Patent Publication Number: US-2002009758-A1

Title: Compositions and methods for the therapy and diagnosis of lung cancer

Description:
CROSS REFERENCE TO RELATED APPLICATIONS  
     [0001] This application is related to U.S. Provisional Application No. 60/207,485, filed May 26, 2000 and U.S. Provisional Application No. 60/230,475, filed Sep. 6, 2000, incorporated in their entirety herein by reference. 
    
    
     
       TECHNICAL FIELD OF THE INVENTION  
       [0002] The present invention relates generally to therapy and diagnosis of cancer, such as lung cancer. The invention is more specifically related to polypeptides, comprising at least a portion of a lung tumor protein, and to polynucleotides encoding such polypeptides. Such polypeptides and polynucleotides are useful in pharmaceutical compositions, e.g., vaccines, and other compositions for the diagnosis and treatment of lung cancer.  
       BACKGROUND OF THE INVENTION  
       [0003] Cancer is a significant health problem throughout the world. Although advances have been made in detection and therapy of cancer, no vaccine or other universally successful method for prevention and/or treatment is currently available. Current therapies, which are generally based on a combination of chemotherapy or surgery and radiation, continue to prove inadequate in many patients.  
       [0004] Lung cancer is a significant health problem throughout the world. In the U.S., lung cancer is the primary cause of cancer death among both men and women, with an estimated 172,000 new cases being reported in 1994. The five-year survival rate among all lung cancer patients, regardless of the stage of disease at diagnosis, is only 13%. This contrasts with a five-year survival rate of 46% among cases detected while the disease is still localized. However, early detection of lung cancer is difficult since clinical symptoms are often not seen until the disease has reached an advanced stage, and only 16% of lung cancers are discovered before the disease has spread.  
       [0005] In spite of considerable research into therapies for these and other cancers, lung cancer remains difficult to diagnose and treat effectively. Accordingly, there is a need in the art for improved methods for detecting and treating such cancers. The present invention fulfills these needs and further provides other related advantages.  
       SUMMARY OF THE INVENTION  
       [0006] In one aspect, the present invention provides polynucleotide compositions comprising a sequence selected from the group consisting of:  
       [0007] (a) sequences provided in SEQ ID NO:1-35, 42-55, 58-60, 63-91 and 93-95;  
       [0008] (b) complements of the sequences provided in SEQ ID NO:1-35, 42-55, 58-60, 63-91 and 93-95;  
       [0009] (c) sequences consisting of at least 20, 25, 30, 35, 40, 45, 50, 75 and 100 contiguous residues of a sequence provided in SEQ ID NO: 1-35, 42-55, 58-60, 63-91 and 93-95;  
       [0010] (d) sequences that hybridize to a sequence provided in SEQ ID NO: 1-35, 42-55, 58-60, 63-91 and 93-95, under moderate or highly stringent conditions;  
       [0011] (e) sequences having at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identity to a sequence of SEQ ID NO:1-35, 42-55, 58-60, 63-91 and 93-95; and  
       [0012] (f) degenerate variants of a sequence provided in SEQ ID NO: 1-35, 42-55, 58-60, 63-91 and 93-95.  
       [0013] In one preferred embodiment, the polynucleotide compositions of the invention are expressed in at least about 20%, more preferably in at least about 30%, and most preferably in at least about 50% of lung tumors samples tested, at a level that is at least about 2-fold, preferably at least about 5-fold, and most preferably at least about 10-fold higher than that for normal tissues.  
       [0014] The present invention, in another aspect, provides polypeptide compositions comprising an amino acid sequence that is encoded by a polynucleotide sequence described above.  
       [0015] The present invention further provides polypeptide compositions comprising an amino acid sequence selected from the group consisting of sequences recited in SEQ ID NO:36-41, 56, 57, 61, 62, 92 and 96.  
       [0016] In certain preferred embodiments, the polypeptides and/or polynucleotides of the present invention are immunogenic, i.e., they are capable of eliciting an immune response, particularly a humoral and/or cellular immune response, as further described herein.  
       [0017] The present invention further provides fragments, variants and/or derivatives of the disclosed polypeptide and/or polynucleotide sequences, wherein the fragments, variants and/or derivatives preferably have a level of immunogenic activity of at least about 50%, preferably at least about 70% and more preferably at least about 90% of the level of immunogenic activity of a polypeptide sequence set forth in SEQ ID NO:36-41, 56, 57, 61, 62, 92 and 96 or a polypeptide sequence encoded by a polynucleotide sequence set forth in SEQ ID NO:1-35, 42-55, 58-60, 63-91 and 93-95.  
       [0018] The present invention further provides polynucleotides that encode a polypeptide described above, expression vectors comprising such polynucleotides and host cells transformed or transfected with such expression vectors.  
       [0019] Within other aspects, the present invention provides pharmaceutical compositions comprising a polypeptide or polynucleotide as described above and a physiologically acceptable carrier.  
       [0020] Within a related aspect of the present invention, the pharmaceutical compositions, e.g., vaccine compositions, are provided for prophylactic or therapeutic applications. Such compositions generally comprise an immunogenic polypeptide or polynucleotide of the invention and an immunostimulant, such as an adjuvant.  
       [0021] The present invention further provides pharmaceutical compositions that comprise: (a) an antibody or antigen-binding fragment thereof that specifically binds to a polypeptide of the present invention, or a fragment thereof; and (b) a physiologically acceptable carrier.  
       [0022] Within further aspects, the present invention provides pharmaceutical compositions comprising: (a) an antigen presenting cell that expresses a polypeptide as described above and (b) a pharmaceutically acceptable carrier or excipient. Illustrative antigen presenting cells include dendritic cells, macrophages, monocytes, fibroblasts and B cells.  
       [0023] Within related aspects, pharmaceutical compositions are provided that comprise: (a) an antigen presenting cell that expresses a polypeptide as described above and (b) an immunostimulant.  
       [0024] The present invention further provides, in other aspects, fusion proteins that comprise at least one polypeptide as described above, as well as polynucleotides encoding such fusion proteins, typically in the form of pharmaceutical compositions, e.g., vaccine compositions, comprising a physiologically acceptable carrier and/or an immunostimulant. The fusions proteins may comprise multiple immunogenic polypeptides or portions/variants thereof, as described herein, and may further comprise one or more polypeptide segments for facilitating the expression, purification and/or immunogenicity of the polypeptide(s).  
       [0025] Within further aspects, the present invention provides methods for stimulating an immune response in a patient, preferably a T cell response in a human patient, comprising administering a pharmaceutical composition described herein. The patient may be afflicted with lung cancer, in which case the methods provide treatment for the disease, or patient considered at risk for such a disease may be treated prophylactically.  
       [0026] Within further aspects, the present invention provides methods for inhibiting the development of a cancer in a patient, comprising administering to a patient a pharmaceutical composition as recited above. The patient may be afflicted with lung cancer, in which case the methods provide treatment for the disease, or patient considered at risk for such a disease may be treated prophylactically.  
       [0027] The present invention further provides, within other aspects, methods for removing tumor cells from a biological sample, comprising contacting a biological sample with T cells that specifically react with a polypeptide of the present invention, wherein the step of contacting is performed under conditions and for a time sufficient to permit the removal of cells expressing the protein from the sample.  
       [0028] Within related aspects, methods are provided for inhibiting the development of a cancer in a patient, comprising administering to a patient a biological sample treated as described above.  
       [0029] Methods are further provided, within other aspects, for stimulating and/or expanding T cells specific for a polypeptide of the present invention, comprising contacting T cells with one or more of: (i) a polypeptide as described above; (ii) a polynucleotide encoding such a polypeptide; and/or (iii) an antigen presenting cell that expresses such a polypeptide; under conditions and for a time sufficient to permit the stimulation and/or expansion of T cells. Isolated T cell populations comprising T cells prepared as described above are also provided.  
       [0030] Within further aspects, the present invention provides methods for inhibiting the development of a cancer in a patient, comprising administering to a patient an effective amount of a T cell population as described above.  
       [0031] The present invention further provides methods for inhibiting the development of a cancer in a patient, comprising the steps of: (a) incubating CD4 +  and/or CD8 +  T cells isolated from a patient with one or more of: (i) a polypeptide comprising at least an immunogenic portion of polypeptide disclosed herein; (ii) a polynucleotide encoding such a polypeptide; and (iii) an antigen-presenting cell that expressed such a polypeptide; and (b) administering to the patient an effective amount of the proliferated T cells, and thereby inhibiting the development of a cancer in the patient. Proliferated cells may, but need not, be cloned prior to administration to the patient.  
       [0032] Within further aspects, the present invention provides methods for determining the presence or absence of a cancer, preferably a lung cancer, in a patient comprising: (a) contacting a biological sample obtained from a patient with a binding agent that binds to a polypeptide as recited above; (b) detecting in the sample an amount of polypeptide that binds to the binding agent; and (c) comparing the amount of polypeptide with a predetermined cut-off value, and therefrom determining the presence or absence of a cancer in the patient. Within preferred embodiments, the binding agent is an antibody, more preferably a monoclonal antibody.  
       [0033] The present invention also provides, within other aspects, methods for monitoring the progression of a cancer in a patient. Such methods comprise the steps of: (a) contacting a biological sample obtained from a patient at a first point in time with a binding agent that binds to a polypeptide as recited above; (b) detecting in the sample an amount of polypeptide that binds to the binding agent; (c) repeating steps (a) and (b) using a biological sample obtained from the patient at a subsequent point in time; and (d) comparing the amount of polypeptide detected in step (c) with the amount detected in step (b) and therefrom monitoring the progression of the cancer in the patient.  
       [0034] The present invention further provides, within other aspects, methods for determining the presence or absence of a cancer in a patient, comprising the steps of: (a) contacting a biological sample, e.g., tumor sample, serum sample, etc., obtained from a patient with an oligonucleotide that hybridizes to a polynucleotide that encodes a polypeptide of the present invention; (b) detecting in the sample a level of a polynucleotide, preferably mRNA, that hybridizes to the oligonucleotide; and (c) comparing the level of polynucleotide that hybridizes to the oligonucleotide with a predetermined cut-off value, and therefrom determining the presence or absence of a cancer in the patient. Within certain embodiments, the amount of mRNA is detected via polymerase chain reaction using, for example, at least one oligonucleotide primer that hybridizes to a polynucleotide encoding a polypeptide as recited above, or a complement of such a polynucleotide. Within other embodiments, the amount of mRNA is detected using a hybridization technique, employing an oligonucleotide probe that hybridizes to a polynucleotide that encodes a polypeptide as recited above, or a complement of such a polynucleotide.  
       [0035] In related aspects, methods are provided for monitoring the progression of a cancer in a patient, comprising the steps of: (a) contacting a biological sample obtained from a patient with an oligonucleotide that hybridizes to a polynucleotide that encodes a polypeptide of the present invention; (b) detecting in the sample an amount of a polynucleotide that hybridizes to the oligonucleotide; (c) repeating steps (a) and (b) using a biological sample obtained from the patient at a subsequent point in time; and (d) comparing the amount of polynucleotide detected in step (c) with the amount detected in step (b) and therefrom monitoring the progression of the cancer in the patient.  
