of skill in the art. See, e.g., Pearson (1994) Methods Mol. Biol. 24: 307-331, herein incorporated by reference. Examples of groups of amino acids that have side chains with similar chemical properties include (1) aliphatic side chains: glycine, alanine, valine, leucine and isoleucine; (2) aliphatic-hydroxyl side chains: serine and threonine; (3) amide-containing side chains: asparagine and glutamine; (4) aromatic side chains: phenylalanine, tyrosine, and tryptophan; (5) basic side chains: lysine, arginine, and histidine; (6) acidic side chains: aspartate and glutamate, and (7) sulfur-containing side chains are cysteine and methionine. Preferred conservative amino acids substitution groups are: valine-leucine-isoleucine, phenylalanine-tyrosine, lysine-arginine, alanine-valine, glutamate-aspartate, and asparagine-glutamine. Alternatively, a conservative replacement is any change having a positive value in the PAM250 log-likelihood matrix disclosed in Gonnet et al. (1992) Science 256: 1443 45, herein incorporated by reference. A "moderately conservative" replacement is any change having a nonnegative value in the PAM250 log-likelihood matrix.
[0042] Sequence similarity for polypeptides, which is also referred to as sequence identity, is typically measured using sequence analysis software. Protein analysis software matches similar sequences using measures of similarity assigned to various substitutions, deletions and other modifications, including conservative amino acid substitutions. For instance, GCG software contains programs such as Gap and Bestfit which can be used with default parameters to determine sequence homology or sequence identity between closely related polypeptides, such as homologous polypeptides from different species of organisms or between a wild type protein and a mutein thereof. See, e.g., GCG Version 6.1. Polypeptide sequences also can be compared using FASTA using default or recommended parameters, a program in GCG Version 6.1. FASTA (e.g., FASTA2 and FASTA3) provides alignments and percent sequence identity of the regions of the best overlap between the query and search sequences (Pearson (2000) supra). Another preferred algorithm when comparing a sequence of the invention to a database containing a large number of sequences from different organisms is the computer program BLAST, especially BLASTP or TBLASTN, using default parameters. See, e.g., Altschul et al. (1990) J. Mol. Biol. 215:403-410 and Altschul et al. (1997) Nucleic Acids Res. 25:3389-402, each of which is herein incorporated by reference.
Preparation of Human Antibodies
[0043] Methods for generating human antibodies include those described in, for example, U.S. Pat. No. 6,596,541, Green et al. (1994) Nature Genetics 7:13-21), U.S. Pat. No. 5,545,807, U.S. Pat. No. 6,787,637.
[0044] Rodents can be immunized by any method known in the art (see, for example, Harlow and Lane (1988) Antibodies: A Laboratory Manual 1988 Cold Spring Harbor Laboratory; Malik and Lillehoj (1994) Antibody Techniques, Academic Press, CA). Antibodies of the invention are preferably prepared with the use of VELOCIMMUNE® technology (U.S. Pat. No. 6,596,541). A transgenic mouse in which the endogenous immunoglobulin heavy and light chain variable regions are replaced with the corresponding human variable regions is challenged with the antigen of interest, and lymphatic cells (such as B-cells) are recovered from the mice that express antibodies. The lymphatic cells may be fused with a myeloma cell line to prepare immortal hybridoma cell lines, and such hybridoma cell lines are screened and selected to identify hybridoma cell lines that produce antibodies specific to the antigen of interest. DNA encoding the variable regions of the heavy chain and light chain may be isolated and linked to desirable isotypic constant regions of the heavy chain and light chain. Such an antibody protein may be produced in a cell, such as a CHO cell. Alternatively, DNA encoding the antigen-specific chimeric antibodies or the variable regions of the light and heavy chains may be isolated directly from antigen-specific lymphocytes.
[0045] The DNA encoding the variable regions of the heavy and light chains of the antibody are isolated and operably linked to DNA encoding the human heavy and light chain constant regions. The DNA is then expressed in a cell capable of expressing the fully human antibody. In a specific embodiment, the cell is a CHO cell.
[0046] Antibodies may be therapeutically useful in blocking a ligand-receptor interaction or inhibiting receptor component interaction, rather than by killing cells through fixation of complement (complement-dependent cytotoxicity) (CDC) and participation antibody-dependent cell-mediated cytotoxicity (ADCC). The constant region of an antibody is important in the ability of an antibody to fix complement and mediate cell-dependent cytotoxicity. Thus, the isotype of an antibody may be selected on the basis of whether it is desirable for the antibody to mediate cytotoxicity.
[0047] Human immunoglobulins can exist in two forms that are associated with hinge heterogeneity. In one form, an immunoglobulin molecule comprises a stable four-chain construct of approximately 150-160 kDa in which the dimers are held together by an interchain heavy chain disulfide bond. In a second form, the dimers are not linked via interchain disulfide bonds and a molecule of about 75-80 kDa is formed composed of a covalently coupled light and heavy chain (half-antibody). These forms have been extremely difficult to separate, even after affinity purification. The frequency of appearance of the second form in various intact IgG isotypes is due to, but not limited to, structural differences associated with the hinge region isotype of the antibody. In fact, a single amino acid substitution in the hinge region of the human IgG4 hinge can significantly reduce the appearance of the second form (Angal et al. (1993) Molecular Immunology 30: 105) to levels typically observed using a human IgG1 hinge. The instant invention encompasses antibodies having one or more mutations in the hinge, CH2 or CH3 region that may be desirable, for example, in production, to improve the yield of the desired antibody form.
[0048] Initially, high affinity chimeric antibodies are isolated having a human variable region and a mouse constant region. As described below, the antibodies are characterized and selected for desirable characteristics, including binding affinity to hIL-4R, ability to block hIL-4 binding to hIL-4R, and/or selectivity for the human protein. The mouse constant regions are replaced with desired human constant regions to generate the fully human antibodies of the invention, for example wild-type or modified IgG4 or IgG1 (for example, SEQ ID NO:271, 272, 273). While the constant region selected may vary according to specific use, high affinity antigen-binding and target specificity characteristics reside in the variable region.
Epitope Mapping and Related Technologies
[0049] To screen for antibodies that bind to a particular epitope, a routine cross-blocking assay such as that described in Harlow and Lane supra can be performed. Other methods include alanine scanning mutants, peptide blots (Reineke (2004) Methods Mol Biol 248:443-63), or peptide cleavage analysis. In addition, methods such as epitope excision, epitope extraction and chemical modification of antigens can be employed (Tomer (2000) Protein Science: 9:487-496).
[0050] Modification-Assisted Profiling (MAP), also known as Antigen Structure-based Antibody Profiling (ASAP) is a method that categorizes large numbers of monoclonal antibodies (mAbs) directed against the same antigen according to the similarities of the binding profile of each antibody to chemically or enzymatically modified antigen surfaces (US Patent Application Publication No. 2004/0101920, herein specifically incorporated by reference in its entirety). Each category may reflect a unique epitope either distinctly different from, or partially overlapping with, an epitope represented by another category. This technology allows rapid filtering of genetically identical antibodies, such that characterization can be focused on genetically distinct antibodies. When applied to hybridoma screening, MAP may facilitate identification of rare hybridoma clones with desired characteristics. MAP may be used to sort the hIL-4R antibodies of the invention into groups of antibodies binding different epitopes.
[0051] Agents useful for altering the structure of the immobilized antigen are enzymes, such as, for example, proteolytic enzymes and chemical agents. The antigen protein may be immobilized on either biosensor chip surfaces or polystyrene beads. The latter can be processed with, for example, an assay such as a multiplex LUMINEX® detection assay (Luminex Corp., TX). Because of the capacity of LUMINEX® to handle multiplex analysis with up to 100 different types of beads, LUMINEX® provides almost unlimited antigen surfaces with various modifications, resulting in improved resolution in antibody epitope profiling over a biosensor assay.
Bispecifics
[0052] The antibodies of the present invention may be monospecific, bispecific, or multispecific. Multispecific antibodies may be specific for different epitopes of one target polypeptide or may contain antigen-binding domains specific for more than one target polypeptide. See, e.g., Tutt et al. (1991) J. Immunol. 147:60-69. The human anti-IL-4R antibodies can be linked to or co-expressed with another functional molecule, e.g., another peptide or protein. For example, an antibody or fragment thereof can be functionally linked (e.g., by chemical coupling, genetic fusion, noncovalent association or otherwise) to one or more other molecular entities, such as another antibody or antibody fragment, to produce a bispecific or a multispecific antibody with a second binding specificity.
Therapeutic Administration and Formulations
[0053] The invention provides therapeutic compositions comprising the anti-IL-4R antibodies or antigen-binding fragments thereof of the present invention. The administration of therapeutic compositions in accordance with the invention will be administered with suitable carriers, excipients, and other agents that are incorporated into formulations to provide improved transfer, delivery, tolerance, and the like. A multitude of appropriate formulations can be found in the formulary known to all pharmaceutical chemists: Remington's Pharmaceutical Sciences, Mack Publishing Company, Easton, Pa. These formulations include, for example, powders, pastes, ointments, jellies, waxes, oils, lipids, lipid (cationic or anionic) containing vesicles (such as LIPOFECTIN®), DNA conjugates, anhydrous absorption pastes, oil-in-water and water-in-oil emulsions, emulsions carbowax (polyethylene glycols of various molecular weights), semi-solid gels, and semi-solid mixtures containing carbowax. See also Powell et al. "Compendium of excipients for parenteral formulations" PDA (1998) J Pharm Sci Technol 52:238-311.
[0054] The dose may vary depending upon the age and the size of a subject to be administered, target disease, conditions, route of administration, and the like. When the antibody of the present invention is used for treating various conditions and diseases associated with IL-4R, in an adult patient, it is advantageous to intravenously administer the antibody of the present invention normally at a single dose of about 0.01 to about 20 mg/kg body weight, more preferably about 0.02 to about 7, about 0.03 to about 5, or about 0.05 to about 3 mg/kg body weight. Depending on the severity of the condition, the frequency and the duration of the treatment can be adjusted.
[0055] Various delivery systems are known and can be used to administer the pharmaceutical composition of the invention, e.g., encapsulation in liposomes, microparticles, microcapsules, recombinant cells capable of expressing the mutant viruses, receptor mediated endocytosis (see, e.g., Wu et al. (1987) J. Biol. Chem. 262:4429-4432). Methods of introduction include, but are not limited to, intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal, epidural, and oral routes. The composition may be administered by any convenient route, for example by infusion or bolus injection, by absorption through epithelial or mucocutaneous linings (e.g., oral mucosa, rectal and intestinal mucosa, etc.) and may be administered together with other biologically active agents. Administration can be systemic or local.
[0056] The pharmaceutical composition can be also delivered in a vesicle, in particular a liposome (see Langer (1990) Science 249:1527-1533; Treat et al. (1989) in Liposomes in the Therapy of Infectious Disease and Cancer, Lopez Berestein and Fidler (eds.), Liss, New York, pp. 353-365; Lopez-Berestein, ibid., pp. 317-327; see generally ibid.
[0057] In certain situations, the pharmaceutical composition can be delivered in a controlled release system. In one embodiment, a pump may be used (see Langer, supra; Sefton (1987) CRC Crit. Ref. Biomed. Eng. 14:201). In another embodiment, polymeric materials can be used (see Medical Applications of Controlled Release, Langer and Wise (eds.), CRC Pres., Boca Raton, Fla. (1974). In yet another embodiment, a controlled release system can be placed in proximity of the composition's target, thus requiring only a fraction of the systemic dose (see, e.g., Goodson, in Medical Applications of Controlled Release, supra, vol. 2, pp. 115-138, 1984). Other controlled release systems are discussed in the review by Langer (1990) Science 249:1527-1533.
[0058] The injectable preparations may include dosage forms for intravenous, subcutaneous, intracutaneous and intramuscular injections, drip infusions, etc. These injectable preparations may be prepared by methods publicly known. For example, the injectable preparations may be prepared, e.g., by dissolving, suspending or emulsifying the antibody or its salt described above in a sterile aqueous medium or an oily medium conventionally used for injections. As the aqueous medium for injections, there are, for example, physiological saline, an isotonic solution containing glucose and other auxiliary agents, etc., which may be used in combination with an appropriate solubilizing agent such as an alcohol (e.g., ethanol), a polyalcohol (e.g., propylene glycol, polyethylene glycol), a nonionic surfactant [e.g., polysorbate 80, HCO-50 (polyoxyethylene (50 mol) adduct of hydrogenated castor oil)], etc. As the oily medium, there are employed, e.g., sesame oil, soybean oil, etc., which may be used in combination with a solubilizing agent such as benzyl benzoate, benzyl alcohol, etc. The injection thus prepared is preferably filled in an appropriate ampoule.
[0059] Advantageously, the pharmaceutical compositions for oral or parenteral use described above are prepared into dosage forms in a unit dose suited to fit a dose of the active ingredients. Such dosage forms in a unit dose include, for example, tablets, pills, capsules, injections (ampoules), suppositories, etc. The amount of the aforesaid antibody contained is generally about 5 to 500 mg per dosage form in a unit dose; especially in the form of injection, it is preferred that the aforesaid antibody is contained in about 5 to 100 mg and in about 10 to 250 mg for the other dosage forms.
[0060] Single and Combination Therapies.
[0061] The antibodies and antibody fragments of the invention are useful for treating diseases and disorders which are improved, inhibited or ameliorated by reducing IL-4 activity. These disorders include those characterized by abnormal or excess expression of IL-4, or by an abnormal host response to IL-4 production. IL-4 related disorders which are treated by the antibodies or antibody fragments of the include, for example, arthritis (including septic arthritis), herpetiformis, chronic idiopathic urticaria, scleroderma, hypertrophic scarring, Whipple's Disease, benign prostate hyperplasia, pulmonary disorders such as asthma (mild, moderate or severe), inflammatory disorders such as inflammatory bowel disease, allergic reactions, Kawasaki disease, sickle cell disease, Churg-Strauss syndrome, Grave's disease, pre-eclampsia, Sjogren's syndrome, autoimmune lymphoproliferative syndrome, autoimmune hemolytic anemia, Barrett's esophagus, autoimmune uveitis, tuberculosis, atopic dermatatis, ulcerative colitis, fibrosis, and nephrosis (see U.S. Pat. No. 7,186,809, herein specifically incorporated by reference).
[0062] The invention encompasses combination therapies in which the anti-IL-4R antibody or antibody fragment is administered in combination with a second therapeutic agent. Co-administration and combination therapy are not limited to simultaneous administration, but include treatment regimens in which an anti-IL-4R antibody or antibody fragment is administered at least once during a course of treatment that involves administering at least one other therapeutic agent to the patient. A second therapeutic agent may be another IL-4 antagonist, such as another antibody/antibody fragment, or a soluble cytokine receptor, an IgE antagonist, an anti-asthma medication (corticosteroids, non-steroidal agents, beta agonists, leukotriene antagonists, xanthines, fluticasone, salmeterol, albuterol) which may be delivered by inhalation or other appropriate means. In a specific embodiment, the anti-IL-4R antibody or antibody fragment of the invention may be administered with an IL-1 antagonist, such as rilonacept, or an IL-13 antagonist. The second agent may include one or more leukotriene receptor antagonists to treat disorders such as allergic inflammatory diseases, e.g., asthma and allergies. Examples of leukotriene receptor antagonists include but are not limited to montelukast, pranlukast, and zafirlukast. The second agent may include a cytokine inhibitor such as one or more of a TNF (etanercept, ENBREL®), IL-9, IL-5 or IL-17 antagonist.
EXAMPLES
[0063] The following examples are put forth so as to provide those of ordinary skill in the art with a complete disclosure and description of how to make and use the methods and compositions of the invention, and are not intended to limit the scope of what the inventors regard as their invention. Efforts have been made to ensure accuracy with respect to numbers used (e.g., amounts, temperature, etc.) but some experimental errors and deviations should be accounted for. Unless indicated otherwise, parts are parts by weight, molecular weight is average molecular weight, temperature is in degrees Centigrade, and pressure is at or near atmospheric.
Example 1
Generation of Human Antibodies to Human IL-4 Receptor
[0064] VELOCIMMUNE® mice (Regeneron Pharmaceuticals, Inc.; U.S. Pat. No. 6,596,541) were immunized with human IL-4R (hIL-4R, SEQ ID NO:274) or a combination of hIL-4R and monkey (Macaca fascicularis) IL-4R (mfIL-4R, SEQ ID NO:275) protein or DNA. To obtain optimal immune response, animals were subsequently boosted every 3-4 weeks and bleeds obtained 10 days after each boost for assessment of progression of anti-antigen response.
[0065] When the mice attained maximum immune response, antibody-expressing B cells were harvested and fused with mouse myeloma cells to form hybridomas. Alternatively, antigen-specific antibodies may be isolated directly from the B cells without fusion to myeloma cells, as described in U.S. Patent Publication 2007/0280945A1, herein specifically incorporated by reference in its entirety. Stable recombinant antibody-expressing CHO cell lines were established from the isolated proper recombinants. Functionally desirable monoclonal antibodies were selected by screening conditioned media of the hybridomas or transfected cells for specificity, antigen-binding affinity, and potency in blocking hIL-4 binding to hIL-4R (described below).