       [0036] Within further aspects, the present invention provides antibodies, such as monoclonal antibodies, that bind to a polypeptide as described above, as well as diagnostic kits comprising such antibodies. Diagnostic kits comprising one or more oligonucleotide probes or primers as described above are also provided.  
       [0037] These and other aspects of the present invention will become apparent upon reference to the following detailed description and attached drawings. All references disclosed herein are hereby incorporated by reference in their entirety as if each was incorporated individually.  
       SEQUENCE IDENTIFIERS  
       [0038] SEQ ID NO: 1 is the cDNA sequence for Clone ID # 55964 which is named clone L1040C, and is the same sequence as SEQ ID NO:2337 from U.S. Provisional Application No. 60/207,485.  
       [0039] SEQ ID NO:2 is an extended cDNA sequence for L1040C (Clone ID # 55964).  
       [0040] SEQ ID NO:3 is the cDNA sequence for Clone ID # 58269 which is named clone L1039C, and is the same sequence as SEQ ID NO:7264 from U.S. Provisional Application No. 60/207,485.  
       [0041] SEQ ID NO:4 is an extended cDNA sequence for L1039C (Clone ID # 58269), and which corresponds to the fbx5 F-box gene.  
       [0042] SEQ ID NO:5 is the cDNA sequence for Clone ID # 58267 which is named clone L1037C, and is the same sequence as SEQ ID NO:4978 from U.S. Provisional Application No. 60/207,485.  
       [0043] SEQ ID NO:6 is an extended cDNA sequence for L1037C (Clone # 58267), and which corresponds to the mitotic checkpoint kinase mad3-like gene.  
       [0044] SEQ ID NO:7 is the cDNA sequence for Clone ID # 58245 which is named clone L1038C, and is the same sequence as SEQ ID NO:1796 from U.S. Provisional Application No. 60/207,485.  
       [0045] SEQ ID NO:8 is an extended cDNA sequence for L1038C (Clone ID # 58245), and which corresponds to a neuronal ER localized gene.  
       [0046] SEQ ID NO:9 is the cDNA sequence for Clone ID # 55571 which is named clone L1027C, and is the same sequence as SEQ ID NO:4538 from U.S. Provisional Application No. 60/207,485.  
       [0047] SEQ ID NO:10 is an extended cDNA sequence for L1027C (Clone ID # 55571).  
       [0048] SEQ ID NO: 11 is the cDNA sequence for Clone ID # 55978.  
       [0049] SEQ ID NO:12 is an extended cDNA sequence for Clone ID # 55978.  
       [0050] SEQ ID NO:13 is the cDNA sequence for Clone ID # 55980.  
       [0051] SEQ ID NO:14 is an extended cDNA sequence for Clone ID # 55980.  
       [0052] SEQ ID NO:15 is the cDNA sequence for Clone ID # 58346.  
       [0053] SEQ ID NO:16 is an extended cDNA sequence for Clone ID # 58346.  
       [0054] SEQ ID NO:17 is the cDNA sequence for Clone ID # 55561.  
       [0055] SEQ ID NO: 18 is an extended cDNA sequence for Clone ID # 55561.  
       [0056] SEQ ID NO:19 is the cDNA sequence for Clone ID # 55984.  
       [0057] SEQ ID NO:20 is an extended cDNA sequence for Clone ID # 55984, and which corresponds to a gt mismatch glycosylase gene.  
       [0058] SEQ ID NO:21 is the cDNA sequence for Clone ID # 58261.  
       [0059] SEQ ID NO:22 is an extended cDNA sequence for Clone ID # 58261, and which corresponds to a phosphoserine aminotransferase gene.  
       [0060] SEQ ID NO:23 is the cDNA sequence for Clone ID # 58348.  
       [0061] SEQ ID NO:24 is an extended cDNA sequence for Clone ID # 58348, and which corresponds to a hCAP gene.  
       [0062] SEQ ID NO:25 is the cDNA sequence for Clone ID # 56016.  
       [0063] SEQ ID NO:26 is an extended cDNA sequence for Clone ID # 56016.  
       [0064] SEQ ID NO:27 is the cDNA sequence for Clone ID # 55987.  
       [0065] SEQ ID NO:28 is an extended cDNA sequence for Clone ID # 55987.  
       [0066] SEQ ID NO:29 is the cDNA sequence for Clone ID # 55956.  
       [0067] SEQ ID NO:30 is an extended cDNA sequence for Clone ID # 55956.  
       [0068] SEQ ID NO:31 is the cDNA sequence for Clone ID # 55952.  
       [0069] SEQ ID NO:32 is the cDNA sequence for Clone ID # 55957.  
       [0070] SEQ ID NO:33 is an extended cDNA sequence for Clone ID # 55957.  
       [0071] SEQ ID NO:34 is the cDNA sequence for Clone ID # 55559.  
       [0072] SEQ ID NO:35 is an extended cDNA sequence for Clone ID # 55559.  
       [0073] SEQ ID NO:36 is an amino acid sequence of an ORF for L1027C, encoded by the polynucleotide of SEQ ID NO: 10.  
       [0074] SEQ ID NO:37 is an amino acid sequence of the F-box protein Fbx5 encoded by SEQ ID NO:4.  
       [0075] SEQ ID NO:38 is an amino acid sequence of the mitotic checkpoint kinase MAD3-like protein encoded by SEQ ID NO:6.  
       [0076] SEQ ID NO:39 is an amino acid sequence of the neuronal olfactomedin-related ER localized protein encoded by SEQ ID NO:8.  
       [0077] SEQ ID NO:40 is an amino acid sequence of the phosphoserine aminotransferase encoded by SEQ ID NO:22.  
       [0078] SEQ ID NO:41 is an amino acid sequence of the gt mismatch glycosylase encoded by SEQ ID NO:20.  
       [0079] SEQ ID NO:42 is the determined cDNA sequence for Clone ID # 63575 which is named clone L1053 C.  
       [0080] SEQ ID NO:43 is the determined cDNA sequence for Clone ID # 63582 which is named clone L1054C.  
       [0081] SEQ ID NO:44 is the determined cDNA sequence for Clone ID # 63598 which is named clone L1055C.  
       [0082] SEQ ID NO:45 is the determined cDNA sequence for Clone ID # 64963 which is named clone L1056C.  
       [0083] SEQ ID NO:46 is the determined cDNA sequence for Clone ID # 64988 which is named clone L1058C.  
       [0084] SEQ ID NO:47 is the determined cDNA sequence for Clone ID # 63485.  
       [0085] SEQ ID NO:48 is the determined cDNA sequence for Clone ID # 65010.  
       [0086] SEQ ID NO:49 is a predicted full-length cDNA sequence for SEQ ID NO:42 which is a full-length sequence from Genbank for an insulinoma-associated 1 mRNA.  
       [0087] SEQ ID NO:50 is a predicted full-length cDNA sequence for SEQ ID NO:43 which is a full-length sequence from Genbank for KIAA0535.  
       [0088] SEQ ID NO:51 is a predicted extended cDNA sequence for SEQ ID NO:44.  
       [0089] SEQ ID NO:52 is a a predicted full-length cDNA sequence for SEQ ID NO:45 which is a full-length sequence from genbank for a human DAZ mRNA 3′UTR.  
       [0090] SEQ ID NO:53 is a predicted extended cDNA sequence for SEQ ID NO:46.  
       [0091] SEQ ID NO:54 is a predicted extended cDNA sequence for SEQ ID NO:47.  
       [0092] SEQ ID NO:55 is a predicted extended cDNA sequence for SEQ ID NO:48.  
       [0093] SEQ ID NO:56 is the deduced amino acid sequence encoded by SEQ ID NO:49.  
       [0094] SEQ ID NO:57 is the deduced amino acid sequence encoded by SEQ ID NO:50.  
       [0095] SEQ ID NO:58 is the determined full-length cDNA sequence for clone L1058C (sequence of the originally isolated clone is given in SEQ ID NO:46 and the predicted extended cDNA sequence in SEQ ID NO:53).  
       [0096] SEQ ID NO:59 is a first predicted ORF of SEQ ID NO:58.  
       [0097] SEQ ID NO:60 is a second predicted ORF of SEQ ID NO:58.  
       [0098] SEQ ID NO:61 is the deduced amino acid sequence encoded by SEQ ID NO:59.  
       [0099] SEQ ID NO:62 is the deduced amino acid sequence encoded by SEQ ID NO:60.  
       [0100] SEQ ID NO:63 is the determined cDNA sequence for Clone ID # 72761.  
       [0101] SEQ ID NO:64 is the determined cDNA sequence for Clone ID # 72762.  
       [0102] SEQ ID NO:65 is the determined cDNA sequence for Clone ID # 72763.  
       [0103] SEQ ID NO:66 is the determined cDNA sequence for Clone ID # 72764.  
       [0104] SEQ ID NO:67 is the determined cDNA sequence for Clone ID # 72765.  
       [0105] SEQ ID NO:68 is the determined cDNA sequence for Clone ID # 72766.  
       [0106] SEQ ID NO:69 is the determined cDNA sequence for Clone ID # 72772.  
       [0107] SEQ ID NO:70 is the determined cDNA sequence for Clone ID # 72775.  
       [0108] SEQ ID NO:71 is the determined cDNA sequence for Clone ID # 72776.  
       [0109] SEQ ID NO:72 is the determined cDNA sequence for Clone ID # 72779.  
       [0110] SEQ ID NO:73 is the determined cDNA sequence for Clone ID # 72781.  
       [0111] SEQ ID NO:74 is the determined cDNA sequence for Clone ID # 72784.  
       [0112] SEQ ID NO:75 is the determined cDNA sequence for Clone ID # 72788.  
       [0113] SEQ ID NO:76 is the determined cDNA sequence for Clone ID # 72789.  
       [0114] SEQ ID NO:77 is the determined cDNA sequence for Clone ID 72790.  
       [0115] SEQ ID NO:78 is the determined cDNA sequence for Clone ID # 72791.  
       [0116] SEQ ID NO:79 is the determined cDNA sequence for Clone ID # 72792.  
       [0117] SEQ ID NO:80 is the determined cDNA sequence for Clone ID 72794.  
       [0118] SEQ ID NO:81 is the determined cDNA sequence for Clone ID # 72795.  
       [0119] SEQ ID NO: 82 is the determined cDNA sequence for Clone ID #72797.  
       [0120] SEQ ID NO:83 is the determined cDNA sequence for Clone ID # 72798.  
       [0121] SEQ ID NO:84 is the determined cDNA sequence for Clone ID # 72804.  
       [0122] SEQ ID NO:85 is the determined cDNA sequence for Clone ID # 72805.  
       [0123] SEQ ID NO:86 is the determined cDNA sequence for Clone ID # 72806.  
       [0124] SEQ ID NO:87 is the determined cDNA sequence for Clone ID # 72807.  
       [0125] SEQ ID NO:88 is the determined CDNA sequence for Clone ID # 72808.  
       [0126] SEQ ID NO:89 is the determined cDNA sequence for Clone ID # 72809.  
       [0127] SEQ ID NO:90 is the determined cDNA sequence for Clone ID # 72811.  
       [0128] SEQ ID NO:91 is the determined full-length cDNA sequence for Clone ID 72813 which is named clone L1080C.  
       [0129] SEQ ID NO:92 is the deduced amino acid sequence encoded by SEQ ID NO:91.  
       [0130] SEQ ID NO:93 is the ORF for L1027C.  
       [0131] SEQ ID NO:94 is a first determined full-length cDNA sequence for L1027C.  
       [0132] SEQ ID NO:95 is a second determined full-length cDNA sequence for L1027C.  
       [0133] SEQ ID NO:96 is the deduced amino acid sequence encoded by SEQ ID NO:93.  
       DETAILED DESCRIPTION OF THE INVENTION  
       [0134] The present invention is directed generally to compositions and their use in the therapy and diagnosis of cancer, particularly lung cancer. As described further below, illustrative compositions of the present invention include, but are not restricted to, polypeptides, particularly immunogenic polypeptides, polynucleotides encoding such polypeptides, antibodies and other binding agents, antigen presenting cells (APCs) and immune system cells (e.g., T cells).  
       [0135] The practice of the present invention will employ, unless indicated specifically to the contrary, conventional methods of virology, immunology, microbiology, molecular biology and recombinant DNA techniques within the skill of the art, many of which are described below for the purpose of illustration. Such techniques are explained fully in the literature. See, e.g., Sambrook, et al. Molecular Cloning: A Laboratory Manual (2nd Edition, 1989); Maniatis et al. Molecular Cloning: A Laboratory Manual (1982); DNA Cloning: A Practical Approach, vol. I &amp; II (D. Glover, ed.); Oligonucleotide Synthesis (N. Gait, ed., 1984); Nucleic Acid Hybridization (B. Hames &amp; S. Higgins, eds., 1985); Transcription and Translation (B. Hames &amp; S. Higgins, eds., 1984); Animal Cell Culture (R. Freshney, ed., 1986); Perbal, A Practical Guide to Molecular Cloning (1984).  
       [0136] All publications, patents and patent applications cited herein, whether supra or infra, are hereby incorporated by reference in their entirety.  
       [0137] As used in this specification and the appended claims, the singular forms “a,” “an” and “the” include plural references unless the content clearly dictates otherwise.  
       [0138] Polypeptide Compositions  
       [0139] As used herein, the term “polypeptide” “is used in its conventional meaning, i.e., as a sequence of amino acids. The polypeptides are not limited to a specific length of the product; thus, peptides, oligopeptides, and proteins are included within the definition of polypeptide, and such terms may be used interchangeably herein unless specifically indicated otherwise. This term also does not refer to or exclude post-expression modifications of the polypeptide, for example, glycosylations, acetylations, phosphorylations and the like, as well as other modifications known in the art, both naturally occurring and non-naturally occurring. A polypeptide may be an entire protein, or a subsequence thereof. Particular polypeptides of interest in the context of this invention are amino acid subsequences comprising epitopes, i.e., antigenic determinants substantially responsible for the immunogenic properties of a polypeptide and being capable of evoking an immune response.  
       [0140] Particularly illustrative polypeptides of the present invention comprise those encoded by a polynucleotide sequence set forth in any one of SEQ ID NO:1-35, 42-55, 58-60, 63-91 and 93-95, or a sequence that hybridizes under moderately stringent conditions, or, alternatively, under highly stringent conditions, to a polynucleotide sequence set forth in any one of SEQ ID NO:1-35, 42-55, 58-60, 63-91 and 93-95. Certain other illustrative polypeptides of the invention comprise amino acid sequences as set forth in any one of SEQ ID NOs:36-41, 56, 57, 61, 62, 92 and 96.  
       [0141] The polypeptides of the present invention are sometimes herein referred to as lung tumor proteins or lung tumor polypeptides, as an indication that their identification has been based at least in part upon their increased levels of expression in lung tumor samples. Thus, a “lung tumor polypeptide” or “lung tumor protein,” refers generally to a polypeptide sequence of the present invention, or a polynucleotide sequence encoding such a polypeptide, that is expressed in a substantial proportion of lung tumor samples, for example preferably greater than about 20%, more preferably greater than about 30%, and most preferably greater than about 50% or more of lung tumor samples tested, at a level that is at least two fold, and preferably at least five fold, greater than the level of expression in normal tissues, as determined using a representative assay provided herein. A lung tumor polypeptide sequence of the invention, based upon its increased level of expression in tumor cells, has particular utility both as a diagnostic marker as well as a therapeutic target, as further described below.  
       [0142] In certain preferred embodiments, the polypeptides of the invention are immunogenic, i.e., they react detectably within an immunoassay (such as an ELISA or T-cell stimulation assay) with antisera and/or T-cells from a patient with lung cancer. Screening for immunogenic activity can be performed using techniques well known to the skilled artisan. For example, such screens can be performed using methods such as those described in Harlow and Lane,  Antibodies: A Laboratory Manual,  Cold Spring Harbor Laboratory, 1988. In one illustrative example, a polypeptide may be immobilized on a solid support and contacted with patient sera to allow binding of antibodies within the sera to the immobilized polypeptide. Unbound sera may then be removed and bound antibodies detected using, for example,  125 I-labeled Protein A.  
       [0143] As would be recognized by the skilled artisan, immunogenic portions of the polypeptides disclosed herein are also encompassed by the present invention. An “immunogenic portion,” as used herein, is a fragment of an immunogenic polypeptide of the invention that itself is immunologically reactive (i.e., specifically binds) with the B-cells and/or T-cell surface antigen receptors that recognize the polypeptide. Immunogenic portions may generally be identified using well known techniques, such as those summarized in Paul,  Fundamental Immunology,  3rd ed., 243-247 (Raven Press, 1993) and references cited therein. Such techniques include screening polypeptides for the ability to react with antigen-specific antibodies, antisera and/or T-cell lines or clones. As used herein, antisera and antibodies are “antigen-specific” if they specifically bind to an antigen (i.e., they react with the protein in an ELISA or other immunoassay, and do not react detectably with unrelated proteins). Such antisera and antibodies may be prepared as described herein, and using well-known techniques.  
       [0144] In one preferred embodiment, an immunogenic portion of a polypeptide of the present invention is a portion that reacts with antisera and/or T-cells at a level that is not substantially less than the reactivity of the full-length polypeptide (e.g., in an ELISA and/or T-cell reactivity assay). Preferably, the level of immunogenic activity of the immunogenic portion is at least about 50%, preferably at least about 70% and most preferably greater than about 90% of the immunogenicity for the full-length polypeptide. In some instances, preferred immunogenic portions will be identified that have a level of immunogenic activity greater than that of the corresponding full-length polypeptide, e.g., having greater than about 100% or 150% or more immunogenic activity.  
       [0145] In certain other embodiments, illustrative immunogenic portions may include peptides in which an N-terminal leader sequence and/or transmembrane domain have been deleted. Other illustrative immunogenic portions will contain a small N- and/or C-terminal deletion (e.g., 1-30 amino acids, preferably 5-15 amino acids), relative to the mature protein.  
       [0146] In another embodiment, a polypeptide composition of the invention may also comprise one or more polypeptides that are immunologically reactive with T cells and/or antibodies generated against a polypeptide of the invention, particularly a polypeptide having an amino acid sequence disclosed herein, or to an immunogenic fragment or variant thereof.  
       [0147] In another embodiment of the invention, polypeptides are provided that comprise one or more polypeptides that are capable of eliciting T cells and/or antibodies that are immunologically reactive with one or more polypeptides described herein, or one or more polypeptides encoded by contiguous nucleic acid sequences contained in the polynucleotide sequences disclosed herein, or immunogenic fragments or variants thereof, or to one or more nucleic acid sequences which hybridize to one or more of these sequences under conditions of moderate to high stringency.  
       [0148] The present invention, in another aspect, provides polypeptide fragments comprising at least about 5, 10, 15, 20, 25, 50, or 100 contiguous amino acids, or more, including all intermediate lengths, of a polypeptide compositions set forth herein, such as those set forth in SEQ ID NOs:36-41, 56, 57, 61, 62, 92 and 96, or those encoded by a polynucleotide sequence set forth in a sequence of SEQ ID NOs:1-35, 42-55, 58-60, 63-91 and 93-95.  
       [0149] In another aspect, the present invention provides variants of the polypeptide compositions described herein. Polypeptide variants generally encompassed by the present invention will typically exhibit at least about 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% or more identity (determined as described below), along its length, to a polypeptide sequences set forth herein.  
       [0150] In one preferred embodiment, the polypeptide fragments and variants provided by the present invention are immunologically reactive with an antibody and/or T-cell that react with a full-length polypeptide specifically set forth herein.  
       [0151] In another preferred embodiment, the polypeptide fragments and variants provided by the present invention exhibit a level of immunogenic activity of at least about 50%, preferably at least about 70%, and most preferably at least about 90% or more of that exhibited by a full-length polypeptide sequence specifically set forth herein.  
       [0152] A polypeptide “variant,” as the term is used herein, is a polypeptide that typically differs from a polypeptide specifically disclosed herein in one or more substitutions, deletions, additions and/or insertions. Such variants may be naturally occurring or may be synthetically generated, for example, by modifying one or more of the above polypeptide sequences of the invention and evaluating their immunogenic activity as described herein and/or using any of a number of techniques well known in the art.  
       [0153] For example, certain illustrative variants of the polypeptides of the invention include those in which one or more portions, such as an N-terminal leader sequence or transmembrane domain, have been removed. Other illustrative variants include variants in which a small portion (e.g., 1-30 amino acids, preferably 5-15 amino acids) has been removed from the N- and/or C-terminal of the mature protein.  
       [0154] In many instances, a variant will contain conservative substitutions. A “conservative substitution” is one in which an amino acid is substituted for another amino acid that has similar properties, such that one skilled in the art of peptide chemistry would expect the secondary structure and hydropathic nature of the polypeptide to be substantially unchanged. As described above, modifications may be made in the structure of the polynucleotides and polypeptides of the present invention and still obtain a functional molecule that encodes a variant or derivative polypeptide with desirable characteristics, e.g., with immunogenic characteristics. When it is desired to alter the amino acid sequence of a polypeptide to create an equivalent, or even an improved, immunogenic variant or portion of a polypeptide of the invention, one skilled in the art will typically change one or more of the codons of the encoding DNA sequence according to Table 1.  