Example 2
Antigen Binding Affinity Determination
[0066] Binding affinity (KD) of selected antibodies with respect to hIL-4R at either 25° C. or 37° C. was determined using a real-time biosensor surface plasmon resonance assay (BIACORE® 2000). Briefly, antibody was captured on a goat anti-hFc polyclonal antibody surface created through direct coupling to a BIACORE® chip to form a captured antibody surface. Various concentrations (ranging from 50 nM to 12.5 nM) of monomeric hIL-4R(R&D Systems) or dimeric hIL-4R-mFc were injected over the captured antibody surface at 10 μl/min for 2.5 min at either 25° C. or 37° C. Binding of antigen to antibody and dissociation of the bound complex, were monitored in real time. Equilibrium dissociation constants (KD) and dissociation rate constants were ascertained by performing kinetic analysis using BIA evaluation software. BIA evaluation software was also used to calculate the half-life of antigen/antibody complex dissociation (T1/2). Results are shown in Table 1. NB: No antibody-antigen binding was observed under the experimental condition. Control: a fully human anti-IL-4R antibody (U.S. Pat. No. 7,186,809; SEQ ID NOs:10 and 12).
TABLE-US-00001 TABLE 1 25° C. 37° C. Monomeric Dimeric Monomeric Dimeric Antibody KD (pM) T1/2 (min) KD (pM) T1/2 (min) KD (pM) T1/2 (min) KD (pM) T1/2 (min) Control 1100 18 94 186 3970 4 114 158 H4H083P 48 361 28 245 183 87 38.1 163 H4H094P NB -- NB -- NB -- NB -- H4H095P 274 131 302 156 437 49 314 116 H4H098P 94.1 243 67.6 237 157 129 38.8 158 H4H099P NB -- NB -- NB -- NB --
[0067] Binding affinity (KD) of selected antibodies with respect to monkey (Macaca fascicularis) IL-4R (mfIL-4R) at either 25° C. or 37° C. was also determined using a real-time biosensor surface plasmon resonance assay described above with various concentrations (ranging from 100 nM to 25 nM) of monomeric mfIL-4R-myc-myc-his (mfIL-4R-mmh) or dimeric mfIL-4R-mFc. Only antibody H4H098P was able to bind both monomeric and dimeric mfIL-4R at 25° C. with KD of 552 nM and 9.08 nM, respectively. In addition, antibody H4H098P also binds to dimeric mfIL-4R at 37° C. with a KD of 24.3 nM. H4H083P had very weak binding to dimeric mfIL-4R.
[0068] Antibody-antigen binding affinity was also assessed using an ELISA-based solution competition assay. Briefly, a 96-well MAXISORP® plate was first coated with 5 g/ml avidin overnight followed by BSA blocking for 1 hr. The avidin-coated plate was then incubated with 250 ng/ml biotin-hlL4 for 2 hr. The plate was used to measure either free hIL-4R-mFc (dimeric hIL-4R) or free hIL-4R-myc-myc-his (hlL4R-mmh, monomeric hlL4R) in the antibody titration sample solutions. To make the antibody titration sample, a constant amount either 25 pM of hIL-4R-mFc or 200 pM of hIL-4R-mmh was premixed with varied amounts of antibody, ranging from 0 to about 10 nM in serial dilutions, followed by 1 hr incubation at room temperature to allow antibody-antigen-binding to reach equilibrium. The equilibrated sample solutions were then transferred to the hIL-4 coated plates for measurement of either free hIL-4R-mFc or free hIL-4R-mmh. After 1 hr binding, the plate was washed and bound hIL-4R-mFc was detected using either an HRP-conjugated mouse anti-mFc polyclonal antibody or an HRP-conjugated goat anti-myc polyclonal antibodies. IC50 values were determined (Table 2).
TABLE-US-00002 TABLE 2 IC50 (pM) Antibody 25 pM hIL-4R-mFc 200 pM hIL-4R-mmh Control 8.2 87 H4H083P 9.6 80 H4H094P >10,000 >10,000 H4H095P 40 90 H4H098P 8.8 74 H4H099P >10,000 >10,000
[0069] The ELISA-based solution competition assay was also used to determine the cross reactivity of the antibodies to monkey IL-4R. Antibody H4H098P exhibits an IC50 for mfIL-4R-mFc of 300 pM and an IC50 for mfIL-4R-mmh of 20 nM.
Example 3
Neutralization of Biological Effect of hIL-4 and hIL-13 In Vitro
[0070] A bioassay was developed to determine the purified anti-hIL-4R antibodies to neutralize hIL-4R-mediated cellular function in vitro using an engineered HK293 cell line that contains human STAT6 and a STAT6 luciferase reporter. Inhibition of hIL-4R-inducible luciferase activity was determined as follows: Cells were seeded onto 96-well plates at 1×104 cells/well in media and incubated overnight at 37° C., 5% CO2. Antibody proteins ranging from 0 to 20 nM in serial dilutions were added to the cells along with either 10 pM hIL-4 or 40 pM of hIL-13. Cells were then incubated at 37° C., 5% CO2 for 6 hrs. The extent of cellular response was measured in a luciferase assay (Promega Biotech). Results are shown in Table 3. NB: Luciferase activity was not blocked under the experimental condition described above. In addition, H4H098P was able to block mfIL-4R-mediated cellular function in the presence of 360 fM mfIL-4 with an IC50 of 150 nM.
TABLE-US-00003 TABLE 3 IC50 (pM) Antibody 10 pM hIL-4 40pM hIL-13 Control 47 38 H4H083P 25 19 H4H094P NB NB H4H095P 98 86 H4H098P 27 25 H4H099P NB 11,000
Sequence CWU
1
1
2751351DNAArtificial SequenceSynthetic 1caggtgcagc tggtggagtc tgggggaggc
gtggtccagc ctgggaggtc cctgagactc 60tcctgtgcag cctctggatt caccttccgc
tcttatggca tgcactgggt ccgccaggct 120ccaggcaagg ggctggagtg ggtggcggtc
atatcatatg atggaagtaa taaatattat 180atagactccg tgaagggccg attcaccatc
tccagagaca attccaagaa cacgctgaat 240ctgcaaatga acagcctgag acttgaggac
acggctgtat attactgtgc gaaagagggg 300agggggggat ttgactactg gggccaggga
atcccggtca ccgtctcctc a 3512117PRTArtificial
SequenceSynthetic 2Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln
Pro Gly Arg1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Arg Ser Tyr
20 25 30 Gly Met His Trp Val
Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40
45 Ala Val Ile Ser Tyr Asp Gly Ser Asn Lys
Tyr Tyr Ile Asp Ser Val 50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu
Asn65 70 75 80 Leu
Gln Met Asn Ser Leu Arg Leu Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95 Ala Lys Glu Gly Arg Gly
Gly Phe Asp Tyr Trp Gly Gln Gly Ile Pro 100
105 110 Val Thr Val Ser Ser 115
324DNAArtificial SequenceSynthetic 3ggattcacct tccgctctta tggc
2448PRTArtificial SequenceSynthetic 4Gly
Phe Thr Phe Arg Ser Tyr Gly1 5
524DNAArtificial SequenceSynthetic 5atatcatatg atggaagtaa taaa
2468PRTArtificial SequenceSynthetic 6Ile
Ser Tyr Asp Gly Ser Asn Lys1 5
730DNAArtificial SequenceSynthetic 7gcgaaagagg ggaggggggg atttgactac
30810PRTArtificial SequenceSynthetic 8Ala
Lys Glu Gly Arg Gly Gly Phe Asp Tyr1 5 10
9324DNAArtificial SequenceSynthetic 9gacatccaga tgacccagtc tccatcctca
ctgtctgcat ctgtaggaga cagagtcacc 60atcacttgtc gggcgagtca ggtcataaac
aattatttag cctggtttca gcagaaacca 120gggaaagtcc ctaagtccct gatccatgct
gcatccagtt tacaaagtgg ggtcccatca 180aagttcagcg gcagtggatc tgggacagat
ttcactctca ccatcagcag cctgcagcct 240gaagattttg caacttatta ctgccaacag
tataatagtc acccgtggac gttcggccaa 300gggaccaagg tggaaatcaa acga
32410108PRTArtificial SequenceSynthetic
10Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly1
5 10 15 Asp Arg Val Thr
Ile Thr Cys Arg Ala Ser Gln Val Ile Asn Asn Tyr 20
25 30 Leu Ala Trp Phe Gln Gln Lys Pro Gly
Lys Val Pro Lys Ser Leu Ile 35 40
45 His Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Lys Phe
Ser Gly 50 55 60
Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro65
70 75 80 Glu Asp Phe Ala Thr
Tyr Tyr Cys Gln Gln Tyr Asn Ser His Pro Trp 85
90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile
Lys Arg 100 105 1118DNAArtificial
SequenceSynthetic 11caggtcataa acaattat
18126PRTArtificial SequenceSynthetic 12Gln Val Ile Asn
Asn Tyr1 5 139DNAArtificial SequenceSynthetic
13gctgcatcc
9143PRTArtificial SequenceSynthetic 14Ala Ala Ser1
1527DNAArtificial SequenceSynthetic 15caacagtata atagtcaccc gtggacg
27169PRTArtificial SequenceSynthetic
16Gln Gln Tyr Asn Ser His Pro Trp Thr1 5
17351DNAArtificial SequenceSynthetic 17caggtgcagc tggtggagtc tgggggaggc
gtggtccagc ctgggaggtc cctgagactc 60tcctgtgcag cctctggatt caccttccgc
tcttatggca tgcactgggt ccgccaggct 120ccaggcaagg ggctggagtg ggtggcggtc
atatcatatg atggaagtaa taaatattat 180atagactccg tgaagggccg attcaccatc
tccagagaca attccaagaa cacgctgaat 240ctgcaaatga acagcctgag acttgaggac
acggctgtat attactgtgc gaaagagggg 300agggggggat ttgactactg gggccaggga
accctggtca ccgtctcctc a 35118117PRTArtificial
SequenceSynthetic 18Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln
Pro Gly Arg1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Arg Ser Tyr
20 25 30 Gly Met His Trp Val
Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40
45 Ala Val Ile Ser Tyr Asp Gly Ser Asn Lys
Tyr Tyr Ile Asp Ser Val 50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu
Asn65 70 75 80 Leu
Gln Met Asn Ser Leu Arg Leu Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95 Ala Lys Glu Gly Arg Gly
Gly Phe Asp Tyr Trp Gly Gln Gly Thr Leu 100
105 110 Val Thr Val Ser Ser 115
19321DNAArtificial SequenceSynthetic 19gacatccaga tgacccagtc tccatcctca
ctgtctgcat ctgtaggaga cagagtcacc 60atcacttgtc gggcgagtca ggtcataaac
aattatttag cctggtttca gcagaaacca 120gggaaagtcc ctaagtccct gatccatgct
gcatccagtt tacaaagtgg ggtcccatca 180aagttcagcg gcagtggatc tgggacagat
ttcactctca ccatcagcag cctgcagcct 240gaagattttg caacttatta ctgccaacag
tataatagtc acccgtggac gttcggccaa 300gggaccaagg tggaaatcaa a
32120107PRTArtificial SequenceSynthetic
20Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly1
5 10 15 Asp Arg Val Thr
Ile Thr Cys Arg Ala Ser Gln Val Ile Asn Asn Tyr 20
25 30 Leu Ala Trp Phe Gln Gln Lys Pro Gly
Lys Val Pro Lys Ser Leu Ile 35 40
45 His Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Lys Phe
Ser Gly 50 55 60
Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro65
70 75 80 Glu Asp Phe Ala Thr
Tyr Tyr Cys Gln Gln Tyr Asn Ser His Pro Trp 85
90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile
Lys 100 105 21351DNAArtificial
SequenceSynthetic 21caggtgcagc tggtggagtc tgggggaggc gtggtccagc
ctgggaggtc cctgagactc 60tcctgtgcag cctctggatt caccttccgc tcttatggca
tgcactgggt ccgccaggct 120ccaggcaagg ggctggagtg ggtggcagtt atatcatatg
atggaagtaa taaatactat 180gcagactccg tgaagggccg attcaccatc tccagagaca
attccaagaa cacgctgtat 240ctgcaaatga acagcctgag agctgaggac acggctgtgt
attactgtgc gaaagagggg 300agggggggat ttgactactg gggccaggga accctggtca
ccgtctcctc a 35122117PRTArtificial SequenceSynthetic 22Gln
Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg1
5 10 15 Ser Leu Arg Leu Ser Cys
Ala Ala Ser Gly Phe Thr Phe Arg Ser Tyr 20 25
30 Gly Met His Trp Val Arg Gln Ala Pro Gly Lys
Gly Leu Glu Trp Val 35 40 45
Ala Val Ile Ser Tyr Asp Gly Ser Asn Lys Tyr Tyr Ala Asp Ser Val
50 55 60 Lys Gly Arg
Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr65 70
75 80 Leu Gln Met Asn Ser Leu Arg Ala
Glu Asp Thr Ala Val Tyr Tyr Cys 85 90
95 Ala Lys Glu Gly Arg Gly Gly Phe Asp Tyr Trp Gly Gln
Gly Thr Leu 100 105 110
Val Thr Val Ser Ser 115 23322DNAArtificial
SequenceSynthetic 23gacatccaga tgacccagtc tccatcctca ctgtctgcat
ctgtaggaga cagagtcacc 60atcacttgtc gggcgagtca ggtcataaac aattatttag
cctggtttca gcagaaacca 120gggaaagccc ctaagtccct gatctatgct gcatccagtt
tgcaaagtgg ggtcccatca 180aggttcagcg gcagtggatc tgggacagat ttcactctca
ccatcagcag cctgcagcct 240gaagattttg caacttatta ctgccaacag tataatagtc
acccgtggac gttcggccaa 300gggaccaagg tggaaatcaa ac
32224107PRTArtificial SequenceSynthetic 24Asp Ile
Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly1 5
10 15 Asp Arg Val Thr Ile Thr Cys
Arg Ala Ser Gln Val Ile Asn Asn Tyr 20 25
30 Leu Ala Trp Phe Gln Gln Lys Pro Gly Lys Ala Pro
Lys Ser Leu Ile 35 40 45
Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly
50 55 60 Ser Gly Ser
Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro65 70
75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys
Gln Gln Tyr Asn Ser His Pro Trp 85 90
95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys
100 105 25351DNAArtificial SequenceSynthetic
25caggtgcagc tggtggagtc tgggggaggc gtggtccagc ctgggaggtc cctgagactc
60tcctgtgcag cctctggatt caccttcaga agctatggca tacactgggt ccgccaggct
120ccaggcaagg ggctggagtg ggtggcagtt atatcatatg atggaagtaa taaatactat
180gcagactccg tgaagggccg attcaccatc tccagagaca attccaagaa cacactgtat
240ctgcaaatga acagcctgat aactgaggac acggctgtgt attattgtgt gaaagagggg
300aggggggggt ttgactactg gggccaggga accacggtca ccgtctcctc a
35126117PRTArtificial SequenceSynthetic 26Gln Val Gln Leu Val Glu Ser Gly
Gly Gly Val Val Gln Pro Gly Arg1 5 10
15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe
Arg Ser Tyr 20 25 30
Gly Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45 Ala Val Ile Ser
Tyr Asp Gly Ser Asn Lys Tyr Tyr Ala Asp Ser Val 50 55
60 Lys Gly Arg Phe Thr Ile Ser Arg Asp
Asn Ser Lys Asn Thr Leu Tyr65 70 75
80 Leu Gln Met Asn Ser Leu Ile Thr Glu Asp Thr Ala Val Tyr
Tyr Cys 85 90 95
Val Lys Glu Gly Arg Gly Gly Phe Asp Tyr Trp Gly Gln Gly Thr Thr
100 105 110 Val Thr Val Ser Ser