       [0155] For example, certain amino acids may be substituted for other amino acids in a protein structure without appreciable loss of interactive binding capacity with structures such as, for example, antigen-binding regions of antibodies or binding sites on substrate molecules. Since it is the interactive capacity and nature of a protein that defines that protein&#39;s biological functional activity, certain amino acid sequence substitutions can be made in a protein sequence, and, of course, its underlying DNA coding sequence, and nevertheless obtain a protein with like properties. It is thus contemplated that various changes may be made in the peptide sequences of the disclosed compositions, or corresponding DNA sequences which encode said peptides without appreciable loss of their biological utility or activity.  
                   TABLE 1                       Amino Acids   Codons                                                                    Alanine   Ala   A   GCA   GCC   GCG   GCU               Cysteine   Cys   C   UGC   UGU       Aspartic acid   Asp   D   GAC   GAU       Glutamic acid   Glu   E   GAA   GAG       Phenylalanine   Phe   F   UUC   UUU       Glycine   Gly   G   GGA   GGC   GGG   GGU       Histidine   His   H   CAC   CAU       Isoleucine   Ile   I   AUA   AUC   AUU       Lysine   Lys   K   AAA   AAG       Leucine   Leu   L   UUA   UUG   CUA   CUC   CUG   CUU       Methionine   Met   M   AUG       Asparagine   Asn   N   AAC   AAU       Proline   Pro   P   CCA   CCC   CCG   CCU       Glutamine   Gln   Q   CAA   CAG       Arginine   Arg   R   AGA   AGG   CGA   CGC   CGG   CGU       Serine   Ser   S   AGC   AGU   UCA   UCC   UCG   UCU       Threonine   Thr   T   ACA   ACC   ACG   ACU       Valine   Val   V   GUA   GUC   GUG   GUU       Tryptophan   Trp   W   UGG       Tyrosine   Tyr   Y   UAC   UAU                  
 
       [0156] In making such changes, the hydropathic index of amino acids may be considered. The importance of the hydropathic amino acid index in conferring interactive biologic function on a protein is generally understood in the art (Kyte and Doolittle, 1982, incorporated herein by reference). It is accepted that the relative hydropathic character of the amino acid contributes to the secondary structure of the resultant protein, which in turn defines the interaction of the protein with other molecules, for example, enzymes, substrates, receptors, DNA, antibodies, antigens, and the like. Each amino acid has been assigned a hydropathic index on the basis of its hydrophobicity and charge characteristics (Kyte and Doolittle, 1982). These values are: isoleucine (+4.5); valine (+4.2); leucine (+3.8); phenylalanine (+2.8); cysteine/cystine (+2.5); methionine (+1.9); alanine (+1.8); glycine (−0.4); threonine (−0.7); serine (−0.8); tryptophan (−0.9); tyrosine (−1.3); proline (−1.6); histidine (−3.2); glutamate (-3.5); glutamine (−3.5); aspartate (−3.5); asparagine (−3.5); lysine (−3.9); and arginine (−4.5).  
       [0157] It is known in the art that certain amino acids may be substituted by other amino acids having a similar hydropathic index or score and still result in a protein with similar biological activity, i.e. still obtain a biological functionally equivalent protein. In making such changes, the substitution of amino acids whose hydropathic indices are within ±2 is preferred, those within ±1 are particularly preferred, and those within ±0.5 are even more particularly preferred. It is also understood in the art that the substitution of like amino acids can be made effectively on the basis of hydrophilicity. U.S. Pat. No. 4,554,101 (specifically incorporated herein by reference in its entirety), states that the greatest local average hydrophilicity of a protein, as governed by the hydrophilicity of its adjacent amino acids, correlates with a biological property of the protein.  
       [0158] As detailed in U.S. Pat. No. 4,554,101, the following hydrophilicity values have been assigned to amino acid residues: arginine (+3.0); lysine (+3.0); aspartate (+3.0±1); glutamate (+3.0±1); serine (+0.3); asparagine (+0.2); glutamine (+0.2); glycine (0); threonine (−0.4); proline (−0.5±1); alanine (−0.5); histidine (−0.5); cysteine (−1.0); methionine (−1.3); valine (−1.5); leucine (−1.8); isoleucine (−1.8); tyrosine (−2.3); phenylalanine (−2.5); tryptophan (−3.4). It is understood that an amino acid can be substituted for another having a similar hydrophilicity value and still obtain a biologically equivalent, and in particular, an immunologically equivalent protein. In such changes, the substitution of amino acids whose hydrophilicity values are within ±2 is preferred, those within ±1 are particularly preferred, and those within ±0.5 are even more particularly preferred.  
       [0159] As outlined above, amino acid substitutions are generally therefore based on the relative similarity of the amino acid side-chain substituents, for example, their hydrophobicity, hydrophilicity, charge, size, and the like. Exemplary substitutions that take various of the foregoing characteristics into consideration are well known to those of skill in the art and include: arginine and lysine; glutamate and aspartate; serine and threonine; glutamine and asparagine; and valine, leucine and isoleucine.  
       [0160] In addition, any polynucleotide may be further modified to increase stability in vivo. Possible modifications include, but are not limited to, the addition of flanking sequences at the 5′ and/or 3′ ends; the use of phosphorothioate or 2′ O-methyl rather than phosphodiesterase linkages in the backbone; and/or the inclusion of nontraditional bases such as inosine, queosine and wybutosine, as well as acetyl- methyl-, thio- and other modified forms of adenine, cytidine, guanine, thymine and uridine.  
       [0161] Amino acid substitutions may further be made on the basis of similarity in polarity, charge, solubility, hydrophobicity, hydrophilicity and/or the amphipathic nature of the residues. For example, negatively charged amino acids include aspartic acid and glutamic acid; positively charged amino acids include lysine and arginine; and amino acids with uncharged polar head groups having similar hydrophilicity values include leucine, isoleucine and valine; glycine and alanine; asparagine and glutamine; and serine, threonine, phenylalanine and tyrosine. Other groups of amino acids that may represent conservative changes include: (1) ala, pro, gly, glu, asp, gln, asn, ser, thr; (2) cys, ser, tyr, thr; (3) val, ile, leu, met, ala, phe; (4) lys, arg, his; and (5) phe, tyr, trp, his. A variant may also, or alternatively, contain nonconservative changes. In a preferred embodiment, variant polypeptides differ from a native sequence by substitution, deletion or addition of five amino acids or fewer. Variants may also (or alternatively) be modified by, for example, the deletion or addition of amino acids that have minimal influence on the immunogenicity, secondary structure and hydropathic nature of the polypeptide.  
       [0162] As noted above, polypeptides may comprise a signal (or leader) sequence at the N-terminal end of the protein, which co-translationally or post-translationally directs transfer of the protein. The polypeptide may also be conjugated to a linker or other sequence for ease of synthesis, purification or identification of the polypeptide (e.g., poly-His), or to enhance binding of the polypeptide to a solid support. For example, a polypeptide may be conjugated to an immunoglobulin Fc region.  
       [0163] When comparing polypeptide sequences, two sequences are said to be “identical” if the sequence of amino acids in the two sequences is the same when aligned for maximum correspondence, as described below. Comparisons between two sequences are typically performed by comparing the sequences over a comparison window to identify and compare local regions of sequence similarity. A “comparison window” as used herein, refers to a segment of at least about 20 contiguous positions, usually 30 to about 75, 40 to about 50, in which a sequence may be compared to a reference sequence of the same number of contiguous positions after the two sequences are optimally aligned.  
       [0164] Optimal alignment of sequences for comparison may be conducted using the Megalign program in the Lasergene suite of bioinformatics software (DNASTAR, Inc., Madison, Wis.), using default parameters. This program embodies several alignment schemes described in the following references: Dayhoff, M. O. (1978) A model of evolutionary change in proteins—Matrices for detecting distant relationships. In Dayhoff, M. O. (ed.) Atlas of Protein Sequence and Structure, National Biomedical Research Foundation, Washington D.C. Vol. 5, Suppl. 3, pp. 345-358; Hein J. (1990) Unified Approach to Alignment and Phylogenes pp. 626-645  Methods in Enzymology  vol. 183, Academic Press, Inc., San Diego, Calif.; Higgins, D. G. and Sharp, P. M. (1989)  CABIOS  5:151-153; Myers, E. W. and Muller W. (1988)  CABIOS  4:11-17; Robinson, E. D. (1971)  Comb. Theor  11:105; Saitou, N. Nei, M. (1987)  Mol. Biol. Evol.  4:406-425; Sneath, P. H. A. and Sokal, R. R. (1973)  Numerical Taxonomy—the Principles and Practice of Numerical Taxonomy,  Freeman Press, San Francisco, Calif.; Wilbur, W. J. and Lipman, D. J. (1983)  Proc. Natl. Acad., Sci. USA  80:726-730.  
       [0165] Alternatively, optimal alignment of sequences for comparison may be conducted by the local identity algorithm of Smith and Waterman (1981)  Add. APL. Math  2:482, by the identity alignment algorithm of Needleman and Wunsch (1970)  J. Mol. Biol.  48:443, by the search for similarity methods of Pearson and Lipman (1988)  Proc. Natl. Acad. Sci. USA  85: 2444, by computerized implementations of these algorithms (GAP, BESTFIT, BLAST, FASTA, and TFASTA in the Wisconsin Genetics Software Package, Genetics Computer Group (GCG), 575 Science Dr., Madison, Wis.), or by inspection.  
       [0166] One preferred example of algorithms that are suitable for determining percent sequence identity and sequence similarity are the BLAST and BLAST 2.0 algorithms, which are described in Altschul et al. (1977)  Nucl. Acids Res.  25:3389-3402 and Altschul et al. (1990)  J. Mol. Biol.  215:403-410, respectively. BLAST and BLAST 2.0 can be used, for example with the parameters described herein, to determine percent sequence identity for the polynucleotides and polypeptides of the invention. Software for performing BLAST analyses is publicly available through the National Center for Biotechnology Information. For amino acid sequences, a scoring matrix can be used to calculate the cumulative score. Extension of the word hits in each direction are halted when: the cumulative alignment score falls off by the quantity X from its maximum achieved value; the cumulative score goes to zero or below, due to the accumulation of one or more negative-scoring residue alignments; or the end of either sequence is reached. The BLAST algorithm parameters W, T and X determine the sensitivity and speed of the alignment.  
       [0167] In one preferred approach, the “percentage of sequence identity” is determined by comparing two optimally aligned sequences over a window of comparison of at least 20 positions, wherein the portion of the polypeptide sequence in the comparison window may comprise additions or deletions (ie., gaps) of 20 percent or less, usually 5 to 15 percent, or 10 to 12 percent, as compared to the reference sequences (which does not comprise additions or deletions) for optimal alignment of the two sequences. The percentage is calculated by determining the number of positions at which the identical amino acid residue occurs in both sequences to yield the number of matched positions, dividing the number of matched positions by the total number of positions in the reference sequence (i.e., the window size) and multiplying the results by 100 to yield the percentage of sequence identity.  