115 2724DNAArtificial SequenceSynthetic 27ggattcacct
tcagaagcta tggc
24288PRTArtificial SequenceSynthetic 28Gly Phe Thr Phe Arg Ser Tyr Gly1
5 2924DNAArtificial SequenceSynthetic
29atatcatatg atggaagtaa taaa
24308PRTArtificial SequenceSynthetic 30Ile Ser Tyr Asp Gly Ser Asn Lys1
5 3130DNAArtificial SequenceSynthetic
31gtgaaagagg ggaggggggg gtttgactac
303210PRTArtificial SequenceSynthetic 32Val Lys Glu Gly Arg Gly Gly Phe
Asp Tyr1 5 10 33324DNAArtificial
SequenceSynthetic 33gacatccaga tgacccagtc tccatcctca ctgtctgcat
ctgtaggaga cagagtcacc 60atcacttgtc gggcgagtca ggtcattaat aattatttag
cctggtttca gcagaaacca 120gggaaagtcc ctaagtccct gatccatgct gcatccagtt
tgcaaagagg ggtcccatca 180aagttcagcg gcagtggatc tgggacagat ttcactctca
ccatcaacag cctgcagcct 240gaagattttg caacttatta ctgccaacaa tataatagtt
acccgtggac gttcggccaa 300gggaccaagg tggaaatcaa acga
32434108PRTArtificial SequenceSynthetic 34Asp Ile
Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly1 5
10 15 Asp Arg Val Thr Ile Thr Cys
Arg Ala Ser Gln Val Ile Asn Asn Tyr 20 25
30 Leu Ala Trp Phe Gln Gln Lys Pro Gly Lys Val Pro
Lys Ser Leu Ile 35 40 45
His Ala Ala Ser Ser Leu Gln Arg Gly Val Pro Ser Lys Phe Ser Gly
50 55 60 Ser Gly Ser
Gly Thr Asp Phe Thr Leu Thr Ile Asn Ser Leu Gln Pro65 70
75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys
Gln Gln Tyr Asn Ser Tyr Pro Trp 85 90
95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg
100 105 3518DNAArtificial
SequenceSynthetic 35caggtcatta ataattat
18366PRTArtificial SequenceSynthetic 36Gln Val Ile Asn
Asn Tyr1 5 379DNAArtificial SequenceSynthetic
37gctgcatcc
9383PRTArtificial SequenceSynthetic 38Ala Ala Ser1
3927DNAArtificial SequenceSynthetic 39caacaatata atagttaccc gtggacg
27409PRTArtificial SequenceSynthetic
40Gln Gln Tyr Asn Ser Tyr Pro Trp Thr1 5
41351DNAArtificial SequenceSynthetic 41caggtgcagc tggtggagtc tgggggaggc
gtggtccagc ctgggaggtc cctgagactc 60tcctgtgcag cctctggatt caccttcaga
agctatggca tacactgggt ccgccaggct 120ccaggcaagg ggctggagtg ggtggcagtt
atatcatatg atggaagtaa taaatactat 180gcagactccg tgaagggccg attcaccatc
tccagagaca attccaagaa cacactgtat 240ctgcaaatga acagcctgat aactgaggac
acggctgtgt attattgtgt gaaagagggg 300aggggggggt ttgactactg gggccaggga
accctggtca ccgtctcctc a 35142117PRTArtificial
SequenceSynthetic 42Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln
Pro Gly Arg1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Arg Ser Tyr
20 25 30 Gly Ile His Trp Val
Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40
45 Ala Val Ile Ser Tyr Asp Gly Ser Asn Lys
Tyr Tyr Ala Asp Ser Val 50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu
Tyr65 70 75 80 Leu
Gln Met Asn Ser Leu Ile Thr Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95 Val Lys Glu Gly Arg Gly
Gly Phe Asp Tyr Trp Gly Gln Gly Thr Leu 100
105 110 Val Thr Val Ser Ser 115
43321DNAArtificial SequenceSynthetic 43gacatccaga tgacccagtc tccatcctca
ctgtctgcat ctgtaggaga cagagtcacc 60atcacttgtc gggcgagtca ggtcattaat
aattatttag cctggtttca gcagaaacca 120gggaaagtcc ctaagtccct gatccatgct
gcatccagtt tgcaaagagg ggtcccatca 180aagttcagcg gcagtggatc tgggacagat
ttcactctca ccatcaacag cctgcagcct 240gaagattttg caacttatta ctgccaacaa
tataatagtt acccgtggac gttcggccaa 300gggaccaagg tggaaatcaa a
32144107PRTArtificial SequenceSynthetic
44Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly1
5 10 15 Asp Arg Val Thr
Ile Thr Cys Arg Ala Ser Gln Val Ile Asn Asn Tyr 20
25 30 Leu Ala Trp Phe Gln Gln Lys Pro Gly
Lys Val Pro Lys Ser Leu Ile 35 40
45 His Ala Ala Ser Ser Leu Gln Arg Gly Val Pro Ser Lys Phe
Ser Gly 50 55 60
Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Asn Ser Leu Gln Pro65
70 75 80 Glu Asp Phe Ala Thr
Tyr Tyr Cys Gln Gln Tyr Asn Ser Tyr Pro Trp 85
90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile
Lys 100 105 45351DNAArtificial
SequenceSynthetic 45caggtgcagc tggtggagtc tgggggaggc gtggtccagc
ctgggaggtc cctgagactc 60tcctgtgcag cctctggatt caccttcaga agctatggca
tgcactgggt ccgccaggct 120ccaggcaagg ggctggagtg ggtggcagtt atatcatatg
atggaagtaa taaatactat 180gcagactccg tgaagggccg attcaccatc tccagagaca
attccaagaa cacgctgtat 240ctgcaaatga acagcctgag agctgaggac acggctgtgt
attactgtgt gaaagagggg 300aggggggggt ttgactactg gggccaggga accctggtca
ccgtctcctc a 35146117PRTArtificial SequenceSynthetic 46Gln
Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg1
5 10 15 Ser Leu Arg Leu Ser Cys
Ala Ala Ser Gly Phe Thr Phe Arg Ser Tyr 20 25
30 Gly Met His Trp Val Arg Gln Ala Pro Gly Lys
Gly Leu Glu Trp Val 35 40 45
Ala Val Ile Ser Tyr Asp Gly Ser Asn Lys Tyr Tyr Ala Asp Ser Val
50 55 60 Lys Gly Arg
Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr65 70
75 80 Leu Gln Met Asn Ser Leu Arg Ala
Glu Asp Thr Ala Val Tyr Tyr Cys 85 90
95 Val Lys Glu Gly Arg Gly Gly Phe Asp Tyr Trp Gly Gln
Gly Thr Leu 100 105 110
Val Thr Val Ser Ser 115 47322DNAArtificial
SequenceSynthetic 47gacatccaga tgacccagtc tccatcctca ctgtctgcat
ctgtaggaga cagagtcacc 60atcacttgtc gggcgagtca ggtcattaat aattatttag
cctggtttca gcagaaacca 120gggaaagccc ctaagtccct gatctatgct gcatccagtt
tgcaaagtgg ggtcccatca 180aggttcagcg gcagtggatc tgggacagat ttcactctca
ccatcagcag cctgcagcct 240gaagattttg caacttatta ctgccaacaa tataatagtt
acccgtggac gttcggccaa 300gggaccaagg tggaaatcaa ac
32248107PRTArtificial SequenceSynthetic 48Asp Ile
Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly1 5
10 15 Asp Arg Val Thr Ile Thr Cys
Arg Ala Ser Gln Val Ile Asn Asn Tyr 20 25
30 Leu Ala Trp Phe Gln Gln Lys Pro Gly Lys Ala Pro
Lys Ser Leu Ile 35 40 45
Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly
50 55 60 Ser Gly Ser
Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro65 70
75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys
Gln Gln Tyr Asn Ser Tyr Pro Trp 85 90
95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys
100 105 49375DNAArtificial SequenceSynthetic
49caggtgcagc tggtggagtc tgggggaggc ttggaacagc cgggggggtc cttgagactc
60tcctgtgcag gctctggatt cacgtttaga gactatgcca tgacctgggt ccgccaggct
120ccagggaagg ggctggagtg ggtcgcatcg attagtggtt ccggtggtaa cacatacttc
180gcagactccg tgaagggccg gttcaccatc tccagagaca attccaagaa cacgctgtat
240ctgcaaatga acagcctgag agccgaggac acggccgtat attactgtgc gaaagatcga
300ctctctataa caattcgccc acgctattat ggtttggacg tctggggcca agggtccacg
360gtcaccgtct cctca
37550125PRTArtificial SequenceSynthetic 50Gln Val Gln Leu Val Glu Ser Gly
Gly Gly Leu Glu Gln Pro Gly Gly1 5 10
15 Ser Leu Arg Leu Ser Cys Ala Gly Ser Gly Phe Thr Phe
Arg Asp Tyr 20 25 30
Ala Met Thr Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45 Ala Ser Ile Ser
Gly Ser Gly Gly Asn Thr Tyr Phe Ala Asp Ser Val 50 55
60 Lys Gly Arg Phe Thr Ile Ser Arg Asp
Asn Ser Lys Asn Thr Leu Tyr65 70 75
80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr
Tyr Cys 85 90 95
Ala Lys Asp Arg Leu Ser Ile Thr Ile Arg Pro Arg Tyr Tyr Gly Leu
100 105 110 Asp Val Trp Gly Gln
Gly Ser Thr Val Thr Val Ser Ser 115 120
125 5124DNAArtificial SequenceSynthetic 51ggattcacgt ttagagacta tgcc
24528PRTArtificial
SequenceSynthetic 52Gly Phe Thr Phe Arg Asp Tyr Ala1 5
5324DNAArtificial SequenceSynthetic 53attagtggtt ccggtggtaa caca
24548PRTArtificial
SequenceSynthetic 54Ile Ser Gly Ser Gly Gly Asn Thr1 5
5554DNAArtificial SequenceSynthetic 55gcgaaagatc gactctctat
aacaattcgc ccacgctatt atggtttgga cgtc 545618PRTArtificial
SequenceSynthetic 56Ala Lys Asp Arg Leu Ser Ile Thr Ile Arg Pro Arg Tyr
Tyr Gly Leu1 5 10 15
Asp Val57324DNAArtificial SequenceSynthetic 57gacatccaga tgacccagtc
tccatcctca ctgtctgcat ctgttggaga cagagtcacc 60atcacttgtc gggcgagtca
ggccattaac aatcatttag cctggtttca gcagaaacca 120gggaaagccc ctaagtccct
gatctttgct gtatccagtt tgcaaagtgg ggtcccatca 180aagttcagcg gcagtggatc
tgggacagac ttcactctca ccatcagcag cctgcagcct 240gaagattttg caacttatta
ctgccaacag tataatagtt acccgtggac gttcggccaa 300gggaccaagg tggaaatcaa
acga 32458108PRTArtificial
SequenceSynthetic 58Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala
Ser Val Gly1 5 10 15
Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ala Ile Asn Asn His
20 25 30 Leu Ala Trp Phe Gln
Gln Lys Pro Gly Lys Ala Pro Lys Ser Leu Ile 35 40
45 Phe Ala Val Ser Ser Leu Gln Ser Gly Val
Pro Ser Lys Phe Ser Gly 50 55 60
Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln
Pro65 70 75 80 Glu
Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Asn Ser Tyr Pro Trp
85 90 95 Thr Phe Gly Gln Gly Thr
Lys Val Glu Ile Lys Arg 100 105
5918DNAArtificial SequenceSynthetic 59caggccatta acaatcat
18606PRTArtificial SequenceSynthetic
60Gln Ala Ile Asn Asn His1 5 619DNAArtificial
SequenceSynthetic 61gctgtatcc
9623PRTArtificial SequenceSynthetic 62Ala Val Ser1
6327DNAArtificial SequenceSynthetic 63caacagtata atagttaccc gtggacg
27649PRTArtificial SequenceSynthetic
64Gln Gln Tyr Asn Ser Tyr Pro Trp Thr1 5
65372DNAArtificial SequenceSynthetic 65gaggtgcagc tggtggagtc tgggggaggc
ttggaacagc cgggggggtc cttgagactc 60tcctgtgcag gctctggatt cacgtttaga
gactatgcca tgacctgggt ccgccaggct 120ccagggaagg ggctggagtg ggtcgcatcg
attagtggtt ccggtggtaa cacatacttc 180gcagactccg tgaagggccg gttcaccatc
tccagagaca attccaagaa cacgctgtat 240ctgcaaatga acagcctgag agccgaggac
acggccgtat attactgtgc gaaagatcga 300ctctctataa caattcgccc acgctattat
ggtttggacg tctggggcca agggaccacg 360gtcaccgtct cc
37266124PRTArtificial SequenceSynthetic
66Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Glu Gln Pro Gly Gly1
5 10 15 Ser Leu Arg Leu
Ser Cys Ala Gly Ser Gly Phe Thr Phe Arg Asp Tyr 20
25 30 Ala Met Thr Trp Val Arg Gln Ala Pro
Gly Lys Gly Leu Glu Trp Val 35 40
45 Ala Ser Ile Ser Gly Ser Gly Gly Asn Thr Tyr Phe Ala Asp
Ser Val 50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr65
70 75 80 Leu Gln Met Asn Ser
Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85
90 95 Ala Lys Asp Arg Leu Ser Ile Thr Ile Arg
Pro Arg Tyr Tyr Gly Leu 100 105
110 Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser 115
120 67321DNAArtificial SequenceSynthetic
67gacatccaga tgacccagtc tccatcctca ctgtctgcat ctgttggaga cagagtcacc
60atcacttgtc gggcgagtca ggccattaac aatcatttag cctggtttca gcagaaacca
120gggaaagccc ctaagtccct gatctttgct gtatccagtt tgcaaagtgg ggtcccatca
180aagttcagcg gcagtggatc tgggacagac ttcactctca ccatcagcag cctgcagcct
240gaagattttg caacttatta ctgccaacag tataatagtt acccgtggac gttcggccaa
300gggaccaagg tggaaatcaa a
32168107PRTArtificial SequenceSynthetic 68Asp Ile Gln Met Thr Gln Ser Pro
Ser Ser Leu Ser Ala Ser Val Gly1 5 10
15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ala Ile
Asn Asn His 20 25 30
Leu Ala Trp Phe Gln Gln Lys Pro Gly Lys Ala Pro Lys Ser Leu Ile
35 40 45 Phe Ala Val Ser
Ser Leu Gln Ser Gly Val Pro Ser Lys Phe Ser Gly 50 55
60 Ser Gly Ser Gly Thr Asp Phe Thr Leu
Thr Ile Ser Ser Leu Gln Pro65 70 75
80 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Asn Ser Tyr
Pro Trp 85 90 95
Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100
105 69373DNAArtificial SequenceSynthetic 69gaggtgcagc tggtggagtc
tgggggaggc ttggtacagc ctggggggtc cctgagactc 60tcctgtgcag cctctggatt
cacgtttaga gactatgcca tgagctgggt ccgccaggct 120ccagggaagg ggctggagtg
ggtctcagct attagtggtt ccggtggtaa cacatactac 180gcagactccg tgaagggccg
gttcaccatc tccagagaca attccaagaa cacgctgtat 240ctgcaaatga acagcctgag
agccgaggac acggccgtat attactgtgc gaaagatcga 300ctctctataa caattcgccc
acgctattat ggtttggacg tctggggcca agggaccacg 360gtcaccgtct cct
37370124PRTArtificial
SequenceSynthetic 70Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln
Pro Gly Gly1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Arg Asp Tyr
20 25 30 Ala Met Ser Trp Val
Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40
45 Ser Ala Ile Ser Gly Ser Gly Gly Asn Thr
Tyr Tyr Ala Asp Ser Val 50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu
Tyr65 70 75 80 Leu
Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95 Ala Lys Asp Arg Leu Ser
Ile Thr Ile Arg Pro Arg Tyr Tyr Gly Leu 100
105 110 Asp Val Trp Gly Gln Gly Thr Thr Val Thr
Val Ser 115 120 71322DNAArtificial
SequenceSynthetic 71gacatccaga tgacccagtc tccatcctca ctgtctgcat
ctgtaggaga cagagtcacc 60atcacttgtc gggcgagtca ggccattaac aatcatttag
cctggtttca gcagaaacca 120gggaaagccc ctaagtccct gatctatgct gtatccagtt
tgcaaagtgg ggtcccatca 180aggttcagcg gcagtggatc tgggacagat ttcactctca
ccatcagcag cctgcagcct 240gaagattttg caacttatta ctgccaacag tataatagtt
acccgtggac gttcggccaa 300gggaccaagg tggaaatcaa ac
32272107PRTArtificial SequenceSynthetic 72Asp Ile
Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly1 5
10 15 Asp Arg Val Thr Ile Thr Cys
Arg Ala Ser Gln Ala Ile Asn Asn His 20 25
30 Leu Ala Trp Phe Gln Gln Lys Pro Gly Lys Ala Pro
Lys Ser Leu Ile 35 40 45
Tyr Ala Val Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly
50 55 60 Ser Gly Ser
Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro65 70
75 80 Glu Asp Phe Ala Thr Tyr Tyr Cys
Gln Gln Tyr Asn Ser Tyr Pro Trp 85 90
95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys
100 105 73375DNAArtificial SequenceSynthetic
73caggtgcagc tggtggagtc tgggggaggc ttggaacagc cgggggggtc cttgagactc
60tcctgtgcag gctctggatt cacgtttaga gactatgcca tgacctgggt ccgccaggct
120ccagggaagg ggctggagtg ggtcgcatcg attagtggtt ccggtggtaa cacatacttc
180gcagactccg tgaagggccg gttcaccatc tccagagaca attccaagaa cacgctgtat
240ctgcaaatga acagcctgag agccgaggac acggccgtat attactgtgc gaaagatcga
300ctctctataa caattcgccc acgctattat ggtttggacg tctggggcca agggtccacg
360gtcaccgtct cctca
37574125PRTArtificial SequenceSynthetic 74Gln Val Gln Leu Val Glu Ser Gly
Gly Gly Leu Glu Gln Pro Gly Gly1 5 10
15 Ser Leu Arg Leu Ser Cys Ala Gly Ser Gly Phe Thr Phe
Arg Asp Tyr 20 25 30
Ala Met Thr Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45 Ala Ser Ile Ser
Gly Ser Gly Gly Asn Thr Tyr Phe Ala Asp Ser Val 50 55
60 Lys Gly Arg Phe Thr Ile Ser Arg Asp
Asn Ser Lys Asn Thr Leu Tyr65 70 75
80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr
Tyr Cys 85 90 95
Ala Lys Asp Arg Leu Ser Ile Thr Ile Arg Pro Arg Tyr Tyr Gly Leu
100 105 110 Asp Val Trp Gly Gln
Gly Ser Thr Val Thr Val Ser Ser 115 120
125 7524DNAArtificial SequenceSynthetic 75ggattcacgt ttagagacta tgcc
24768PRTArtificial
SequenceSynthetic 76Gly Phe Thr Phe Arg Asp Tyr Ala1 5
7724DNAArtificial SequenceSynthetic 77attagtggtt ccggtggtaa caca
24788PRTArtificial
SequenceSynthetic 78Ile Ser Gly Ser Gly Gly Asn Thr1 5
7954DNAArtificial SequenceSynthetic 79gcgaaagatc gactctctat
aacaattcgc ccacgctatt atggtttgga cgtc 548018PRTArtificial
SequenceSynthetic 80Ala Lys Asp Arg Leu Ser Ile Thr Ile Arg Pro Arg Tyr
Tyr Gly Leu1 5 10 15
Asp Val81339DNAArtificial SequenceSynthetic 81gaaatagtgt tgacgcagtc
tccactctcc ctgcccgtca cccctggaga gccggcctcc 60atctcctgca ggtctagtca
gagcctcctg tatagtattg gatacaacta tttggattgg 120tacctgcaga agtcagggca
gtctccacag ctccttatct atttgggttc taatcgggcc 180tccggggtcc ctgacaggtt
cagtggcagt ggatcaggca cagattttac actgaaaatc 240agcagagtgg aggctgagga
tgttgggttt tattactgca tgcaagctct acaaactccg 300tacacttttg gcccggggac
caagctggag atcaaacga 33982113PRTArtificial
SequenceSynthetic 82Glu Ile Val Leu Thr Gln Ser Pro Leu Ser Leu Pro Val
Thr Pro Gly1 5 10 15
Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Leu Tyr Ser
20 25 30 Ile Gly Tyr Asn Tyr
Leu Asp Trp Tyr Leu Gln Lys Ser Gly Gln Ser 35 40
45 Pro Gln Leu Leu Ile Tyr Leu Gly Ser Asn
Arg Ala Ser Gly Val Pro 50 55 60
Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys
Ile65 70 75 80 Ser
Arg Val Glu Ala Glu Asp Val Gly Phe Tyr Tyr Cys Met Gln Ala
85 90 95 Leu Gln Thr Pro Tyr Thr
Phe Gly Pro Gly Thr Lys Leu Glu Ile Lys 100
105 110 Arg8333DNAArtificial SequenceSynthetic
83cagagcctcc tgtatagtat tggatacaac tat
338411PRTArtificial SequenceSynthetic 84Gln Ser Leu Leu Tyr Ser Ile Gly
Tyr Asn Tyr1 5 10 859DNAArtificial
SequenceSynthetic 85ttgggttct
9863PRTArtificial SequenceSynthetic 86Leu Gly Ser1
8727DNAArtificial SequenceSynthetic 87atgcaagctc tacaaactcc gtacact
27889PRTArtificial SequenceSynthetic
88Met Gln Ala Leu Gln Thr Pro Tyr Thr1 5
89372DNAArtificial SequenceSynthetic 89gaggtgcagc tggtggagtc tgggggaggc
ttggaacagc cgggggggtc cttgagactc 60tcctgtgcag gctctggatt cacgtttaga
gactatgcca tgacctgggt ccgccaggct 120ccagggaagg ggctggagtg ggtcgcatcg
attagtggtt ccggtggtaa cacatacttc 180gcagactccg tgaagggccg gttcaccatc
tccagagaca attccaagaa cacgctgtat 240ctgcaaatga acagcctgag agccgaggac
acggccgtat attactgtgc gaaagatcga 300ctctctataa caattcgccc acgctattat
ggtttggacg tctggggcca agggaccacg 360gtcaccgtct cc
37290124PRTArtificial SequenceSynthetic
90Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Glu Gln Pro Gly Gly1
5 10 15 Ser Leu Arg Leu
Ser Cys Ala Gly Ser Gly Phe Thr Phe Arg Asp Tyr 20
25 30 Ala Met Thr Trp Val Arg Gln Ala Pro
Gly Lys Gly Leu Glu Trp Val 35 40
45 Ala Ser Ile Ser Gly Ser Gly Gly Asn Thr Tyr Phe Ala Asp
Ser Val 50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr65
70 75 80 Leu Gln Met Asn Ser
Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85
90 95 Ala Lys Asp Arg Leu Ser Ile Thr Ile Arg
Pro Arg Tyr Tyr Gly Leu 100 105
110 Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser 115
120 91336DNAArtificial SequenceSynthetic
91gatattgtga tgactcagtc tccactctcc ctgcccgtca cccctggaga gccggcctcc
60atctcctgca ggtctagtca gagcctcctg tatagtattg gatacaacta tttggattgg
120tacctgcaga agtcagggca gtctccacag ctccttatct atttgggttc taatcgggcc
180tccggggtcc ctgacaggtt cagtggcagt ggatcaggca cagattttac actgaaaatc
240agcagagtgg aggctgagga tgttgggttt tattactgca tgcaagctct acaaactccg
300tacacttttg gcccggggac caagctggag atcaaa
33692112PRTArtificial SequenceSynthetic 92Asp Ile Val Met Thr Gln Ser Pro
Leu Ser Leu Pro Val Thr Pro Gly1 5 10
15 Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu
Leu Tyr Ser 20 25 30
Ile Gly Tyr Asn Tyr Leu Asp Trp Tyr Leu Gln Lys Ser Gly Gln Ser
35 40 45 Pro Gln Leu Leu
Ile Tyr Leu Gly Ser Asn Arg Ala Ser Gly Val Pro 50 55
60 Asp Arg Phe Ser Gly Ser Gly Ser Gly
Thr Asp Phe Thr Leu Lys Ile65 70 75
80 Ser Arg Val Glu Ala Glu Asp Val Gly Phe Tyr Tyr Cys Met
Gln Ala 85 90 95
Leu Gln Thr Pro Tyr Thr Phe Gly Pro Gly Thr Lys Leu Glu Ile Lys
100 105 110 93373DNAArtificial
SequenceSynthetic 93gaggtgcagc tggtggagtc tgggggaggc ttggtacagc
ctggggggtc cctgagactc 60tcctgtgcag cctctggatt cacgtttaga gactatgcca
tgagctgggt ccgccaggct 120ccagggaagg ggctggagtg ggtctcagct attagtggtt
ccggtggtaa cacatactac 180gcagactccg tgaagggccg gttcaccatc tccagagaca
attccaagaa cacgctgtat 240ctgcaaatga acagcctgag agccgaggac acggccgtat
attactgtgc gaaagatcga 300ctctctataa caattcgccc acgctattat ggtttggacg
tctggggcca agggaccacg 360gtcaccgtct cct
37394124PRTArtificial SequenceSynthetic 94Glu Val
Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly1 5
10 15 Ser Leu Arg Leu Ser Cys Ala
Ala Ser Gly Phe Thr Phe Arg Asp Tyr 20 25
30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly
Leu Glu Trp Val 35 40 45
Ser Ala Ile Ser Gly Ser Gly Gly Asn Thr Tyr Tyr Ala Asp Ser Val
50 55 60 Lys Gly Arg
Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr65 70
75 80 Leu Gln Met Asn Ser Leu Arg Ala
Glu Asp Thr Ala Val Tyr Tyr Cys 85 90
95 Ala Lys Asp Arg Leu Ser Ile Thr Ile Arg Pro Arg Tyr
Tyr Gly Leu 100 105 110
Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser 115
120 95337DNAArtificial SequenceSynthetic 95gatattgtga
tgactcagtc tccactctcc ctgcccgtca cccctggaga gccggcctcc 60atctcctgca
ggtctagtca gagcctcctg tatagtattg gatacaacta tttggattgg 120tacctgcaga
agccagggca gtctccacag ctcctgatct atttgggttc taatcgggcc 180tccggggtcc
ctgacaggtt cagtggcagt ggatcaggca cagattttac actgaaaatc 240agcagagtgg
aggctgagga tgttggggtt tattactgca tgcaagctct acaaactccg 300tacacttttg
gccaggggac caagctggag atcaaac
33796112PRTArtificial SequenceSynthetic 96Asp Ile Val Met Thr Gln Ser Pro
Leu Ser Leu Pro Val Thr Pro Gly1 5 10
15 Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu
Leu Tyr Ser 20 25 30
Ile Gly Tyr Asn Tyr Leu Asp Trp Tyr Leu Gln Lys Pro Gly Gln Ser
35 40 45 Pro Gln Leu Leu
Ile Tyr Leu Gly Ser Asn Arg Ala Ser Gly Val Pro 50 55
60 Asp Arg Phe Ser Gly Ser Gly Ser Gly
Thr Asp Phe Thr Leu Lys Ile65 70 75
80 Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Met
Gln Ala 85 90 95
Leu Gln Thr Pro Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys
100 105 110 97375DNAArtificial
SequenceSynthetic 97caggtgcagc tggtggagtc tgagggactc ttggaacagc
ctggggggtc cctgagactc 60tcctgtgcag cctctggatt caactttaga gactttgcca
tgacctgggt ccgccaggct 120ccagggaagg ggctggagtg ggtctcatct attagtggta
gtggtagtaa tacatactac 180gcagactccg tgaagggccg gttcaccatc tccagagaca
attccaacca cacgctgtat 240ctgcgaatga acagcctgag agccgaagac acggccgtgt
attactgtgc gaaagatcga 300ctctctataa caattcgccc acgctattac ggtctggacg
tctggggcca agggtccacg 360gtcaccgtct cctca
37598125PRTArtificial SequenceSynthetic 98Gln Val
Gln Leu Val Glu Ser Glu Gly Leu Leu Glu Gln Pro Gly Gly1 5
10 15 Ser Leu Arg Leu Ser Cys Ala
Ala Ser Gly Phe Asn Phe Arg Asp Phe 20 25
30 Ala Met Thr Trp Val Arg Gln Ala Pro Gly Lys Gly
Leu Glu Trp Val 35 40 45
Ser Ser Ile Ser Gly Ser Gly Ser Asn Thr Tyr Tyr Ala Asp Ser Val
50 55 60 Lys Gly Arg
Phe Thr Ile Ser Arg Asp Asn Ser Asn His Thr Leu Tyr65 70
75 80 Leu Arg Met Asn Ser Leu Arg Ala
Glu Asp Thr Ala Val Tyr Tyr Cys 85 90
95 Ala Lys Asp Arg Leu Ser Ile Thr Ile Arg Pro Arg Tyr
Tyr Gly Leu 100 105 110
Asp Val Trp Gly Gln Gly Ser Thr Val Thr Val Ser Ser 115
120 125 9924DNAArtificial SequenceSynthetic
99ggattcaact ttagagactt tgcc
241008PRTArtificial SequenceSynthetic 100Gly Phe Asn Phe Arg Asp Phe Ala1
5 10124DNAArtificial SequenceSynthetic
101attagtggta gtggtagtaa taca
241028PRTArtificial SequenceSynthetic 102Ile Ser Gly Ser Gly Ser Asn Thr1
5 10354DNAArtificial SequenceSynthetic
103gcgaaagatc gactctctat aacaattcgc ccacgctatt acggtctgga cgtc
5410418PRTArtificial SequenceSynthetic 104Ala Lys Asp Arg Leu Ser Ile Thr
Ile Arg Pro Arg Tyr Tyr Gly Leu1 5 10
15 Asp Val105324DNAArtificial SequenceSynthetic
105gacatccaga tgacccagtc tccatcctcc ctgtctgcat ctgtaggaga cagagtcacc
60atcacttgcc gggcgagtca ggacattagc aattattttg cctggtatca gcagaagcca
120gggaaagttc ctaagctcct gatctttgct gcatccactt tgcatccagg ggtcccatct
180cggttcagtg gcagtggatc tgggacagat ttcactctca ccattcgcag cctgcagcct
240gaagatgttg caacttatta ctgtcaaaaa tatgacagtg ccccgtacac ttttggccag
300gggaccaagg tggaaatcaa acga
324106108PRTArtificial SequenceSynthetic 106Asp Ile Gln Met Thr Gln Ser
Pro Ser Ser Leu Ser Ala Ser Val Gly1 5 10
15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp
Ile Ser Asn Tyr 20 25 30
Phe Ala Trp Tyr Gln Gln Lys Pro Gly Lys Val Pro Lys Leu Leu Ile
35 40 45 Phe Ala Ala Ser
Thr Leu His Pro Gly Val Pro Ser Arg Phe Ser Gly 50 55
60 Ser Gly Ser Gly Thr Asp Phe Thr Leu
Thr Ile Arg Ser Leu Gln Pro65 70 75
80 Glu Asp Val Ala Thr Tyr Tyr Cys Gln Lys Tyr Asp Ser Ala
Pro Tyr 85 90 95
Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg 100
105 10718DNAArtificial SequenceSynthetic 107caggacatta
gcaattat
181086PRTArtificial SequenceSynthetic 108Gln Asp Ile Ser Asn Tyr1
5 1099DNAArtificial SequenceSynthetic 109gctgcatcc
91103PRTArtificial
SequenceSynthetic 110Ala Ala Ser1 11127DNAArtificial
SequenceSynthetic 111caaaaatatg acagtgcccc gtacact
271129PRTArtificial SequenceSynthetic 112Gln Lys Tyr Asp
Ser Ala Pro Tyr Thr1 5 113372DNAArtificial
SequenceSynthetic 113gaggtgcagc tggtggagtc tgagggactc ttggaacagc
ctggggggtc cctgagactc 60tcctgtgcag cctctggatt caactttaga gactttgcca
tgacctgggt ccgccaggct 120ccagggaagg ggctggagtg ggtctcatct attagtggta
gtggtagtaa tacatactac 180gcagactccg tgaagggccg gttcaccatc tccagagaca
attccaacca cacgctgtat 240ctgcgaatga acagcctgag agccgaagac acggccgtgt
attactgtgc gaaagatcga 300ctctctataa caattcgccc acgctattac ggtctggacg
tctggggcca agggaccacg 360gtcaccgtct cc
372114124PRTArtificial SequenceSynthetic 114Glu
Val Gln Leu Val Glu Ser Glu Gly Leu Leu Glu Gln Pro Gly Gly1
5 10 15 Ser Leu Arg Leu Ser Cys
Ala Ala Ser Gly Phe Asn Phe Arg Asp Phe 20 25
30 Ala Met Thr Trp Val Arg Gln Ala Pro Gly Lys
Gly Leu Glu Trp Val 35 40 45
Ser Ser Ile Ser Gly Ser Gly Ser Asn Thr Tyr Tyr Ala Asp Ser Val
50 55 60 Lys Gly Arg
Phe Thr Ile Ser Arg Asp Asn Ser Asn His Thr Leu Tyr65 70
75 80 Leu Arg Met Asn Ser Leu Arg Ala
Glu Asp Thr Ala Val Tyr Tyr Cys 85 90
95 Ala Lys Asp Arg Leu Ser Ile Thr Ile Arg Pro Arg Tyr
Tyr Gly Leu 100 105 110
Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser 115
120 115321DNAArtificial SequenceSynthetic 115gacatccaga
tgacccagtc tccatcctcc ctgtctgcat ctgtaggaga cagagtcacc 60atcacttgcc
gggcgagtca ggacattagc aattattttg cctggtatca gcagaagcca 120gggaaagttc
ctaagctcct gatctttgct gcatccactt tgcatccagg ggtcccatct 180cggttcagtg
gcagtggatc tgggacagat ttcactctca ccattcgcag cctgcagcct 240gaagatgttg
caacttatta ctgtcaaaaa tatgacagtg ccccgtacac ttttggccag 300gggaccaagc
tggagatcaa a
321116107PRTArtificial SequenceSynthetic 116Asp Ile Gln Met Thr Gln Ser
Pro Ser Ser Leu Ser Ala Ser Val Gly1 5 10
15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp
Ile Ser Asn Tyr 20 25 30
Phe Ala Trp Tyr Gln Gln Lys Pro Gly Lys Val Pro Lys Leu Leu Ile
35 40 45 Phe Ala Ala Ser
Thr Leu His Pro Gly Val Pro Ser Arg Phe Ser Gly 50 55
60 Ser Gly Ser Gly Thr Asp Phe Thr Leu
Thr Ile Arg Ser Leu Gln Pro65 70 75
80 Glu Asp Val Ala Thr Tyr Tyr Cys Gln Lys Tyr Asp Ser Ala
Pro Tyr 85 90 95
Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100
105 117373DNAArtificial SequenceSynthetic 117gaggtgcagc
tggtggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60tcctgtgcag
cctctggatt caactttaga gactttgcca tgagctgggt ccgccaggct 120ccagggaagg
ggctggagtg ggtctcagct attagtggta gtggtagtaa tacatactac 180gcagactccg
tgaagggccg gttcaccatc tccagagaca attccaagaa cacgctgtat 240ctgcaaatga
acagcctgag agccgaggac acggccgtat attactgtgc gaaagatcga 300ctctctataa
caattcgccc acgctattac ggtctggacg tctggggcca agggaccacg 360gtcaccgtct
cct
373118124PRTArtificial SequenceSynthetic 118Glu Val Gln Leu Val Glu Ser