       [0168] Within other illustrative embodiments, a polypeptide may be a xenogeneic polypeptide that comprises an polypeptide having substantial sequence identity, as described above, to the human polypeptide (also termed autologous antigen) which served as a reference polypeptide, but which xenogeneic polypeptide is derived from a different, non-human species. One skilled in the art will recognize that “self” antigens are often poor stimulators of CD8+ and CD4+ T-lymphocyte responses, and therefore efficient immunotherapeutic strategies directed against tumor polypeptides require the development of methods to overcome immune tolerance to particular self tumor polypeptides. For example, humans immunized with prostase protein from a xenogeneic (non human) origin are capable of mounting an immune response against the counterpart human protein, e.g. the human prostase tumor protein present on human tumor cells. Accordingly, the present invention provides methods for purifying the xenogeneic form of the tumor proteins set forth herein, such as the polypeptides set forth in SEQ ID NO:36-41, 56, 57, 61, 62, 92 and 96, or those encoded by polynucleotide sequences set forth in SEQ ID NO: 1-35, 42-55, 58-60, 63-91 and 93-95.  
       [0169] Therefore, one aspect of the present invention provides xenogeneic variants of the polypeptide compositions described herein. Such xenogeneic variants generally encompassed by the present invention will typically exhibit at least about 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% or more identity along their lengths, to a polypeptide sequences set forth herein.  
       [0170] More particularly, the invention is directed to mouse, rat, monkey, porcine and other non-human polypeptides which can be used as xenogeneic forms of human polypeptides set forth herein, to induce immune responses directed against tumor polypeptides of the invention.  
       [0171] Within other illustrative embodiments, a polypeptide may be a fusion polypeptide that comprises multiple polypeptides as described herein, or that comprises at least one polypeptide as described herein and an unrelated sequence, such as a known tumor protein. A fusion partner may, for example, assist in providing T helper epitopes (an immunological fusion partner), preferably T helper epitopes recognized by humans, or may assist in expressing the protein (an expression enhancer) at higher yields than the native recombinant protein. Certain preferred fusion partners are both immunological and expression enhancing fusion partners. Other fusion partners may be selected so as to increase the solubility of the polypeptide or to enable the polypeptide to be targeted to desired intracellular compartments. Still further fusion partners include affinity tags, which facilitate purification of the polypeptide.  
       [0172] Fusion polypeptides may generally be prepared using standard techniques, including chemical conjugation. Preferably, a fusion polypeptide is expressed as a recombinant polypeptide, allowing the production of increased levels, relative to a non-fused polypeptide, in an expression system. Briefly, DNA sequences encoding the polypeptide components may be assembled separately, and ligated into an appropriate expression vector. The 3′ end of the DNA sequence encoding one polypeptide component is ligated, with or without a peptide linker, to the 5′ end of a DNA sequence encoding the second polypeptide component so that the reading frames of the sequences are in phase. This permits translation into a single fusion polypeptide that retains the biological activity of both component polypeptides.  
       [0173] A peptide linker sequence may be employed to separate the first and second polypeptide components by a distance sufficient to ensure that each polypeptide folds into its secondary and tertiary structures. Such a peptide linker sequence is incorporated into the fusion polypeptide using standard techniques well known in the art. Suitable peptide linker sequences may be chosen based on the following factors: (1) their ability to adopt a flexible extended conformation; (2) their inability to adopt a secondary structure that could interact with functional epitopes on the first and second polypeptides; and (3) the lack of hydrophobic or charged residues that might react with the polypeptide functional epitopes. Preferred peptide linker sequences contain Gly, Asn and Ser residues. Other near neutral amino acids, such as Thr and Ala may also be used in the linker sequence. Amino acid sequences which may be usefully employed as linkers include those disclosed in Maratea et al.,  Gene  40:39-46, 1985; Murphy et al.,  Proc. Natl. Acad. Sci. USA  83:8258-8262, 1986; U.S. Pat. No. 4,935,233 and U.S. Pat. No. 4,751,180. The linker sequence may generally be from 1 to about 50 amino acids in length. Linker sequences are not required when the first and second polypeptides have non-essential N-terminal amino acid regions that can be used to separate the functional domains and prevent steric interference.  
       [0174] The ligated DNA sequences are operably linked to suitable transcriptional or translational regulatory elements. The regulatory elements responsible for expression of DNA are located only 5′ to the DNA sequence encoding the first polypeptides. Similarly, stop codons required to end translation and transcription termination signals are only present 3′ to the DNA sequence encoding the second polypeptide.  
       [0175] The fusion polypeptide can comprise a polypeptide as described herein together with an unrelated immunogenic protein, such as an immunogenic protein capable of eliciting a recall response. Examples of such proteins include tetanus, tuberculosis and hepatitis proteins (see, for example, Stoute et al.  New Engl. J Med.,  336:86-91, 1997).  
       [0176] In one preferred embodiment, the immunological fusion partner is derived from a Mycobacterium sp., such as a Mycobacterium tuberculosis-derived Ral2 fragment. Ral2 compositions and methods for their use in enhancing the expression and/or immunogenicity of heterologous polynucleotide/polypeptide sequences is described in U.S. patent application Ser. No. 60/158,585, the disclosure of which is incorporated herein by reference in its entirety. Briefly, Ral2 refers to a polynucleotide region that is a subsequence of a Mycobacterium tuberculosis MTB32A nucleic acid. MTB32A is a serine protease of 32 KD molecular weight encoded by a gene in virulent and avirulent strains of  M. tuberculosis.  The nucleotide sequence and amino acid sequence of MTB32A have been described (for example, U.S. patent application Ser. No. 60/158,585; see also, Skeiky et al.,  Infection and Immun.  (1999) 67:3998-4007, incorporated herein by reference). C-terminal fragments of the MTB32A coding sequence express at high levels and remain as a soluble polypeptides throughout the purification process. Moreover, Ral2 may enhance the immunogenicity of heterologous immunogenic polypeptides with which it is fused. One preferred Ral2 fusion polypeptide comprises a 14 KD C-terminal fragment corresponding to amino acid residues 192 to 323 of MTB32A. Other preferred Ral2 polynucleotides generally comprise at least about 15 consecutive nucleotides, at least about 30 nucleotides, at least about 60 nucleotides, at least about 100 nucleotides, at least about 200 nucleotides, or at least about 300 nucleotides that encode a portion of a Ral2 polypeptide. Ral2 polynucleotides may comprise a native sequence (i.e., an endogenous sequence that encodes a Ral2 polypeptide or a portion thereof) or may comprise a variant of such a sequence. Ral2 polynucleotide variants may contain one or more substitutions, additions, deletions and/or insertions such that the biological activity of the encoded fusion polypeptide is not substantially diminished, relative to a fusion polypeptide comprising a native Ral2 polypeptide. Variants preferably exhibit at least about 70% identity, more preferably at least about 80% identity and most preferably at least about 90% identity to a polynucleotide sequence that encodes a native Ral2 polypeptide or a portion thereof.  
       [0177] Within other preferred embodiments, an immunological fusion partner is derived from protein D, a surface protein of the gram-negative bacterium Haemophilus influenza B (WO 91/18926). Preferably, a protein D derivative comprises approximately the first third of the protein (e.g., the first N-terminal 100-110 amino acids), and a protein D derivative may be lipidated. Within certain preferred embodiments, the first 109 residues of a Lipoprotein D fusion partner is included on the N-terminus to provide the polypeptide with additional exogenous T-cell epitopes and to increase the expression level in  E. coli  (thus functioning as an expression enhancer). The lipid tail ensures optimal presentation of the antigen to antigen presenting cells. Other fusion partners include the non-structural protein from influenzae virus, NS1 (hemaglutinin). Typically, the N-terminal 81 amino acids are used, although different fragments that include T-helper epitopes may be used.  
       [0178] In another embodiment, the immunological fusion partner is the protein known as LYTA, or a portion thereof (preferably a C-terminal portion). LYTA is derived from  Streptococcus pneumoniae,  which synthesizes an N-acetyl-L-alanine amidase known as amidase LYTA (encoded by the LytA gene;  Gene  43:265-292, 1986). LYTA is an autolysin that specifically degrades certain bonds in the peptidoglycan backbone. The C-terminal domain of the LYTA protein is responsible for the affinity to the choline or to some choline analogues such as DEAE. This property has been exploited for the development of  E. coli  C-LYTA expressing plasmids useful for expression of fusion proteins. Purification of hybrid proteins containing the C-LYTA fragment at the amino terminus has been described (see  Biotechnology  10:795-798, 1992). Within a preferred embodiment, a repeat portion of LYTA may be incorporated into a fusion polypeptide. A repeat portion is found in the C-terminal region starting at residue 178. A particularly preferred repeat portion incorporates residues 188-305.  
       [0179] Yet another illustrative embodiment involves fusion polypeptides, and the polynucleotides encoding them, wherein the fusion partner comprises a targeting signal capable of directing a polypeptide to the endosomal/lysosomal compartment, as described in U.S. Pat. No. 5,633,234. An immunogenic polypeptide of the invention, when fused with this targeting signal, will associate more efficiently with MHC class II molecules and thereby provide enhanced in vivo stimulation of CD4 +  T-cells specific for the polypeptide.  
       [0180] Polypeptides of the invention are prepared using any of a variety of well known synthetic and/or recombinant techniques, the latter of which are further described below. Polypeptides, portions and other variants generally less than about 150 amino acids can be generated by synthetic means, using techniques well known to those of ordinary skill in the art. In one illustrative example, such polypeptides are synthesized using any of the commercially available solid-phase techniques, such as the Merrifield solid-phase synthesis method, where amino acids are sequentially added to a growing amino acid chain. See Merrifield,  J. Am. Chem. Soc.  85:2149-2146, 1963. Equipment for automated synthesis of polypeptides is commercially available from suppliers such as Perkin Elmer/Applied BioSystems Division (Foster City, Calif.), and may be operated according to the manufacturer&#39;s instructions.  
       [0181] In general, polypeptide compositions (including fusion polypeptides) of the invention are isolated. An “isolated” polypeptide is one that is removed from its original environment. For example, a naturally-occurring protein or polypeptide is isolated if it is separated from some or all of the coexisting materials in the natural system. Preferably, such polypeptides are also purified, e.g., are at least about 90% pure, more preferably at least about 95% pure and most preferably at least about 99% pure.  
       [0182] Polynucleotide Compositions  
       [0183] The present invention, in other aspects, provides polynucleotide compositions. The terms “DNA” and “polynucleotide” are used essentially interchangeably herein to refer to a DNA molecule that has been isolated free of total genomic DNA of a particular species. “Isolated,” as used herein, means that a polynucleotide is substantially away from other coding sequences, and that the DNA molecule does not contain large portions of unrelated coding DNA, such as large chromosomal fragments or other functional genes or polypeptide coding regions. Of course, this refers to the DNA molecule as originally isolated, and does not exclude genes or coding regions later added to the segment by the hand of man.  
       [0184] As will be understood by those skilled in the art, the polynucleotide compositions of this invention can include genomic sequences, extra-genomic and plasmid-encoded sequences and smaller engineered gene segments that express, or may be adapted to express, proteins, polypeptides, peptides and the like. Such segments may be naturally isolated, or modified synthetically by the hand of man.  
       [0185] As will be also recognized by the skilled artisan, polynucleotides of the invention may be single-stranded (coding or antisense) or double-stranded, and may be DNA (genomic, cDNA or synthetic) or RNA molecules. RNA molecules may include HnRNA molecules, which contain introns and correspond to a DNA molecule in a one-to-one manner, and mRNA molecules, which do not contain introns. Additional coding or non-coding sequences may, but need not, be present within a polynucleotide of the present invention, and a polynucleotide may, but need not, be linked to other molecules and/or support materials.  
       [0186] Polynucleotides may comprise a native sequence (i.e., an endogenous sequence that encodes a polypeptide/protein of the invention or a portion thereof) or may comprise a sequence that encodes a variant or derivative, preferably and immunogenic variant or derivative, of such a sequence.  
       [0187] Therefore, according to another aspect of the present invention, polynucleotide compositions are provided that comprise some or all of a polynucleotide sequence set forth in any one of SEQ ID NO:1-35, 42-55, 58-60, 63-91 and 93-95, complements of a polynucleotide sequence set forth in any one of SEQ ID NO: 1-35, 42-55, 58-60, 63-91 and 93-95, and degenerate variants of a polynucleotide sequence set forth in any one of SEQ ID NO:1-35, 42-55, 58-60, 63-91 and 93-95. In certain preferred embodiments, the polynucleotide sequences set forth herein encode immunogenic polypeptides, as described above.  
       [0188] In other related embodiments, the present invention provides polynucleotide variants having substantial identity to the sequences disclosed herein in SEQ ID NO:1-35, 42-55, 58-60, 63-91 and 93-95, for example those comprising at least 70% sequence identity, preferably at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% or higher, sequence identity compared to a polynucleotide sequence of this invention using the methods described herein, (e.g., BLAST analysis using standard parameters, as described below). One skilled in this art will recognize that these values can be appropriately adjusted to determine corresponding identity of proteins encoded by two nucleotide sequences by taking into account codon degeneracy, amino acid similarity, reading frame positioning and the like.  
       [0189] Typically, polynucleotide variants will contain one or more substitutions, additions, deletions and/or insertions, preferably such that the immunogenicity of the polypeptide encoded by the variant polynucleotide is not substantially diminished relative to a polypeptide encoded by a polynucleotide sequence specifically set forth herein). The term “variants” should also be understood to encompasses homologous genes of xenogenic origin.  
       [0190] In additional embodiments, the present invention provides polynucleotide fragments comprising or consisting of various lengths of contiguous stretches of sequence identical to or complementary to one or more of the sequences disclosed herein. For example, polynucleotides are provided by this invention that comprise or consist of at least about 10, 15, 20, 30, 40, 50, 75, 100, 150, 200, 300, 400, 500 or 1000 or more contiguous nucleotides of one or more of the sequences disclosed herein as well as all intermediate lengths there between. It will be readily understood that “intermediate lengths”, in this context, means any length between the quoted values, such as 16, 17, 18, 19, etc.; 21, 22, 23, etc.; 30, 31, 32, etc.; 50, 51, 52, 53, etc.; 100, 101, 102, 103, etc.; 150, 151, 152, 153, etc.; including all integers through 200-500; 500-1,000, and the like. A polynucleotide sequence as described here may be extended at one or both ends by additional nucleotides not found in the native sequence. This additional sequence may consist of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 nucleotides at either end of the disclosed sequence or at both ends of the disclosed sequence.  
       [0191] In another embodiment of the invention, polynucleotide compositions are provided that are capable of hybridizing under moderate to high stringency conditions to a polynucleotide sequence provided herein, or a fragment thereof, or a complementary sequence thereof. Hybridization techniques are well known in the art of molecular biology. For purposes of illustration, suitable moderately stringent conditions for testing the hybridization of a polynucleotide of this invention with other polynucleotides include prewashing in a solution of 5×SSC, 0.5% SDS, 1.0 mM EDTA (pH 8.0); hybridizing at 50° C.-60° C., 5×SSC, overnight; followed by washing twice at 65° C. for 20 minutes with each of 2×, 0.5×and 0.2×SSC containing 0.1% SDS. One skilled in the art will understand that the stringency of hybridization can be readily manipulated, such as by altering the salt content of the hybridization solution and/or the temperature at which the hybridization is performed. For example, in another embodiment, suitable highly stringent hybridization conditions include those described above, with the exception that the temperature of hybridization is increased, e.g., to 60-65° C. or 65-70° C.  
       [0192] In certain preferred embodiments, the polynucleotides described above, e.g., polynucleotide variants, fragments and hybridizing sequences, encode polypeptides that are immunologically cross-reactive with a polypeptide sequence specifically set forth herein. In other preferred embodiments, such polynucleotides encode polypeptides that have a level of immunogenic activity of at least about 50%, preferably at least about 70%, and more preferably at least about 90% of that for a polypeptide sequence specifically set forth herein.  
       [0193] The polynucleotides of the present invention, or fragments thereof, regardless of the length of the coding sequence itself, may be combined with other DNA sequences, such as promoters, polyadenylation signals, additional restriction enzyme sites, multiple cloning sites, other coding segments, and the like, such that their overall length may vary considerably. It is therefore contemplated that a nucleic acid fragment of almost any length may be employed, with the total length preferably being limited by the ease of preparation and use in the intended recombinant DNA protocol. For example, illustrative polynucleotide segments with total lengths of about 10,000, about 5000, about 3000, about 2,000, about 1,000, about 500, about 200, about 100, about 50 base pairs in length, and the like, (including all intermediate lengths) are contemplated to be useful in many implementations of this invention.  
       [0194] When comparing polynucleotide sequences, two sequences are said to be “identical” if the sequence of nucleotides in the two sequences is the same when aligned for maximum correspondence, as described below. Comparisons between two sequences are typically performed by comparing the sequences over a comparison window to identify and compare local regions of sequence similarity. A “comparison window” as used herein, refers to a segment of at least about 20 contiguous positions, usually 30 to about 75, 40 to about 50, in which a sequence may be compared to a reference sequence of the same number of contiguous positions after the two sequences are optimally aligned.  
       [0195] Optimal alignment of sequences for comparison may be conducted using the Megalign program in the Lasergene suite of bioinformatics software (DNASTAR, Inc., Madison, Wis.), using default parameters. This program embodies several alignment schemes described in the following references: Dayhoff, M. O. (1978) A model of evolutionary change in proteins—Matrices for detecting distant relationships. In Dayhoff, M. O. (ed.) Atlas of Protein Sequence and Structure, National Biomedical Research Foundation, Washington D.C. Vol. 5, Suppl. 3, pp. 345-358; Hein J. (1990) Unified Approach to Alignment and Phylogenes pp. 626-645  Methods in Enzymology  vol. 183, Academic Press, Inc., San Diego, Calif.; Higgins, D. G. and Sharp, P. M. (1989)  CABIOS  5:151-153; Myers, E. W. and Muller W. (1988)  CABIOS  4:11-17; Robinson, E. D. (1971)  Comb. Theor  11:105; Santou, N. Nes, M. (1987)  Mol. Biol. Evol.  4:406-425; Sneath, P. H. A. and Sokal, R. R. (1973)  Numerical Taxonomy—the Principles and Practice of Numerical Taxonomy,  Freeman Press, San Francisco, Calif.; Wilbur, W. J. and Lipman, D. J. (1983)  Proc. Natl. Acad., Sci. USA  80:726-730.  
       [0196] Alternatively, optimal alignment of sequences for comparison may be conducted by the local identity algorithm of Smith and Waterman (1981)  Add. APL. Math  2:482, by the identity alignment algorithm of Needleman and Wunsch (1970)  J. Mol. Biol.  48:443, by the search for similarity methods of Pearson and Lipman (1988)  Proc. Natl. Acad. Sci. USA  85: 2444, by computerized implementations of these algorithms (GAP, BESTFIT, BLAST, FASTA, and TFASTA in the Wisconsin Genetics Software Package, Genetics Computer Group (GCG), 575 Science Dr., Madison, Wis.), or by inspection.  
       [0197] One preferred example of algorithms that are suitable for determining percent sequence identity and sequence similarity are the BLAST and BLAST 2.0 algorithms, which are described in Altschul et al. (1977)  Nucl. Acids Res.  25:3389-3402 and Altschul et al. (1990)  J. Mol. Biol.  215:403-410, respectively. BLAST and BLAST 2.0 can be used, for example with the parameters described herein, to determine percent sequence identity for the polynucleotides of the invention. Software for performing BLAST analyses is publicly available through the National Center for Biotechnology Information. In one illustrative example, cumulative scores can be calculated using, for nucleotide sequences, the parameters M (reward score for a pair of matching residues; always &gt;0) and N (penalty score for mismatching residues; always &lt;0). Extension of the word hits in each direction are halted when: the cumulative alignment score falls off by the quantity X from its maximum achieved value; the cumulative score goes to zero or below, due to the accumulation of one or more negative-scoring residue alignments; or the end of either sequence is reached. The BLAST algorithm parameters W, T and X determine the sensitivity and speed of the alignment. The BLASTN program (for nucleotide sequences) uses as defaults a wordlength (W) of 11, and expectation (E) of 10, and the BLOSUM62 scoring matrix (see Henikoff and Henikoff (1989)  Proc. Natl. Acad. Sci. USA  89:10915) alignments, (B) of 50, expectation (E) of 10, M=5, N=−4 and a comparison of both strands.  
       [0198] Preferably, the “percentage of sequence identity” is determined by comparing two optimally aligned sequences over a window of comparison of at least 20 positions, wherein the portion of the polynucleotide sequence in the comparison window may comprise additions or deletions (i.e., gaps) of 20 percent or less, usually 5 to 15 percent, or 10 to 12 percent, as compared to the reference sequences (which does not comprise additions or deletions) for optimal alignment of the two sequences. The percentage is calculated by determining the number of positions at which the identical nucleic acid bases occurs in both sequences to yield the number of matched positions, dividing the number of matched positions by the total number of positions in the reference sequence (i.e., the window size) and multiplying the results by 100 to yield the percentage of sequence identity.  
       [0199] It will be appreciated by those of ordinary skill in the art that, as a result of the degeneracy of the genetic code, there are many nucleotide sequences that encode a polypeptide as described herein. Some of these polynucleotides bear minimal homology to the nucleotide sequence of any native gene. Nonetheless, polynucleotides that vary due to differences in codon usage are specifically contemplated by the present invention. Further, alleles of the genes comprising the polynucleotide sequences provided herein are within the scope of the present invention. Alleles are endogenous genes that are altered as a result of one or more mutations, such as deletions, additions and/or substitutions of nucleotides. The resulting mRNA and protein may, but need not, have an altered structure or function. Alleles may be identified using standard techniques (such as hybridization, amplification and/or database sequence comparison).  