Gly Gly Gly Leu Val Gln Pro Gly Gly1 5 10
15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Asn
Phe Arg Asp Phe 20 25 30
Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45 Ser Ala Ile Ser
Gly Ser Gly Ser Asn Thr Tyr Tyr Ala Asp Ser Val 50 55
60 Lys Gly Arg Phe Thr Ile Ser Arg Asp
Asn Ser Lys Asn Thr Leu Tyr65 70 75
80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr
Tyr Cys 85 90 95
Ala Lys Asp Arg Leu Ser Ile Thr Ile Arg Pro Arg Tyr Tyr Gly Leu
100 105 110 Asp Val Trp Gly Gln
Gly Thr Thr Val Thr Val Ser 115 120
119322DNAArtificial SequenceSynthetic 119gacatccaga tgacccagtc
tccatcctcc ctgtctgcat ctgtaggaga cagagtcacc 60atcacttgcc gggcgagtca
ggacattagc aattatttag cctggtatca gcagaaacca 120gggaaagttc ctaagctcct
gatctatgct gcatccactt tgcaatcagg ggtcccatct 180cggttcagtg gcagtggatc
tgggacagat ttcactctca ccatcagcag cctgcagcct 240gaagatgttg caacttatta
ctgtcaaaaa tatgacagtg ccccgtacac ttttggccag 300gggaccaagc tggagatcaa
ac 322120107PRTArtificial
SequenceSynthetic 120Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala
Ser Val Gly1 5 10 15
Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Ser Asn Tyr
20 25 30 Leu Ala Trp Tyr Gln
Gln Lys Pro Gly Lys Val Pro Lys Leu Leu Ile 35 40
45 Tyr Ala Ala Ser Thr Leu Gln Ser Gly Val
Pro Ser Arg Phe Ser Gly 50 55 60
Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln
Pro65 70 75 80 Glu
Asp Val Ala Thr Tyr Tyr Cys Gln Lys Tyr Asp Ser Ala Pro Tyr
85 90 95 Thr Phe Gly Gln Gly Thr
Lys Leu Glu Ile Lys 100 105
121357DNAArtificial SequenceSynthetic 121caggtgcagc tggtggagtc tgggggaggc
ttggtacagc ctggggggtc cctgagactc 60tcctgtgtag cttctggatt cacccttaac
aactttgtca tgaactgggt ccgccaggtt 120ccagggaagg gactggagtg ggtctctttt
attagtgcta gtggtggtag tatatactac 180gcagactccg tgaagggccg gttcaccatc
tccagagaca cttccaagaa cacattatat 240ctgcaaatga acagcctgag agccgacgac
acggccgtct attactgtgc gaaatccccg 300tataactgga acccctttga ctattggggc
cagggaacca cggtcaccgt ctcctca 357122119PRTArtificial
SequenceSynthetic 122Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln
Pro Gly Gly1 5 10 15
Ser Leu Arg Leu Ser Cys Val Ala Ser Gly Phe Thr Leu Asn Asn Phe
20 25 30 Val Met Asn Trp Val
Arg Gln Val Pro Gly Lys Gly Leu Glu Trp Val 35 40
45 Ser Phe Ile Ser Ala Ser Gly Gly Ser Ile
Tyr Tyr Ala Asp Ser Val 50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Thr Ser Lys Asn Thr Leu
Tyr65 70 75 80 Leu
Gln Met Asn Ser Leu Arg Ala Asp Asp Thr Ala Val Tyr Tyr Cys
85 90 95 Ala Lys Ser Pro Tyr Asn
Trp Asn Pro Phe Asp Tyr Trp Gly Gln Gly 100
105 110 Thr Thr Val Thr Val Ser Ser 115
12324DNAArtificial SequenceSynthetic 123ggattcaccc
ttaacaactt tgtc
241248PRTArtificial SequenceSynthetic 124Gly Phe Thr Leu Asn Asn Phe Val1
5 12524DNAArtificial SequenceSynthetic
125attagtgcta gtggtggtag tata
241268PRTArtificial SequenceSynthetic 126Ile Ser Ala Ser Gly Gly Ser Ile1
5 12736DNAArtificial SequenceSynthetic
127gcgaaatccc cgtataactg gaaccccttt gactat
3612812PRTArtificial SequenceSynthetic 128Ala Lys Ser Pro Tyr Asn Trp Asn
Pro Phe Asp Tyr1 5 10
129327DNAArtificial SequenceSynthetic 129gacatccagt tgacccagtc tccagccacc
ctgtctgtgt ctccagggga acgagccacc 60ctctcctgca gggccagtct gagtgttagc
agcaaattag cctggtacca gcagacacct 120ggccaggctc ccagactcct catctatagt
gcctccaccc gggccactgg tatcccagtc 180aggttcagtg gcagtgggtc tgggacagag
ttcactctca ccatcagcag cctgcagtct 240gaagattttg cggtttatta ctgtcagcag
tataatcatt ggcctccgta cacttttggc 300caggggacca aggtggagat caaacga
327130109PRTArtificial
SequenceSynthetic 130Asp Ile Gln Leu Thr Gln Ser Pro Ala Thr Leu Ser Val
Ser Pro Gly1 5 10 15
Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Leu Ser Val Ser Ser Lys
20 25 30 Leu Ala Trp Tyr Gln
Gln Thr Pro Gly Gln Ala Pro Arg Leu Leu Ile 35 40
45 Tyr Ser Ala Ser Thr Arg Ala Thr Gly Ile
Pro Val Arg Phe Ser Gly 50 55 60
Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln
Ser65 70 75 80 Glu
Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Asn His Trp Pro Pro
85 90 95 Tyr Thr Phe Gly Gln Gly
Thr Lys Val Glu Ile Lys Arg 100 105
13118DNAArtificial SequenceSynthetic 131ctgagtgtta gcagcaaa
181326PRTArtificial
SequenceSynthetic 132Leu Ser Val Ser Ser Lys1 5
1339DNAArtificial SequenceSynthetic 133agtgcctcc
91343PRTArtificial SequenceSynthetic
134Ser Ala Ser1 13530DNAArtificial SequenceSynthetic
135cagcagtata atcattggcc tccgtacact
3013610PRTArtificial SequenceSynthetic 136Gln Gln Tyr Asn His Trp Pro Pro
Tyr Thr1 5 10 137357DNAArtificial
SequenceSynthetic 137gaggtgcagc tggtggagtc tgggggaggc ttggtacagc
ctggggggtc cctgagactc 60tcctgtgtag cttctggatt cacccttaac aactttgtca
tgaactgggt ccgccaggtt 120ccagggaagg gactggagtg ggtctctttt attagtgcta
gtggtggtag tatatactac 180gcagactccg tgaagggccg gttcaccatc tccagagaca
cttccaagaa cacattatat 240ctgcaaatga acagcctgag agccgacgac acggccgtct
attactgtgc gaaatccccg 300tataactgga acccctttga ctattggggc cagggaaccc
tggtcaccgt ctcctca 357138119PRTArtificial SequenceSynthetic 138Glu
Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly1
5 10 15 Ser Leu Arg Leu Ser Cys
Val Ala Ser Gly Phe Thr Leu Asn Asn Phe 20 25
30 Val Met Asn Trp Val Arg Gln Val Pro Gly Lys
Gly Leu Glu Trp Val 35 40 45
Ser Phe Ile Ser Ala Ser Gly Gly Ser Ile Tyr Tyr Ala Asp Ser Val
50 55 60 Lys Gly Arg
Phe Thr Ile Ser Arg Asp Thr Ser Lys Asn Thr Leu Tyr65 70
75 80 Leu Gln Met Asn Ser Leu Arg Ala
Asp Asp Thr Ala Val Tyr Tyr Cys 85 90
95 Ala Lys Ser Pro Tyr Asn Trp Asn Pro Phe Asp Tyr Trp
Gly Gln Gly 100 105 110
Thr Leu Val Thr Val Ser Ser 115
139324DNAArtificial SequenceSynthetic 139gaaatagtga tgacgcagtc tccagccacc
ctgtctgtgt ctccagggga acgagccacc 60ctctcctgca gggccagtct gagtgttagc
agcaaattag cctggtacca gcagacacct 120ggccaggctc ccagactcct catctatagt
gcctccaccc gggccactgg tatcccagtc 180aggttcagtg gcagtgggtc tgggacagag
ttcactctca ccatcagcag cctgcagtct 240gaagattttg cggtttatta ctgtcagcag
tataatcatt ggcctccgta cacttttggc 300caggggacca agctggagat caaa
324140108PRTArtificial
SequenceSynthetic 140Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Val
Ser Pro Gly1 5 10 15
Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Leu Ser Val Ser Ser Lys
20 25 30 Leu Ala Trp Tyr Gln
Gln Thr Pro Gly Gln Ala Pro Arg Leu Leu Ile 35 40
45 Tyr Ser Ala Ser Thr Arg Ala Thr Gly Ile
Pro Val Arg Phe Ser Gly 50 55 60
Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln
Ser65 70 75 80 Glu
Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Asn His Trp Pro Pro
85 90 95 Tyr Thr Phe Gly Gln Gly
Thr Lys Leu Glu Ile Lys 100 105
141357DNAArtificial SequenceSynthetic 141gaggtgcagc tggtggagtc tgggggaggc
ttggtacagc ctggggggtc cctgagactc 60tcctgtgcag cctctggatt cacccttaac
aactttgtca tgagctgggt ccgccaggct 120ccagggaagg ggctggagtg ggtctcagct
attagtgcta gtggtggtag tatatactac 180gcagactccg tgaagggccg gttcaccatc
tccagagaca attccaagaa cacgctgtat 240ctgcaaatga acagcctgag agccgaggac
acggccgtat attactgtgc gaaatccccg 300tataactgga acccctttga ctattggggc
cagggaaccc tggtcaccgt ctcctca 357142119PRTArtificial
SequenceSynthetic 142Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln
Pro Gly Gly1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Leu Asn Asn Phe
20 25 30 Val Met Ser Trp Val
Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40
45 Ser Ala Ile Ser Ala Ser Gly Gly Ser Ile
Tyr Tyr Ala Asp Ser Val 50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu
Tyr65 70 75 80 Leu
Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95 Ala Lys Ser Pro Tyr Asn
Trp Asn Pro Phe Asp Tyr Trp Gly Gln Gly 100
105 110 Thr Leu Val Thr Val Ser Ser 115
143325DNAArtificial SequenceSynthetic 143gaaatagtga
tgacgcagtc tccagccacc ctgtctgtgt ctccagggga aagagccacc 60ctctcctgca
gggccagtct gagtgttagc agcaaattag cctggtacca gcagaaacct 120ggccaggctc
ccaggctcct catctatagt gcctccacca gggccactgg tatcccagcc 180aggttcagtg
gcagtgggtc tgggacagag ttcactctca ccatcagcag cctgcagtct 240gaagattttg
cagtttatta ctgtcagcag tataatcatt ggcctccgta cacttttggc 300caggggacca
agctggagat caaac
325144108PRTArtificial SequenceSynthetic 144Glu Ile Val Met Thr Gln Ser
Pro Ala Thr Leu Ser Val Ser Pro Gly1 5 10
15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Leu Ser
Val Ser Ser Lys 20 25 30
Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile
35 40 45 Tyr Ser Ala Ser
Thr Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly 50 55
60 Ser Gly Ser Gly Thr Glu Phe Thr Leu
Thr Ile Ser Ser Leu Gln Ser65 70 75
80 Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Asn His Trp
Pro Pro 85 90 95
Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100
105 145375DNAArtificial SequenceSynthetic 145caggtgcagc
tggtggagtc tgggggaggc ttggaacagc cgggggggtc cctgagactc 60tcctgtgcag
gctctggatt cacctttaga gactatgcca tgacctgggt ccgccaggct 120ccagggaagg
gactggagtg ggtctcatct attagtggtt ccggtggtaa cacatactac 180gcagactccg
tgaagggccg gttcaccatc tccagagaca attccaagaa cacgctgtat 240ctgcaaatga
acagcctgag agccgaggac acggccgtat attactgtgc gaaagatcga 300ctctctataa
caattcgccc acgctattat ggtttggacg tctggggcca agggtccacg 360gtcaccgtct
cctca
375146125PRTArtificial SequenceSynthetic 146Gln Val Gln Leu Val Glu Ser
Gly Gly Gly Leu Glu Gln Pro Gly Gly1 5 10
15 Ser Leu Arg Leu Ser Cys Ala Gly Ser Gly Phe Thr
Phe Arg Asp Tyr 20 25 30
Ala Met Thr Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45 Ser Ser Ile Ser
Gly Ser Gly Gly Asn Thr Tyr Tyr Ala Asp Ser Val 50 55
60 Lys Gly Arg Phe Thr Ile Ser Arg Asp
Asn Ser Lys Asn Thr Leu Tyr65 70 75
80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr
Tyr Cys 85 90 95
Ala Lys Asp Arg Leu Ser Ile Thr Ile Arg Pro Arg Tyr Tyr Gly Leu
100 105 110 Asp Val Trp Gly Gln
Gly Ser Thr Val Thr Val Ser Ser 115 120
125 14724DNAArtificial SequenceSynthetic 147ggattcacct ttagagacta
tgcc 241488PRTArtificial
SequenceSynthetic 148Gly Phe Thr Phe Arg Asp Tyr Ala1 5
14924DNAArtificial SequenceSynthetic 149attagtggtt ccggtggtaa
caca 241508PRTArtificial
SequenceSynthetic 150Ile Ser Gly Ser Gly Gly Asn Thr1 5
15154DNAArtificial SequenceSynthetic 151gcgaaagatc gactctctat
aacaattcgc ccacgctatt atggtttgga cgtc 5415218PRTArtificial
SequenceSynthetic 152Ala Lys Asp Arg Leu Ser Ile Thr Ile Arg Pro Arg Tyr
Tyr Gly Leu1 5 10 15
Asp Val153339DNAArtificial SequenceSynthetic 153gacatcgtgt tgacccagtc
tccactctcc ctgcccgtca cccctggaga gccggcctcc 60atctcctgca ggtctagtca
gagcctcctg tatagtattg gatacaacta tttggattgg 120tacctgcaga agtcagggca
gtctccacag ctccttatct atttgggttc taatcgggcc 180tccggggtcc ctgacaggtt
cagtggcagt ggatcaggca cagattttac actgaaaatc 240agcagagtgg aggctgagga
tgttgggttt tattactgca tgcaagctct acaaactccg 300tacacttttg gccaggggac
caagctggag atcaaacga 339154113PRTArtificial
SequenceSynthetic 154Asp Ile Val Leu Thr Gln Ser Pro Leu Ser Leu Pro Val
Thr Pro Gly1 5 10 15
Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Leu Tyr Ser
20 25 30 Ile Gly Tyr Asn Tyr
Leu Asp Trp Tyr Leu Gln Lys Ser Gly Gln Ser 35 40
45 Pro Gln Leu Leu Ile Tyr Leu Gly Ser Asn
Arg Ala Ser Gly Val Pro 50 55 60
Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys
Ile65 70 75 80 Ser
Arg Val Glu Ala Glu Asp Val Gly Phe Tyr Tyr Cys Met Gln Ala
85 90 95 Leu Gln Thr Pro Tyr Thr
Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100
105 110 Arg15533DNAArtificial SequenceSynthetic
155cagagcctcc tgtatagtat tggatacaac tat
3315611PRTArtificial SequenceSynthetic 156Gln Ser Leu Leu Tyr Ser Ile Gly
Tyr Asn Tyr1 5 10 1579DNAArtificial
SequenceSynthetic 157ttgggttct
91583PRTArtificial SequenceSynthetic 158Leu Gly Ser1
15927DNAArtificial SequenceSynthetic 159atgcaagctc tacaaactcc
gtacact 271609PRTArtificial
SequenceSynthetic 160Met Gln Ala Leu Gln Thr Pro Tyr Thr1 5
161372DNAArtificial SequenceSynthetic 161gaggtgcagc
tggtggagtc tgggggaggc ttggaacagc cgggggggtc cctgagactc 60tcctgtgcag
gctctggatt cacctttaga gactatgcca tgacctgggt ccgccaggct 120ccagggaagg
gactggagtg ggtctcatct attagtggtt ccggtggtaa cacatactac 180gcagactccg
tgaagggccg gttcaccatc tccagagaca attccaagaa cacgctgtat 240ctgcaaatga
acagcctgag agccgaggac acggccgtat attactgtgc gaaagatcga 300ctctctataa
caattcgccc acgctattat ggtttggacg tctggggcca agggaccacg 360gtcaccgtct
cc
372162124PRTArtificial SequenceSynthetic 162Glu Val Gln Leu Val Glu Ser
Gly Gly Gly Leu Glu Gln Pro Gly Gly1 5 10
15 Ser Leu Arg Leu Ser Cys Ala Gly Ser Gly Phe Thr
Phe Arg Asp Tyr 20 25 30
Ala Met Thr Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45 Ser Ser Ile Ser
Gly Ser Gly Gly Asn Thr Tyr Tyr Ala Asp Ser Val 50 55
60 Lys Gly Arg Phe Thr Ile Ser Arg Asp
Asn Ser Lys Asn Thr Leu Tyr65 70 75
80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr
Tyr Cys 85 90 95
Ala Lys Asp Arg Leu Ser Ile Thr Ile Arg Pro Arg Tyr Tyr Gly Leu
100 105 110 Asp Val Trp Gly Gln
Gly Thr Thr Val Thr Val Ser 115 120
163336DNAArtificial SequenceSynthetic 163gacatcgtga tgacccagtc
tccactctcc ctgcccgtca cccctggaga gccggcctcc 60atctcctgca ggtctagtca
gagcctcctg tatagtattg gatacaacta tttggattgg 120tacctgcaga agtcagggca
gtctccacag ctccttatct atttgggttc taatcgggcc 180tccggggtcc ctgacaggtt
cagtggcagt ggatcaggca cagattttac actgaaaatc 240agcagagtgg aggctgagga
tgttgggttt tattactgca tgcaagctct acaaactccg 300tacacttttg gccaggggac
caagctggag atcaaa 336164112PRTArtificial
SequenceSynthetic 164Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val
Thr Pro Gly1 5 10 15
Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Leu Tyr Ser
20 25 30 Ile Gly Tyr Asn Tyr
Leu Asp Trp Tyr Leu Gln Lys Ser Gly Gln Ser 35 40
45 Pro Gln Leu Leu Ile Tyr Leu Gly Ser Asn
Arg Ala Ser Gly Val Pro 50 55 60
Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys
Ile65 70 75 80 Ser
Arg Val Glu Ala Glu Asp Val Gly Phe Tyr Tyr Cys Met Gln Ala
85 90 95 Leu Gln Thr Pro Tyr Thr
Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100
105 110 165373DNAArtificial SequenceSynthetic
165gaggtgcagc tggtggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc
60tcctgtgcag cctctggatt cacctttaga gactatgcca tgagctgggt ccgccaggct
120ccagggaagg ggctggagtg ggtctcagct attagtggtt ccggtggtaa cacatactac
180gcagactccg tgaagggccg gttcaccatc tccagagaca attccaagaa cacgctgtat
240ctgcaaatga acagcctgag agccgaggac acggccgtat attactgtgc gaaagatcga
300ctctctataa caattcgccc acgctattat ggtttggacg tctggggcca agggaccacg
360gtcaccgtct cct
373166124PRTArtificial SequenceSynthetic 166Glu Val Gln Leu Val Glu Ser
Gly Gly Gly Leu Val Gln Pro Gly Gly1 5 10
15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr
Phe Arg Asp Tyr 20 25 30
Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45 Ser Ala Ile Ser
Gly Ser Gly Gly Asn Thr Tyr Tyr Ala Asp Ser Val 50 55
60 Lys Gly Arg Phe Thr Ile Ser Arg Asp
Asn Ser Lys Asn Thr Leu Tyr65 70 75
80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr
Tyr Cys 85 90 95
Ala Lys Asp Arg Leu Ser Ile Thr Ile Arg Pro Arg Tyr Tyr Gly Leu
100 105 110 Asp Val Trp Gly Gln
Gly Thr Thr Val Thr Val Ser 115 120
167337DNAArtificial SequenceSynthetic 167gatattgtga tgactcagtc
tccactctcc ctgcccgtca cccctggaga gccggcctcc 60atctcctgca ggtctagtca
gagcctcctg tatagtattg gatacaacta tttggattgg 120tacctgcaga agccagggca
gtctccacag ctcctgatct atttgggttc taatcgggcc 180tccggggtcc ctgacaggtt
cagtggcagt ggatcaggca cagattttac actgaaaatc 240agcagagtgg aggctgagga
tgttggggtt tattactgca tgcaagctct acaaactccg 300tacacttttg gccaggggac
caagctggag atcaaac 337168112PRTArtificial
SequenceSynthetic 168Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val
Thr Pro Gly1 5 10 15
Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Leu Tyr Ser
20 25 30 Ile Gly Tyr Asn Tyr
Leu Asp Trp Tyr Leu Gln Lys Pro Gly Gln Ser 35 40
45 Pro Gln Leu Leu Ile Tyr Leu Gly Ser Asn
Arg Ala Ser Gly Val Pro 50 55 60
Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys
Ile65 70 75 80 Ser
Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Met Gln Ala
85 90 95 Leu Gln Thr Pro Tyr Thr
Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100
105 110 169375DNAArtificial SequenceSynthetic
169caggtgcagc tggtggagtc tgggggagtc ttggagcagc ctggggggtc cctgagactc
60tcctgtacag cctctggatt cacctttaga gactatgcca tgacctgggt ccgccaggct
120ccagggaagg ggctggagtg ggtctcatct attagtggta gtggtggtaa tacatactac
180gcagactccg tgaggggccg gttcaccatc tccagagaca actccaacca cacgctgtat
240ctgcaaatga acagcctgag agccgaagac acggccgtat attactgtgc gaaagatcga
300ctctccataa caattcgccc acgctattac ggtttggacg tctggggcca agggtccacg
360gtcaccgtct cctca
375170125PRTArtificial SequenceSynthetic 170Gln Val Gln Leu Val Glu Ser
Gly Gly Val Leu Glu Gln Pro Gly Gly1 5 10
15 Ser Leu Arg Leu Ser Cys Thr Ala Ser Gly Phe Thr
Phe Arg Asp Tyr 20 25 30
Ala Met Thr Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45 Ser Ser Ile Ser
Gly Ser Gly Gly Asn Thr Tyr Tyr Ala Asp Ser Val 50 55
60 Arg Gly Arg Phe Thr Ile Ser Arg Asp
Asn Ser Asn His Thr Leu Tyr65 70 75
80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr
Tyr Cys 85 90 95
Ala Lys Asp Arg Leu Ser Ile Thr Ile Arg Pro Arg Tyr Tyr Gly Leu
100 105 110 Asp Val Trp Gly Gln
Gly Ser Thr Val Thr Val Ser Ser 115 120
125 17124DNAArtificial SequenceSynthetic 171ggattcacct ttagagacta
tgcc 241728PRTArtificial
SequenceSynthetic 172Gly Phe Thr Phe Arg Asp Tyr Ala1 5
17324DNAArtificial SequenceSynthetic 173attagtggta gtggtggtaa
taca 241748PRTArtificial
SequenceSynthetic 174Ile Ser Gly Ser Gly Gly Asn Thr1 5
17554DNAArtificial SequenceSynthetic 175gcgaaagatc gactctccat
aacaattcgc ccacgctatt acggtttgga cgtc 5417618PRTArtificial
SequenceSynthetic 176Ala Lys Asp Arg Leu Ser Ile Thr Ile Arg Pro Arg Tyr
Tyr Gly Leu1 5 10 15
Asp Val177324DNAArtificial SequenceSynthetic 177gatattgtga tgacccagtc
tccatcctcc ctgtctgcat ctgtaggaga cagagtcacc 60attacttgcc gggcgagtca
ggacattagc aattattttg cctggtatca gcagaagcca 120gggaaagttc ctaaactcct
gatctttgct gcatccactt tgcatccagg ggtcccatct 180cggttcagtg gcagtggatc
tgggacagat ttcactctca ccattagtag cctgcagcct 240gaagatgttg caacttatta
ctgtcaaaag tataacagtg ccccgtacac ttttggccag 300gggaccaagg tggaaatcaa
acga 324178108PRTArtificial
SequenceSynthetic 178Asp Ile Val Met Thr Gln Ser Pro Ser Ser Leu Ser Ala
Ser Val Gly1 5 10 15
Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Ser Asn Tyr
20 25 30 Phe Ala Trp Tyr Gln
Gln Lys Pro Gly Lys Val Pro Lys Leu Leu Ile 35 40
45 Phe Ala Ala Ser Thr Leu His Pro Gly Val
Pro Ser Arg Phe Ser Gly 50 55 60
Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln
Pro65 70 75 80 Glu
Asp Val Ala Thr Tyr Tyr Cys Gln Lys Tyr Asn Ser Ala Pro Tyr
85 90 95 Thr Phe Gly Gln Gly Thr
Lys Val Glu Ile Lys Arg 100 105
17918DNAArtificial SequenceSynthetic 179caggacatta gcaattat
181806PRTArtificial SequenceSynthetic
180Gln Asp Ile Ser Asn Tyr1 5 1819DNAArtificial
SequenceSynthetic 181gctgcatcc
91823PRTArtificial SequenceSynthetic 182Ala Ala Ser1
18327DNAArtificial SequenceSynthetic 183caaaagtata acagtgcccc
gtacact 271849PRTArtificial
SequenceSynthetic 184Gln Lys Tyr Asn Ser Ala Pro Tyr Thr1 5
185372DNAArtificial SequenceSynthetic 185gaggtgcagc
tggtggagtc tgggggagtc ttggagcagc ctggggggtc cctgagactc 60tcctgtacag
cctctggatt cacctttaga gactatgcca tgacctgggt ccgccaggct 120ccagggaagg
ggctggagtg ggtctcatct attagtggta gtggtggtaa tacatactac 180gcagactccg
tgaggggccg gttcaccatc tccagagaca actccaacca cacgctgtat 240ctgcaaatga
acagcctgag agccgaagac acggccgtat attactgtgc gaaagatcga 300ctctccataa
caattcgccc acgctattac ggtttggacg tctggggcca agggaccacg 360gtcaccgtct
cc
372186124PRTArtificial SequenceSynthetic 186Glu Val Gln Leu Val Glu Ser
Gly Gly Val Leu Glu Gln Pro Gly Gly1 5 10
15 Ser Leu Arg Leu Ser Cys Thr Ala Ser Gly Phe Thr
Phe Arg Asp Tyr 20 25 30
Ala Met Thr Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45 Ser Ser Ile Ser
Gly Ser Gly Gly Asn Thr Tyr Tyr Ala Asp Ser Val 50 55
60 Arg Gly Arg Phe Thr Ile Ser Arg Asp
Asn Ser Asn His Thr Leu Tyr65 70 75
80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr
Tyr Cys 85 90 95
Ala Lys Asp Arg Leu Ser Ile Thr Ile Arg Pro Arg Tyr Tyr Gly Leu
100 105 110 Asp Val Trp Gly Gln
Gly Thr Thr Val Thr Val Ser 115 120
187321DNAArtificial SequenceSynthetic 187gacatccaga tgacccagtc
tccatcctcc ctgtctgcat ctgtaggaga cagagtcacc 60attacttgcc gggcgagtca
ggacattagc aattattttg cctggtatca gcagaagcca 120gggaaagttc ctaaactcct
gatctttgct gcatccactt tgcatccagg ggtcccatct 180cggttcagtg gcagtggatc
tgggacagat ttcactctca ccattagtag cctgcagcct 240gaagatgttg caacttatta
ctgtcaaaag tataacagtg ccccgtacac ttttggccag 300gggaccaagc tggagatcaa a
321188107PRTArtificial
SequenceSynthetic 188Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala
Ser Val Gly1 5 10 15
Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Ser Asn Tyr
20 25 30 Phe Ala Trp Tyr Gln
Gln Lys Pro Gly Lys Val Pro Lys Leu Leu Ile 35 40
45 Phe Ala Ala Ser Thr Leu His Pro Gly Val
Pro Ser Arg Phe Ser Gly 50 55 60
Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln
Pro65 70 75 80 Glu
Asp Val Ala Thr Tyr Tyr Cys Gln Lys Tyr Asn Ser Ala Pro Tyr
85 90 95 Thr Phe Gly Gln Gly Thr
Lys Leu Glu Ile Lys 100 105
189373DNAArtificial SequenceSynthetic 189gaggtgcagc tggtggagtc tgggggaggc
ttggtacagc ctggggggtc cctgagactc 60tcctgtgcag cctctggatt cacctttaga
gactatgcca tgagctgggt ccgccaggct 120ccagggaagg ggctggagtg ggtctcagct
attagtggta gtggtggtaa tacatactac 180gcagactccg tgaagggccg gttcaccatc
tccagagaca attccaagaa cacgctgtat 240ctgcaaatga acagcctgag agccgaggac
acggccgtat attactgtgc gaaagatcga 300ctctccataa caattcgccc acgctattac
ggtttggacg tctggggcca agggaccacg 360gtcaccgtct cct
373190124PRTArtificial
SequenceSynthetic 190Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln
Pro Gly Gly1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Arg Asp Tyr
20 25 30 Ala Met Ser Trp Val
Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40
45 Ser Ala Ile Ser Gly Ser Gly Gly Asn Thr
Tyr Tyr Ala Asp Ser Val 50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu
Tyr65 70 75 80 Leu
Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95 Ala Lys Asp Arg Leu Ser
Ile Thr Ile Arg Pro Arg Tyr Tyr Gly Leu 100
105 110 Asp Val Trp Gly Gln Gly Thr Thr Val Thr
Val Ser 115 120
191322DNAArtificial SequenceSynthetic 191gacatccaga tgacccagtc tccatcctcc
ctgtctgcat ctgtaggaga cagagtcacc 60atcacttgcc gggcgagtca ggacattagc
aattatttag cctggtatca gcagaaacca 120gggaaagttc ctaagctcct gatctatgct
gcatccactt tgcaatcagg ggtcccatct 180cggttcagtg gcagtggatc tgggacagat
ttcactctca ccatcagcag cctgcagcct 240gaagatgttg caacttatta ctgtcaaaag
tataacagtg ccccgtacac ttttggccag 300gggaccaagc tggagatcaa ac
322192107PRTArtificial
SequenceSynthetic 192Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala
Ser Val Gly1 5 10 15
Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Ser Asn Tyr
20 25 30 Leu Ala Trp Tyr Gln
Gln Lys Pro Gly Lys Val Pro Lys Leu Leu Ile 35 40
45 Tyr Ala Ala Ser Thr Leu Gln Ser Gly Val
Pro Ser Arg Phe Ser Gly 50 55 60
Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln
Pro65 70 75 80 Glu
Asp Val Ala Thr Tyr Tyr Cys Gln Lys Tyr Asn Ser Ala Pro Tyr
85 90 95 Thr Phe Gly Gln Gly Thr
Lys Leu Glu Ile Lys 100 105
193355DNAArtificial SequenceSynthetic 193gaagtgcacc tggtggaatc tgggggaggc
ttggtacagc ctggcaggtc cctgagactc 60tcctgtgagg cctctggatt cacctttgat
gattatgcca tgcactgggt ccggcaagct 120ccggggaagg gcctggaatg ggtctcaggt
cttagtcgga caagtgtcag tataggctat 180gcggactctg tgaagggccg attcaccatc
tccagagaca acgccaagaa ctccctttat 240ttggaaatga acagtctgag acctgaggac
acggccttat attactgtgc aaaatggggg 300acccgggggt attttgacta ctggggccag
ggaaccctgg tcaccgtctc ctcag 355194118PRTArtificial
SequenceSynthetic 194Glu Val His Leu Val Glu Ser Gly Gly Gly Leu Val Gln
Pro Gly Arg1 5 10 15
Ser Leu Arg Leu Ser Cys Glu Ala Ser Gly Phe Thr Phe Asp Asp Tyr
20 25 30 Ala Met His Trp Val
Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40
45 Ser Gly Leu Ser Arg Thr Ser Val Ser Ile
Gly Tyr Ala Asp Ser Val 50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu
Tyr65 70 75 80 Leu
Glu Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Leu Tyr Tyr Cys
85 90 95 Ala Lys Trp Gly Thr Arg
Gly Tyr Phe Asp Tyr Trp Gly Gln Gly Thr 100
105 110 Leu Val Thr Val Ser Ser 115
19524DNAArtificial SequenceSynthetic 195ggattcacct ttgatgatta tgcc
241968PRTArtificial
SequenceSynthetic 196Gly Phe Thr Phe Asp Asp Tyr Ala1 5
19724DNAArtificial SequenceSynthetic 197cttagtcgga caagtgtcag
tata 241988PRTArtificial
SequenceSynthetic 198Leu Ser Arg Thr Ser Val Ser Ile1 5
19933DNAArtificial SequenceSynthetic 199gcaaaatggg ggacccgggg
gtattttgac tac 3320011PRTArtificial
SequenceSynthetic 200Ala Lys Trp Gly Thr Arg Gly Tyr Phe Asp Tyr1
5 10 201322DNAArtificial SequenceSynthetic
201gacatccaga tgacccagtc tccatcttcc gtgtctgcat ctgtgggaga cagagtcacc
60atcacttgtc gggcgagtca ggatattagt atttggttag cctggtatca gcagagtcca
120gggaaagccc ctaaactcct gatcaatgtt gcatcccgtt tgcaaagtgg ggtcccatca
180aggttcagcg gcagtggatc tgggacagat ttcactctca ccatcaacag tctgcagcct
240gaagattttg taacttacta ttgtcaacag gctaacagtt tcccgatcac cttcggccaa
300gggacacgac tggcgaccaa ac
322202107PRTArtificial SequenceSynthetic 202Asp Ile Gln Met Thr Gln Ser
Pro Ser Ser Val Ser Ala Ser Val Gly1 5 10
15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp
Ile Ser Ile Trp 20 25 30
Leu Ala Trp Tyr Gln Gln Ser Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45 Asn Val Ala Ser
Arg Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55
60 Ser Gly Ser Gly Thr Asp Phe Thr Leu
Thr Ile Asn Ser Leu Gln Pro65 70 75
80 Glu Asp Phe Val Thr Tyr Tyr Cys Gln Gln Ala Asn Ser Phe
Pro Ile 85 90 95
Thr Phe Gly Gln Gly Thr Arg Leu Ala Thr Lys 100
105 20318DNAArtificial SequenceSynthetic 203caggatatta gtatttgg
182046PRTArtificial
SequenceSynthetic 204Gln Asp Ile Ser Ile Trp1 5
2059DNAArtificial SequenceSynthetic 205gttgcatcc
92063PRTArtificial SequenceSynthetic
206Val Ala Ser1 20727DNAArtificial SequenceSynthetic
207caacaggcta acagtttccc gatcacc
272089PRTArtificial SequenceSynthetic 208Gln Gln Ala Asn Ser Phe Pro Ile
Thr1 5 209355DNAArtificial
SequenceSynthetic 209gaggtgcagc tggtggagtc tgggggaggc ttggtacagc