       [0200] Therefore, in another embodiment of the invention, a mutagenesis approach, such as site-specific mutagenesis, is employed for the preparation of immunogenic variants and/or derivatives of the polypeptides described herein. By this approach, specific modifications in a polypeptide sequence can be made through mutagenesis of the underlying polynucleotides that encode them. These techniques provides a straightforward approach to prepare and test sequence variants, for example, incorporating one or more of the foregoing considerations, by introducing one or more nucleotide sequence changes into the polynucleotide.  
       [0201] Site-specific mutagenesis allows the production of mutants through the use of specific oligonucleotide sequences which encode the DNA sequence of the desired mutation, as well as a sufficient number of adjacent nucleotides, to provide a primer sequence of sufficient size and sequence complexity to form a stable duplex on both sides of the deletion junction being traversed. Mutations may be employed in a selected polynucleotide sequence to improve, alter, decrease, modify, or otherwise change the properties of the polynucleotide itself, and/or alter the properties, activity, composition, stability, or primary sequence of the encoded polypeptide.  
       [0202] In certain embodiments of the present invention, the inventors contemplate the mutagenesis of the disclosed polynucleotide sequences to alter one or more properties of the encoded polypeptide, such as the immunogenicity of a polypeptide vaccine. The techniques of site-specific mutagenesis are well-known in the art, and are widely used to create variants of both polypeptides and polynucleotides. For example, site-specific mutagenesis is often used to alter a specific portion of a DNA molecule. In such embodiments, a primer comprising typically about 14 to about 25 nucleotides or so in length is employed, with about 5 to about 10 residues on both sides of the junction of the sequence being altered.  
       [0203] As will be appreciated by those of skill in the art, site-specific mutagenesis techniques have often employed a phage vector that exists in both a single stranded and double stranded form. Typical vectors useful in site-directed mutagenesis include vectors such as the M13 phage. These phage are readily commercially-available and their use is generally well-known to those skilled in the art. Double-stranded plasmids are also routinely employed in site directed mutagenesis that eliminates the step of transferring the gene of interest from a plasmid to a phage.  
       [0204] In general, site-directed mutagenesis in accordance herewith is performed by first obtaining a single-stranded vector or melting apart of two strands of a double-stranded vector that includes within its sequence a DNA sequence that encodes the desired peptide. An oligonucleotide primer bearing the desired mutated sequence is prepared, generally synthetically. This primer is then annealed with the single-stranded vector, and subjected to DNA polymerizing enzymes such as  E. coli  polymerase I Klenow fragment, in order to complete the synthesis of the mutation-bearing strand. Thus, a heteroduplex is formed wherein one strand encodes the original non-mutated sequence and the second strand bears the desired mutation. This heteroduplex vector is then used to transform appropriate cells, such as  E. coli  cells, and clones are selected which include recombinant vectors bearing the mutated sequence arrangement.  
       [0205] The preparation of sequence variants of the selected peptide-encoding DNA segments using site-directed mutagenesis provides a means of producing potentially useful species and is not meant to be limiting as there are other ways in which sequence variants of peptides and the DNA sequences encoding them may be obtained. For example, recombinant vectors encoding the desired peptide sequence may be treated with mutagenic agents, such as hydroxylamine, to obtain sequence variants. Specific details regarding these methods and protocols are found in the teachings of Maloy et al., 1994; Segal, 1976; Prokop and Bajpai, 1991; Kuby, 1994; and Maniatis et al., 1982, each incorporated herein by reference, for that purpose.  
       [0206] As used herein, the term “oligonucleotide directed mutagenesis procedure” refers to template-dependent processes and vector-mediated propagation which result in an increase in the concentration of a specific nucleic acid molecule relative to its initial concentration, or in an increase in the concentration of a detectable signal, such as amplification. As used herein, the term “oligonucleotide directed mutagenesis procedure” is intended to refer to a process that involves the template-dependent extension of a primer molecule. The term template dependent process refers to nucleic acid synthesis of a RNA or a DNA molecule wherein the sequence of the newly synthesized strand of nucleic acid is dictated by the well-known rules of complementary base pairing (see, for example, Watson, 1987). Typically, vector mediated methodologies involve the introduction of the nucleic acid fragment into a DNA or RNA vector, the clonal amplification of the vector, and the recovery of the amplified nucleic acid fragment. Examples of such methodologies are provided by U.S. Pat. No. 4,237,224, specifically incorporated herein by reference in its entirety.  
       [0207] In another approach for the production of polypeptide variants of the present invention, recursive sequence recombination, as described in U.S. Pat. No. 5,837,458, may be employed. In this approach, iterative cycles of recombination and screening or selection are performed to “evolve” individual polynucleotide variants of the invention having, for example, enhanced immunogenic activity.  
       [0208] In other embodiments of the present invention, the polynucleotide sequences provided herein can be advantageously used as probes or primers for nucleic acid hybridization. As such, it is contemplated that nucleic acid segments that comprise or consist of a sequence region of at least about a 15 nucleotide long contiguous sequence that has the same sequence as, or is complementary to, a 15 nucleotide long contiguous sequence disclosed herein will find particular utility. Longer contiguous identical or complementary sequences, e.g., those of about 20, 30, 40, 50, 100, 200, 500, 1000 (including all intermediate lengths) and even up to full length sequences will also be of use in certain embodiments.  
       [0209] The ability of such nucleic acid probes to specifically hybridize to a sequence of interest will enable them to be of use in detecting the presence of complementary sequences in a given sample. However, other uses are also envisioned, such as the use of the sequence information for the preparation of mutant species primers, or primers for use in preparing other genetic constructions.  
       [0210] Polynucleotide molecules having sequence regions consisting of contiguous nucleotide stretches of 10-14, 15-20, 30, 50, or even of 100-200 nucleotides or so (including intermediate lengths as well), identical or complementary to a polynucleotide sequence disclosed herein, are particularly contemplated as hybridization probes for use in, e.g., Southern and Northern blotting. This would allow a gene product, or fragment thereof, to be analyzed, both in diverse cell types and also in various bacterial cells. The total size of fragment, as well as the size of the complementary stretch(es), will ultimately depend on the intended use or application of the particular nucleic acid segment. Smaller fragments will generally find use in hybridization embodiments, wherein the length of the contiguous complementary region may be varied, such as between about 15 and about 100 nucleotides, but larger contiguous complementarity stretches may be used, according to the length complementary sequences one wishes to detect.  
       [0211] The use of a hybridization probe of about 15-25 nucleotides in length allows the formation of a duplex molecule that is both stable and selective. Molecules having contiguous complementary sequences over stretches greater than 15 bases in length are generally preferred, though, in order to increase stability and selectivity of the hybrid, and thereby improve the quality and degree of specific hybrid molecules obtained. One will generally prefer to design nucleic acid molecules having gene-complementary stretches of 15 to 25 contiguous nucleotides, or even longer where desired.  
       [0212] Hybridization probes may be selected from any portion of any of the sequences disclosed herein. All that is required is to review the sequences set forth herein, or to any continuous portion of the sequences, from about 15-25 nucleotides in length up to and including the full length sequence, that one wishes to utilize as a probe or primer. The choice of probe and primer sequences may be governed by various factors. For example, one may wish to employ primers from towards the termini of the total sequence.  
       [0213] Small polynucleotide segments or fragments may be readily prepared by, for example, directly synthesizing the fragment by chemical means, as is commonly practiced using an automated oligonucleotide synthesizer. Also, fragments may be obtained by application of nucleic acid reproduction technology, such as the PCR™ technology of U.S. Pat. No. 4,683,202 (incorporated herein by reference), by introducing selected sequences into recombinant vectors for recombinant production, and by other recombinant DNA techniques generally known to those of skill in the art of molecular biology.  
       [0214] The nucleotide sequences of the invention may be used for their ability to selectively form duplex molecules with complementary stretches of the entire gene or gene fragments of interest. Depending on the application envisioned, one will typically desire to employ varying conditions of hybridization to achieve varying degrees of selectivity of probe towards target sequence. For applications requiring high selectivity, one will typically desire to employ relatively stringent conditions to form the hybrids, e.g., one will select relatively low salt and/or high temperature conditions, such as provided by a salt concentration of from about 0.02 M to about 0.15 M salt at temperatures of from about 50° C. to about 70° C. Such selective conditions tolerate little, if any, mismatch between the probe and the template or target strand, and would be particularly suitable for isolating related sequences.  
       [0215] Of course, for some applications, for example, where one desires to prepare mutants employing a mutant primer strand hybridized to an underlying template, less stringent (reduced stringency) hybridization conditions will typically be needed in order to allow formation of the heteroduplex. In these circumstances, one may desire to employ salt conditions such as those of from about 0.15 M to about 0.9 M salt, at temperatures ranging from about 20° C. to about 55° C. Cross-hybridizing species can thereby be readily identified as positively hybridizing signals with respect to control hybridizations. In any case, it is generally appreciated that conditions can be rendered more stringent by the addition of increasing amounts of formamide, which serves to destabilize the hybrid duplex in the same manner as increased temperature. Thus, hybridization conditions can be readily manipulated, and thus will generally be a method of choice depending on the desired results.  
       [0216] According to another embodiment of the present invention, polynucleotide compositions comprising antisense oligonucleotides are provided. Antisense oligonucleotides have been demonstrated to be effective and targeted inhibitors of protein synthesis, and, consequently, provide a therapeutic approach by which a disease can be treated by inhibiting the synthesis of proteins that contribute to the disease. The efficacy of antisense oligonucleotides for inhibiting protein synthesis is well established. For example, the synthesis of polygalactauronase and the muscarine type 2 acetylcholine receptor are inhibited by antisense oligonucleotides directed to their respective mRNA sequences (U.S. Pat. No. 5,739,119 and U.S. Pat. No. 5,759,829). Further, examples of antisense inhibition have been demonstrated with the nuclear protein cyclin, the multiple drug resistance gene (MDG1), ICAM-1, E-selectin, STK-1, striatal GABA A  receptor and human EGF (Jaskulski et al., Science. 1988 Jun 10;240(4858):1544-6; Vasanthakumar and Ahmed, Cancer Commun. 1989;1(4):225-32; Peris et al., Brain Res Mol Brain Res. 1998 Jun 15;57(2):310-20; U.S. Pat. No. 5,801,154; U.S. Pat. No. 5,789,573; U.S. Pat. No. 5,718,709 and U.S. Pat. No. 5,610,288). Antisense constructs have also been described that inhibit and can be used to treat a variety of abnormal cellular proliferations, e.g. cancer (U.S. Pat. No. 5,747,470; U.S. Pat. No. 5,591,317 and U.S. Pat. No. 5,783,683).  