ctggcaggtc cctgagactc 60tcctgtgagg cctctggatt cacctttgat gattatgcca
tgcactgggt ccggcaagct 120ccggggaagg gcctggaatg ggtctcaggt cttagtcgga
caagtgtcag tataggctat 180gcggactctg tgaagggccg attcaccatc tccagagaca
acgccaagaa ctccctttat 240ttggaaatga acagtctgag acctgaggac acggccttat
attactgtgc aaaatggggg 300acccgggggt attttgacta ctggggccag ggaaccctgg
tcaccgtctc ctcag 355210118PRTArtificial SequenceSynthetic 210Glu
Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg1
5 10 15 Ser Leu Arg Leu Ser Cys
Glu Ala Ser Gly Phe Thr Phe Asp Asp Tyr 20 25
30 Ala Met His Trp Val Arg Gln Ala Pro Gly Lys
Gly Leu Glu Trp Val 35 40 45
Ser Gly Leu Ser Arg Thr Ser Val Ser Ile Gly Tyr Ala Asp Ser Val
50 55 60 Lys Gly Arg
Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr65 70
75 80 Leu Glu Met Asn Ser Leu Arg Pro
Glu Asp Thr Ala Leu Tyr Tyr Cys 85 90
95 Ala Lys Trp Gly Thr Arg Gly Tyr Phe Asp Tyr Trp Gly
Gln Gly Thr 100 105 110
Leu Val Thr Val Ser Ser 115 211322DNAArtificial
SequenceSynthetic 211gacatccaga tgacccagtc tccatcttcc gtgtctgcat
ctgtgggaga cagagtcacc 60atcacttgtc gggcgagtca ggatattagt atttggttag
cctggtatca gcagagtcca 120gggaaagccc ctaaactcct gatcaatgtt gcatcccgtt
tgcaaagtgg ggtcccatca 180aggttcagcg gcagtggatc tgggacagat ttcactctca
ccatcaacag tctgcagcct 240gaagattttg taacttacta ttgtcaacag gctaacagtt
tcccgatcac cttcggccaa 300gggacacgac tggagattaa ac
322212107PRTArtificial SequenceSynthetic 212Asp
Ile Gln Met Thr Gln Ser Pro Ser Ser Val Ser Ala Ser Val Gly1
5 10 15 Asp Arg Val Thr Ile Thr
Cys Arg Ala Ser Gln Asp Ile Ser Ile Trp 20 25
30 Leu Ala Trp Tyr Gln Gln Ser Pro Gly Lys Ala
Pro Lys Leu Leu Ile 35 40 45
Asn Val Ala Ser Arg Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly
50 55 60 Ser Gly Ser
Gly Thr Asp Phe Thr Leu Thr Ile Asn Ser Leu Gln Pro65 70
75 80 Glu Asp Phe Val Thr Tyr Tyr Cys
Gln Gln Ala Asn Ser Phe Pro Ile 85 90
95 Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys
100 105 213355DNAArtificial SequenceSynthetic
213gaagtgcagc tggtggagtc tgggggaggc ttggtacagc ctggcaggtc cctgagactc
60tcctgtgcag cctctggatt cacctttgat gattatgcca tgcactgggt ccggcaagct
120ccagggaagg gcctggagtg ggtctcaggt cttagtcgga caagtgtcag tataggctat
180gcggactctg tgaagggccg attcaccatc tccagagaca acgccaagaa ctccctgtat
240ctgcaaatga acagtctgag agctgaggac acggccttgt attactgtgc aaaatggggg
300acccgggggt attttgacta ctggggccaa ggaaccctgg tcaccgtctc ctcag
355214118PRTArtificial SequenceSynthetic 214Glu Val Gln Leu Val Glu Ser
Gly Gly Gly Leu Val Gln Pro Gly Arg1 5 10
15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr
Phe Asp Asp Tyr 20 25 30
Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45 Ser Gly Leu Ser
Arg Thr Ser Val Ser Ile Gly Tyr Ala Asp Ser Val 50 55
60 Lys Gly Arg Phe Thr Ile Ser Arg Asp
Asn Ala Lys Asn Ser Leu Tyr65 70 75
80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Leu Tyr
Tyr Cys 85 90 95
Ala Lys Trp Gly Thr Arg Gly Tyr Phe Asp Tyr Trp Gly Gln Gly Thr
100 105 110 Leu Val Thr Val Ser
Ser 115 215322DNAArtificial SequenceSynthetic
215gacatccaga tgacccagtc tccatcttcc gtgtctgcat ctgtaggaga cagagtcacc
60atcacttgtc gggcgagtca ggatattagt atttggttag cctggtatca gcagaaacca
120gggaaagccc ctaagctcct gatctatgtt gcatccagtt tgcaaagtgg ggtcccatca
180aggttcagcg gcagtggatc tgggacagat ttcactctca ccatcagcag cctgcagcct
240gaagattttg caacttacta ttgtcaacag gctaacagtt tcccgatcac cttcggccaa
300gggacacgac tggagattaa ac
322216107PRTArtificial SequenceSynthetic 216Asp Ile Gln Met Thr Gln Ser
Pro Ser Ser Val Ser Ala Ser Val Gly1 5 10
15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp
Ile Ser Ile Trp 20 25 30
Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45 Tyr Val Ala Ser
Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55
60 Ser Gly Ser Gly Thr Asp Phe Thr Leu
Thr Ile Ser Ser Leu Gln Pro65 70 75
80 Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ala Asn Ser Phe
Pro Ile 85 90 95
Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys 100
105 217363DNAArtificial SequenceSynthetic 217gaggtgcagc
tgttggagtc tgggggaggc ttgctacagc cgggggggtc cctgagactc 60tcctgtgcag
cctctggaat cacctttagc acctatgcca tgagctgggt ccgtcaggct 120ccagggaggg
ggctggagtg ggtctcagct attagtggta gtggtgatag cacatcctac 180gcagactccg
tgaagggccg gttcaccagc tccagagaca attccaagaa cacgctgtat 240ctgcaaatga
acagcctgag agccgaggac acggccgtat attactgtgc gaaagtcata 300gcagctcgtc
ctcactggaa cttcgatctc tggggccgtg gcaccctggt cactgtctcc 360tca
363218121PRTArtificial SequenceSynthetic 218Glu Val Gln Leu Leu Glu Ser
Gly Gly Gly Leu Leu Gln Pro Gly Gly1 5 10
15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Ile Thr
Phe Ser Thr Tyr 20 25 30
Ala Met Ser Trp Val Arg Gln Ala Pro Gly Arg Gly Leu Glu Trp Val
35 40 45 Ser Ala Ile Ser
Gly Ser Gly Asp Ser Thr Ser Tyr Ala Asp Ser Val 50 55
60 Lys Gly Arg Phe Thr Ser Ser Arg Asp
Asn Ser Lys Asn Thr Leu Tyr65 70 75
80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr
Tyr Cys 85 90 95
Ala Lys Val Ile Ala Ala Arg Pro His Trp Asn Phe Asp Leu Trp Gly
100 105 110 Arg Gly Thr Leu Val
Thr Val Ser Ser 115 120 21924DNAArtificial
SequenceSynthetic 219ggaatcacct ttagcaccta tgcc
242208PRTArtificial SequenceSynthetic 220Gly Ile Thr Phe
Ser Thr Tyr Ala1 5 22124DNAArtificial
SequenceSynthetic 221attagtggta gtggtgatag caca
242228PRTArtificial SequenceSynthetic 222Ile Ser Gly Ser
Gly Asp Ser Thr1 5 22342DNAArtificial
SequenceSynthetic 223gcgaaagtca tagcagctcg tcctcactgg aacttcgatc tc
4222414PRTArtificial SequenceSynthetic 224Ala Lys Val
Ile Ala Ala Arg Pro His Trp Asn Phe Asp Leu1 5
10 225324DNAArtificial SequenceSynthetic
225gaaattgtgt tgacacagtc tccagccacc ctgtctttgt ctccagggga aagagccacc
60ctctcctgca gggccagtca gagtgttagt agatatttag cctggtatca acagaaacct
120ggccaggctc ccaggctcct catctatgat gcatccaaca gggccactgg catcccagcc
180aggttcagtg gcagtgggtc tgggacagac ttcactctca ccatcagcag cctagagcct
240gaagattttg gagtttatta ctgtcagcag cgtagtgact ggccgctcac tttcggcgga
300gggaccaagg tggagatcaa acgg
324226107PRTArtificial SequenceSynthetic 226Glu Ile Val Leu Thr Gln Ser
Pro Ala Thr Leu Ser Leu Ser Pro Gly1 5 10
15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser
Val Ser Arg Tyr 20 25 30
Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile
35 40 45 Tyr Asp Ala Ser
Asn Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly 50 55
60 Ser Gly Ser Gly Thr Asp Phe Thr Leu
Thr Ile Ser Ser Leu Glu Pro65 70 75
80 Glu Asp Phe Gly Val Tyr Tyr Cys Gln Gln Arg Ser Asp Trp
Pro Leu 85 90 95
Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100
105 22718DNAArtificial SequenceSynthetic 227cagagtgtta gtagatat
182286PRTArtificial
SequenceSynthetic 228Gln Ser Val Ser Arg Tyr1 5
2299DNAArtificial SequenceSynthetic 229gatgcatcc
92303PRTArtificial SequenceSynthetic
230Asp Ala Ser1 23127DNAArtificial SequenceSynthetic
231cagcagcgta gtgactggcc gctcact
272329PRTArtificial SequenceSynthetic 232Gln Gln Arg Ser Asp Trp Pro Leu
Thr1 5 233363DNAArtificial
SequenceSynthetic 233gaggtgcagc tgttggagtc tgggggaggc ttgctacagc
cgggggggtc cctgagactc 60tcctgtgcag cctctggaat cacctttagc acctatgcca
tgagctgggt ccgtcaggct 120ccagggaggg ggctggagtg ggtctcagct attagtggta
gtggtgatag cacatcctac 180gcagactccg tgaagggccg gttcaccagc tccagagaca
attccaagaa cacgctgtat 240ctgcaaatga acagcctgag agccgaggac acggccgtat
attactgtgc gaaagtcata 300gcagctcgtc ctcactggaa cttcgatctc tggggccgtg
gcaccctggt cactgtctcc 360tca
363234121PRTArtificial SequenceSynthetic 234Glu
Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Leu Gln Pro Gly Gly1
5 10 15 Ser Leu Arg Leu Ser Cys
Ala Ala Ser Gly Ile Thr Phe Ser Thr Tyr 20 25
30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Arg
Gly Leu Glu Trp Val 35 40 45
Ser Ala Ile Ser Gly Ser Gly Asp Ser Thr Ser Tyr Ala Asp Ser Val
50 55 60 Lys Gly Arg
Phe Thr Ser Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr65 70
75 80 Leu Gln Met Asn Ser Leu Arg Ala
Glu Asp Thr Ala Val Tyr Tyr Cys 85 90
95 Ala Lys Val Ile Ala Ala Arg Pro His Trp Asn Phe Asp
Leu Trp Gly 100 105 110
Arg Gly Thr Leu Val Thr Val Ser Ser 115 120
235324DNAArtificial SequenceSynthetic 235gaaattgtgt tgacacagtc tccagccacc
ctgtctttgt ctccagggga aagagccacc 60ctctcctgca gggccagtca gagtgttagt
agatatttag cctggtatca acagaaacct 120ggccaggctc ccaggctcct catctatgat
gcatccaaca gggccactgg catcccagcc 180aggttcagtg gcagtgggtc tgggacagac
ttcactctca ccatcagcag cctagagcct 240gaagattttg gagtttatta ctgtcagcag
cgtagtgact ggccgctcac tttcggcgga 300gggaccaagg tggagatcaa acgg
324236107PRTArtificial
SequenceSynthetic 236Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu
Ser Pro Gly1 5 10 15
Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Arg Tyr
20 25 30 Leu Ala Trp Tyr Gln
Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile 35 40
45 Tyr Asp Ala Ser Asn Arg Ala Thr Gly Ile
Pro Ala Arg Phe Ser Gly 50 55 60
Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu
Pro65 70 75 80 Glu
Asp Phe Gly Val Tyr Tyr Cys Gln Gln Arg Ser Asp Trp Pro Leu
85 90 95 Thr Phe Gly Gly Gly Thr
Lys Val Glu Ile Lys 100 105
237363DNAArtificial SequenceSynthetic 237gaggtgcagc tgttggagtc tgggggaggc
ttggtacagc cgggggggtc cctgagactc 60tcctgtgcag cctctggaat cacctttagc
acctatgcca tgagctgggt ccgtcaggct 120ccagggaagg ggctggagtg ggtctcagct
attagtggta gtggtgatag cacatactac 180gcagactccg tgaagggccg gttcaccatc
tccagagaca attccaagaa cacgctgtat 240ctgcaaatga acagcctgag agccgaggac
acggccgtat attactgtgc gaaagtcata 300gcagctcgtc ctcactggaa cttcgatctc
tggggccgtg gcaccctggt cactgtctcc 360tca
363238121PRTArtificial
SequenceSynthetic 238Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln
Pro Gly Gly1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Ile Thr Phe Ser Thr Tyr
20 25 30 Ala Met Ser Trp Val
Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40
45 Ser Ala Ile Ser Gly Ser Gly Asp Ser Thr
Tyr Tyr Ala Asp Ser Val 50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu
Tyr65 70 75 80 Leu
Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95 Ala Lys Val Ile Ala Ala
Arg Pro His Trp Asn Phe Asp Leu Trp Gly 100
105 110 Arg Gly Thr Leu Val Thr Val Ser Ser
115 120 239324DNAArtificial SequenceSynthetic
239gaaattgtgt tgacacagtc tccagccacc ctgtctttgt ctccagggga aagagccacc
60ctctcctgca gggccagtca gagtgttagt agatatttag cctggtatca acagaaacct
120ggccaggctc ccaggctcct catctatgat gcatccaaca gggccactgg catcccagcc
180aggttcagtg gcagtgggtc tgggacagac ttcactctca ccatcagcag cctagagcct
240gaagattttg cagtttatta ctgtcagcag cgtagtgact ggccgctcac tttcggcgga
300gggaccaagg tggagatcaa acgg
324240108PRTArtificial SequenceSynthetic 240Glu Ile Val Leu Thr Gln Ser
Pro Ala Thr Leu Ser Leu Ser Pro Gly1 5 10
15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser
Val Ser Arg Tyr 20 25 30
Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile
35 40 45 Tyr Asp Ala Ser
Asn Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly 50 55
60 Ser Gly Ser Gly Thr Asp Phe Thr Leu
Thr Ile Ser Ser Leu Glu Pro65 70 75
80 Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Ser Asp Trp
Pro Leu 85 90 95
Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg 100
105 241366DNAArtificial SequenceSynthetic 241caggtgcagc
tggtggagtc tgggggaggc gtggtccagc ctgggaggtc cctgagactc 60acctgtgcag
cctctggatt caccttcagt agtaatggca tgcactgggt ccgccaggct 120ccaggcaagg
ggctggagtg ggtggcaatt atatcatatg atggaaataa tcaatactat 180gcagactccg
tgaagggccg attcaccatc tccagagaca attccaagca cacgctgtat 240ctggaaatga
acagcctgag agctgaggac acggctgtgt attactgtac aaaagccatc 300tctataagtg
gaacttacaa ctggttcgat tcctggggcc agggaaccct ggtcaccgtc 360tcctca
366242122PRTArtificial SequenceSynthetic 242Gln Val Gln Leu Val Glu Ser
Gly Gly Gly Val Val Gln Pro Gly Arg1 5 10
15 Ser Leu Arg Leu Thr Cys Ala Ala Ser Gly Phe Thr
Phe Ser Ser Asn 20 25 30
Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45 Ala Ile Ile Ser
Tyr Asp Gly Asn Asn Gln Tyr Tyr Ala Asp Ser Val 50 55
60 Lys Gly Arg Phe Thr Ile Ser Arg Asp
Asn Ser Lys His Thr Leu Tyr65 70 75
80 Leu Glu Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr
Tyr Cys 85 90 95
Thr Lys Ala Ile Ser Ile Ser Gly Thr Tyr Asn Trp Phe Asp Ser Trp
100 105 110 Gly Gln Gly Thr Leu
Val Thr Val Ser Ser 115 120
24324DNAArtificial SequenceSynthetic 243ggattcacct tcagtagtaa tggc
242448PRTArtificial SequenceSynthetic
244Gly Phe Thr Phe Ser Ser Asn Gly1 5
24524DNAArtificial SequenceSynthetic 245atatcatatg atggaaataa tcaa
242468PRTArtificial SequenceSynthetic
246Ile Ser Tyr Asp Gly Asn Asn Gln1 5
24745DNAArtificial SequenceSynthetic 247acaaaagcca tctctataag tggaacttac
aactggttcg attcc 4524815PRTArtificial
SequenceSynthetic 248Thr Lys Ala Ile Ser Ile Ser Gly Thr Tyr Asn Trp Phe
Asp Ser1 5 10 15
249324DNAArtificial SequenceSynthetic 249gaaattgtat tgacacagtc tccagccatc
ctgtctttgt ctccagggga aagagccacc 60ctctcctgca gggccagtca gagtgttagc
aggtacttag cctggtacca acagaaacct 120ggccaggctc ccaggctcct catctatgat
gcatccaaca gggccactgg catcccagcc 180aggttcagtg gcagtgggtc tgggacagac
ttcactctca ccatcagcag cctagagcct 240gaagattttg cagtttatta ctgtcaacag
cgtagcaact ggccgctcac tttcggcgga 300gggaccaagg tggagatcaa acgg
324250107PRTArtificial
SequenceSynthetic 250Glu Ile Val Leu Thr Gln Ser Pro Ala Ile Leu Ser Leu
Ser Pro Gly1 5 10 15
Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Arg Tyr
20 25 30 Leu Ala Trp Tyr Gln
Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile 35 40
45 Tyr Asp Ala Ser Asn Arg Ala Thr Gly Ile
Pro Ala Arg Phe Ser Gly 50 55 60
Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu
Pro65 70 75 80 Glu
Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Ser Asn Trp Pro Leu
85 90 95 Thr Phe Gly Gly Gly Thr
Lys Val Glu Ile Lys 100 105
25118DNAArtificial SequenceSynthetic 251cagagtgtta gcaggtac
182526PRTArtificial SequenceSynthetic
252Gln Ser Val Ser Arg Tyr1 5 2539DNAArtificial
SequenceSynthetic 253gatgcatcc
92543PRTArtificial SequenceSynthetic 254Asp Ala Ser1
25527DNAArtificial SequenceSynthetic 255caacagcgta gcaactggcc
gctcact 272569PRTArtificial
SequenceSynthetic 256Gln Gln Arg Ser Asn Trp Pro Leu Thr1 5
257366DNAArtificial SequenceSynthetic 257caggtgcagc
tggtggagtc tgggggaggc gtggtccagc ctgggaggtc cctgagactc 60acctgtgcag
cctctggatt caccttcagt agtaatggca tgcactgggt ccgccaggct 120ccaggcaagg
ggctggagtg ggtggcaatt atatcatatg atggaaataa tcaatactat 180gcagactccg
tgaagggccg attcaccatc tccagagaca attccaagca cacgctgtat 240ctggaaatga
acagcctgag agctgaggac acggctgtgt attactgtac aaaagccatc 300tctataagtg
gaacttacaa ctggttcgat tcctggggcc agggaaccct ggtcaccgtc 360tcctca
366258122PRTArtificial SequenceSynthetic 258Gln Val Gln Leu Val Glu Ser
Gly Gly Gly Val Val Gln Pro Gly Arg1 5 10
15 Ser Leu Arg Leu Thr Cys Ala Ala Ser Gly Phe Thr
Phe Ser Ser Asn 20 25 30
Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45 Ala Ile Ile Ser
Tyr Asp Gly Asn Asn Gln Tyr Tyr Ala Asp Ser Val 50 55
60 Lys Gly Arg Phe Thr Ile Ser Arg Asp
Asn Ser Lys His Thr Leu Tyr65 70 75
80 Leu Glu Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr
Tyr Cys 85 90 95
Thr Lys Ala Ile Ser Ile Ser Gly Thr Tyr Asn Trp Phe Asp Ser Trp
100 105 110 Gly Gln Gly Thr Leu
Val Thr Val Ser Ser 115 120
259324DNAArtificial SequenceSynthetic 259gaaattgtat tgacacagtc tccagccatc
ctgtctttgt ctccagggga aagagccacc 60ctctcctgca gggccagtca gagtgttagc
aggtacttag cctggtacca acagaaacct 120ggccaggctc ccaggctcct catctatgat
gcatccaaca gggccactgg catcccagcc 180aggttcagtg gcagtgggtc tgggacagac
ttcactctca ccatcagcag cctagagcct 240gaagattttg cagtttatta ctgtcaacag
cgtagcaact ggccgctcac tttcggcgga 300gggaccaagg tggagatcaa acgg
324260107PRTArtificial
SequenceSynthetic 260Glu Ile Val Leu Thr Gln Ser Pro Ala Ile Leu Ser Leu
Ser Pro Gly1 5 10 15
Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Arg Tyr
20 25 30 Leu Ala Trp Tyr Gln
Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile 35 40
45 Tyr Asp Ala Ser Asn Arg Ala Thr Gly Ile
Pro Ala Arg Phe Ser Gly 50 55 60
Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu
Pro65 70 75 80 Glu
Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Ser Asn Trp Pro Leu
85 90 95 Thr Phe Gly Gly Gly Thr
Lys Val Glu Ile Lys 100 105
261366DNAArtificial SequenceSynthetic 261caggtgcagc tggtggagtc tgggggaggc
gtggtccagc ctgggaggtc cctgagactc 60tcctgtgcag cctctggatt caccttcagt
agtaatggca tgcactgggt ccgccaggct 120ccaggcaagg ggctggagtg ggtggcagtt
atatcatatg atggaaataa tcaatactat 180gcagactccg tgaagggccg attcaccatc
tccagagaca attccaagaa cacgctgtat 240ctgcaaatga acagcctgag agctgaggac
acggctgtgt attactgtac aaaagccatc 300tctataagtg gaacttacaa ctggttcgat
tcctggggcc agggaaccct ggtcaccgtc 360tcctca
366262122PRTArtificial
SequenceSynthetic 262Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln
Pro Gly Arg1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Asn
20 25 30 Gly Met His Trp Val
Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40
45 Ala Val Ile Ser Tyr Asp Gly Asn Asn Gln
Tyr Tyr Ala Asp Ser Val 50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu
Tyr65 70 75 80 Leu
Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95 Thr Lys Ala Ile Ser Ile
Ser Gly Thr Tyr Asn Trp Phe Asp Ser Trp 100
105 110 Gly Gln Gly Thr Leu Val Thr Val Ser Ser
115 120 263324DNAArtificial
SequenceSynthetic 263gaaattgtat tgacacagtc tccagccacc ctgtctttgt
ctccagggga aagagccacc 60ctctcctgca gggccagtca gagtgttagc aggtacttag
cctggtacca acagaaacct 120ggccaggctc ccaggctcct catctatgat gcatccaaca
gggccactgg catcccagcc 180aggttcagtg gcagtgggtc tgggacagac ttcactctca
ccatcagcag cctagagcct 240gaagattttg cagtttatta ctgtcaacag cgtagcaact
ggccgctcac tttcggcgga 300gggaccaagg tggagatcaa acgg
324264108PRTArtificial SequenceSynthetic 264Glu
Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly1
5 10 15 Glu Arg Ala Thr Leu Ser
Cys Arg Ala Ser Gln Ser Val Ser Arg Tyr 20 25
30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala
Pro Arg Leu Leu Ile 35 40 45
Tyr Asp Ala Ser Asn Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly
50 55 60 Ser Gly Ser
Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro65 70
75 80 Glu Asp Phe Ala Val Tyr Tyr Cys
Gln Gln Arg Ser Asn Trp Pro Leu 85 90
95 Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg
100 105 2658PRTArtificial
SequenceSynthetic 265Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa1 5
2668PRTArtificial SequenceSynthetic 266Xaa Xaa Xaa Xaa Xaa Xaa
Xaa Xaa1 5 26718PRTArtificial
SequenceSynthetic 267Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa
Xaa Xaa Xaa1 5 10 15
Xaa Xaa26811PRTArtificial SequenceSynthetic 268Xaa Xaa Xaa Xaa Xaa Xaa
Xaa Xaa Xaa Xaa Xaa1 5 10
2693PRTArtificial SequenceSynthetic 269Xaa Xaa Xaa1
2709PRTArtificial SequenceSynthetic 270Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa
Xaa1 5 271330PRTArtificial
SequenceSynthetic 271Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro
Ser Ser Lys1 5 10 15
Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr
20 25 30 Phe Pro Glu Pro Val
Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser 35 40
45 Gly Val His Thr Phe Pro Ala Val Leu Gln
Ser Ser Gly Leu Tyr Ser 50 55 60
Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln
Thr65 70 75 80 Tyr
Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys
85 90 95 Lys Val Glu Pro Lys Ser
Cys Asp Lys Thr His Thr Cys Pro Pro Cys 100
105 110 Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser
Val Phe Leu Phe Pro Pro 115 120
125 Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val
Thr Cys 130 135 140
Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp145
150 155 160 Tyr Val Asp Gly Val
Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu 165
170 175 Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val
Ser Val Leu Thr Val Leu 180 185
190 His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser
Asn 195 200 205 Lys
Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly 210
215 220 Gln Pro Arg Glu Pro Gln
Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu225 230
235 240 Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu
Val Lys Gly Phe Tyr 245 250
255 Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn
260 265 270 Asn Tyr Lys
Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe 275
280 285 Leu Tyr Ser Lys Leu Thr Val Asp
Lys Ser Arg Trp Gln Gln Gly Asn 290 295
300 Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn
His Tyr Thr305 310 315
320 Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 325
330 272327PRTArtificial SequenceSynthetic 272Ala Ser Thr Lys Gly Pro
Ser Val Phe Pro Leu Ala Pro Cys Ser Arg1 5
10 15 Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys
Leu Val Lys Asp Tyr 20 25 30
Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser
35 40 45 Gly Val His
Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser 50
55 60 Leu Ser Ser Val Val Thr Val Pro
Ser Ser Ser Leu Gly Thr Lys Thr65 70 75
80 Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys
Val Asp Lys 85 90 95
Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro Ser Cys Pro Ala Pro
100 105 110 Glu Phe Leu Gly Gly
Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 115
120 125 Asp Thr Leu Met Ile Ser Arg Thr Pro
Glu Val Thr Cys Val Val Val 130 135
140 Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp
Tyr Val Asp145 150 155
160 Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe
165 170 175 Asn Ser Thr Tyr
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 180
185 190 Trp Leu Asn Gly Lys Glu Tyr Lys Cys
Lys Val Ser Asn Lys Gly Leu 195 200
205 Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln
Pro Arg 210 215 220
Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys225
230 235 240 Asn Gln Val Ser Leu
Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp 245
250 255 Ile Ala Val Glu Trp Glu Ser Asn Gly Gln
Pro Glu Asn Asn Tyr Lys 260 265
270 Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr
Ser 275 280 285 Arg
Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser 290
295 300 Cys Ser Val Met His Glu
Ala Leu His Asn His Tyr Thr Gln Lys Ser305 310
315 320 Leu Ser Leu Ser Leu Gly Lys
325 273327PRTArtificial SequenceSynthetic 273Ala Ser Thr Lys Gly
Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg1 5
10 15 Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly
Cys Leu Val Lys Asp Tyr 20 25
30 Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr
Ser 35 40 45 Gly
Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser 50
55 60 Leu Ser Ser Val Val Thr
Val Pro Ser Ser Ser Leu Gly Thr Lys Thr65 70
75 80 Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn
Thr Lys Val Asp Lys 85 90
95 Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro
100 105 110 Glu Phe Leu
Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 115
120 125 Asp Thr Leu Met Ile Ser Arg Thr
Pro Glu Val Thr Cys Val Val Val 130 135
140 Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp
Tyr Val Asp145 150 155
160 Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe
165 170 175 Asn Ser Thr Tyr
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 180
185 190 Trp Leu Asn Gly Lys Glu Tyr Lys Cys
Lys Val Ser Asn Lys Gly Leu 195 200
205 Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln
Pro Arg 210 215 220
Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys225
230 235 240 Asn Gln Val Ser Leu
Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp 245
250 255 Ile Ala Val Glu Trp Glu Ser Asn Gly Gln
Pro Glu Asn Asn Tyr Lys 260 265
270 Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr
Ser 275 280 285 Arg
Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser 290
295 300 Cys Ser Val Met His Glu
Ala Leu His Asn His Tyr Thr Gln Lys Ser305 310
315 320 Leu Ser Leu Ser Leu Gly Lys
325 274207PRTHomo sapiens 274Met Lys Val Leu Gln Glu Pro Thr Cys
Val Ser Asp Tyr Met Ser Ile1 5 10
15 Ser Thr Cys Glu Trp Lys Met Asn Gly Pro Thr Asn Cys Ser
Thr Glu 20 25 30
Leu Arg Leu Leu Tyr Gln Leu Val Phe Leu Leu Ser Glu Ala His Thr 35
40 45 Cys Ile Pro Glu Asn
Asn Gly Gly Ala Gly Cys Val Cys His Leu Leu 50 55
60 Met Asp Asp Val Val Ser Ala Asp Asn Tyr
Thr Leu Asp Leu Trp Ala65 70 75
80 Gly Gln Gln Leu Leu Trp Lys Gly Ser Phe Lys Pro Ser Glu His
Val 85 90 95 Lys
Pro Arg Ala Pro Gly Asn Leu Thr Val His Thr Asn Val Ser Asp
100 105 110 Thr Leu Leu Leu Thr
Trp Ser Asn Pro Tyr Pro Pro Asp Asn Tyr Leu 115
120 125 Tyr Asn His Leu Thr Tyr Ala Val Asn
Ile Trp Ser Glu Asn Asp Pro 130 135
140 Ala Asp Phe Arg Ile Tyr Asn Val Thr Tyr Leu Glu Pro
Ser Leu Arg145 150 155
160 Ile Ala Ala Ser Thr Leu Lys Ser Gly Ile Ser Tyr Arg Ala Arg Val
165 170 175 Arg Ala Trp Ala
Gln Cys Tyr Asn Thr Thr Trp Ser Glu Trp Ser Pro 180
185 190 Ser Thr Lys Trp His Asn Ser Tyr Arg
Glu Pro Phe Glu Gln His 195 200
205 275231PRTMacaca fasicularis 275Met Gly Trp Leu Cys Ser Gly
Leu Leu Phe Pro Val Ser Cys Leu Val1 5 10
15 Leu Leu Gln Val Ala Ser Ser Gly Ser Met Lys Val
Leu Gln Glu Pro 20 25 30
Thr Cys Val Ser Asp Tyr Met Ser Ile Ser Thr Cys Glu Trp Lys Met
35 40 45 Gly Gly Pro Thr
Asn Cys Ser Ala Glu Leu Arg Leu Leu Tyr Gln Leu 50 55
60 Val Phe Gln Ser Ser Glu Thr His Thr
Cys Val Pro Glu Asn Asn Gly65 70 75
80 Gly Val Gly Cys Val Cys His Leu Leu Met Asp Asp Val Val
Ser Met 85 90 95
Asp Asn Tyr Thr Leu Asp Leu Trp Ala Gly Gln Gln Leu Leu Trp Lys
100 105 110 Gly Ser Phe Lys Pro
Ser Glu His Val Lys Pro Arg Ala Pro Gly Asn 115
120 125 Leu Thr Val His Thr Asn Val Ser Asp
Thr Val Leu Leu Thr Trp Ser 130 135
140 Asn Pro Tyr Pro Pro Asp Asn Tyr Leu Tyr Asn Asp Leu
Thr Tyr Ala145 150 155
160 Val Asn Ile Trp Ser Glu Asn Asp Pro Ala Tyr Ser Arg Ile His Asn
165 170 175 Val Thr Tyr Leu
Lys Pro Thr Leu Arg Ile Pro Ala Ser Thr Leu Lys 180
185 190 Ser Gly Ile Ser Tyr Arg Ala Arg Val
Arg Ala Trp Ala Gln His Tyr 195 200
205 Asn Thr Thr Trp Ser Glu Trp Ser Pro Ser Thr Lys Trp Tyr
Asn Ser 210 215 220
Tyr Arg Glu Pro Phe Glu Gln225 230
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