       [0217] Therefore, in certain embodiments, the present invention provides oligonucleotide sequences that comprise all, or a portion of, any sequence that is capable of specifically binding to polynucleotide sequence described herein, or a complement thereof. In one embodiment, the antisense oligonucleotides comprise DNA or derivatives thereof. In another embodiment, the oligonucleotides comprise RNA or derivatives thereof. In a third embodiment, the oligonucleotides are modified DNAs comprising a phosphorothioated modified backbone. In a fourth embodiment, the oligonucleotide sequences comprise peptide nucleic acids or derivatives thereof. In each case, preferred compositions comprise a sequence region that is complementary, and more preferably substantially-complementary, and even more preferably, completely complementary to one or more portions of polynucleotides disclosed herein. Selection of antisense compositions specific for a given gene sequence is based upon analysis of the chosen target sequence and determination of secondary structure, Tm, binding energy, and relative stability. Antisense compositions may be selected based upon their relative inability to form dimers, hairpins, or other secondary structures that would reduce or prohibit specific binding to the target mRNA in a host cell. Highly preferred target regions of the mRNA, are those which are at or near the AUG translation initiation codon, and those sequences which are substantially complementary to 5′ regions of the mRNA. These secondary structure analyses and target site selection considerations can be performed, for example, using v.4 of the OLIGO primer analysis software and/or the BLASTN 2.0.5 algorithm software (Altschul et al., Nucleic Acids Res. 1997, 25(17):3389-402).  
       [0218] The use of an antisense delivery method employing a short peptide vector, termed MPG (27 residues), is also contemplated. The MPG peptide contains a hydrophobic domain derived from the fusion sequence of HIV gp41 and a hydrophilic domain from the nuclear localization sequence of SV40 T-antigen (Morris et al., Nucleic Acids Res. 1997 Jul 15;25(14):2730-6). It has been demonstrated that several molecules of the MPG peptide coat the antisense oligonucleotides and can be delivered into cultured mammalian cells in less than 1 hour with relatively high efficiency (90%). Further, the interaction with MPG strongly increases both the stability of the oligonucleotide to nuclease and the ability to cross the plasma membrane.  
       [0219] According to another embodiment of the invention, the polynucleotide compositions described herein are used in the design and preparation of ribozyme molecules for inhibiting expression of the tumor polypeptides and proteins of the present invention in tumor cells. Ribozymes are RNA-protein complexes that cleave nucleic acids in a site-specific fashion. Ribozymes have specific catalytic domains that possess endonuclease activity (Kim and Cech, Proc Natl Acad Sci U S A. 1987 Dec;84(24):8788-92; Forster and Symons, Cell. 1987 Apr 24;49(2):211-20). For example, a large number of ribozymes accelerate phosphoester transfer reactions with a high degree of specificity, often cleaving only one of several phosphoesters in an oligonucleotide substrate (Cech et al., Cell. 1981 Dec;27(3 Pt 2):487-96; Michel and Westhof, J Mol Biol. 1990 Dec 5;216(3):585-610; Reinhold-Hurek and Shub, Nature. 1992 May 14;357(6374):173-6). This specificity has been attributed to the requirement that the substrate bind via specific base-pairing interactions to the internal guide sequence (“IGS”) of the ribozyme prior to chemical reaction.  
       [0220] Six basic varieties of naturally-occurring enzymatic RNAs are known presently. Each can catalyze the hydrolysis of RNA phosphodiester bonds in trans (and thus can cleave other RNA molecules) under physiological conditions. In general, enzymatic nucleic acids act by first binding to a target RNA. Such binding occurs through the target binding portion of a enzymatic nucleic acid which is held in close proximity to an enzymatic portion of the molecule that acts to cleave the target RNA. Thus, the enzymatic nucleic acid first recognizes and then binds a target RNA through complementary base-pairing, and once bound to the correct site, acts enzymatically to cut the target RNA. Strategic cleavage of such a target RNA will destroy its ability to direct synthesis of an encoded protein. After an enzymatic nucleic acid has bound and cleaved its RNA target, it is released from that RNA to search for another target and can repeatedly bind and cleave new targets.  
       [0221] The enzymatic nature of a ribozyme is advantageous over many technologies, such as antisense technology (where a nucleic acid molecule simply binds to a nucleic acid target to block its translation) since the concentration of ribozyme necessary to affect a therapeutic treatment is lower than that of an antisense oligonucleotide. This advantage reflects the ability of the ribozyme to act enzymatically. Thus, a single ribozyme molecule is able to cleave many molecules of target RNA. In addition, the ribozyme is a highly specific inhibitor, with the specificity of inhibition depending not only on the base pairing mechanism of binding to the target RNA, but also on the mechanism of target RNA cleavage. Single mismatches, or base-substitutions, near the site of cleavage can completely eliminate catalytic activity of a ribozyme. Similar mismatches in antisense molecules do not prevent their action (Woolf et al., Proc Natl Acad Sci U S A. 1992 Aug 15;89(16):7305-9). Thus, the specificity of action of a ribozyme is greater than that of an antisense oligonucleotide binding the same RNA site.  
       [0222] The enzymatic nucleic acid molecule may be formed in a hammerhead, hairpin, a hepatitis δ virus, group I intron or RNaseP RNA (in association with an RNA guide sequence) or Neurospora VS RNA motif. Examples of hammerhead motifs are described by Rossi et al. Nucleic Acids Res. 1992 Sep 11;20(17):4559-65. Examples of hairpin motifs are described by Hampel et al. (Eur. Pat. Appl. Publ. No. EP 0360257), Hampel and Tritz, Biochemistry 1989 Jun 13;28(12):4929-33; Hampel et al., Nucleic Acids Res. 1990 Jan 25;18(2):299-304 and U.S. Pat. No. 5,631,359. An example of the hepatitis δ virus motif is described by Perrotta and Been, Biochemistry. 1992 Dec 1;31(47):11843-52; an example of the RNaseP motif is described by Guerrier-Takada et al., Cell. 1983 Dec;35(3 Pt 2):849-57; Neurospora VS RNA ribozyme motif is described by Collins (Saville and Collins, Cell. 1990 May 18;61(4):685-96; Saville and Collins, Proc Natl Acad Sci U S A. 1991 Oct 1;88(19):8826-30; Collins and Olive, Biochemistry. 1993 Mar 23;32(11):2795-9); and an example of the Group I intron is described in (U.S. Pat. No. 4,987,071). All that is important in an enzymatic nucleic acid molecule of this invention is that it has a specific substrate binding site which is complementary to one or more of the target gene RNA regions, and that it have nucleotide sequences within or surrounding that substrate binding site which impart an RNA cleaving activity to the molecule. Thus the ribozyme constructs need not be limited to specific motifs mentioned herein.  
       [0223] Ribozymes may be designed as described in Int. Pat. Appl. Publ. No. WO 93/23569 and Int. Pat. Appl. Publ. No. WO 94/02595, each specifically incorporated herein by reference) and synthesized to be tested in vitro and in vivo, as described. Such ribozymes can also be optimized for delivery. While specific examples are provided, those in the art will recognize that equivalent RNA targets in other species can be utilized when necessary.  
       [0224] Ribozyme activity can be optimized by altering the length of the ribozyme binding arms, or chemically synthesizing ribozymes with modifications that prevent their degradation by serum ribonucleases (see e.g., Int. Pat. Appl. Publ. No. WO 92/07065; Int. Pat. Appl. Publ. No. WO 93/15187; Int. Pat. Appl. Publ. No. WO 91/03162; Eur. Pat. Appl. Publ. No. 92110298.4; U.S. Pat. No. 5,334,711; and Int. Pat. Appl. Publ. No. WO 94/13688, which describe various chemical modifications that can be made to the sugar moieties of enzymatic RNA molecules), modifications which enhance their efficacy in cells, and removal of stem II bases to shorten RNA synthesis times and reduce chemical requirements.  
       [0225] Sullivan et al. (Int. Pat. Appl. Publ. No. WO 94/02595) describes the general methods for delivery of enzymatic RNA molecules. Ribozymes may be administered to cells by a variety of methods known to those familiar to the art, including, but not restricted to, encapsulation in liposomes, by iontophoresis, or by incorporation into other vehicles, such as hydrogels, cyclodextrins, biodegradable nanocapsules, and bioadhesive microspheres. For some indications, ribozymes may be directly delivered ex vivo to cells or tissues with or without the aforementioned vehicles. Alternatively, the RNA/vehicle combination may be locally delivered by direct inhalation, by direct injection or by use of a catheter, infusion pump or stent. Other routes of delivery include, but are not limited to, intravascular, intramuscular, subcutaneous or joint injection, aerosol inhalation, oral (tablet or pill form), topical, systemic, ocular, intraperitoneal and/or intrathecal delivery. More detailed descriptions of ribozyme delivery and administration are provided in Int. Pat. Appl. Publ. No. WO 94/02595 and Int. Pat. Appl. Publ. No. WO 93/23569, each specifically incorporated herein by reference.  
       [0226] Another means of accumulating high concentrations of a ribozyme(s) within cells is to incorporate the ribozyme-encoding sequences into a DNA expression vector. Transcription of the ribozyme sequences are driven from a promoter for eukaryotic RNA polymerase I (pol I), RNA polymerase II (pol II), or RNA polymerase III (pol III). Transcripts from pol II or pol III promoters will be expressed at high levels in all cells; the levels of a given pol II promoter in a given cell type will depend on the nature of the gene regulatory sequences (enhancers, silencers, etc.) present nearby. Prokaryotic RNA polymerase promoters may also be used, providing that the prokaryotic RNA polymerase enzyme is expressed in the appropriate cells Ribozymes expressed from such promoters have been shown to function in mammalian cells. Such transcription units can be incorporated into a variety of vectors for introduction into mammalian cells, including but not restricted to, plasmid DNA vectors, viral DNA vectors (such as adenovirus or adeno-associated vectors), or viral RNA vectors (such as retroviral, semliki forest virus, sindbis virus vectors).  
       [0227] In another embodiment of the invention, peptide nucleic acids (PNAs) compositions are provided. PNA is a DNA mimic in which the nucleobases are attached to a pseudopeptide backbone (Good and Nielsen, Antisense Nucleic Acid Drug Dev. 1997 7(4) 431-37). PNA is able to be utilized in a number methods that traditionally have used RNA or DNA. Often PNA sequences perform better in techniques than the corresponding RNA or DNA sequences and have utilities that are not inherent to RNA or DNA. A review of PNA including methods of making, characteristics of, and methods of using, is provided by Corey ( Trends Biotechnol  1997 Jun;15(6):224-9). As such, in certain embodiments, one may prepare PNA sequences that are complementary to one or more portions of the ACE mRNA sequence, and such PNA compositions may be used to regulate, alter, decrease, or reduce the translation of ACE-specific mRNA, and thereby alter the level of ACE activity in a host cell to which such PNA compositions have been administered.  
       [0228] PNAs have 2-aminoethyl-glycine linkages replacing the normal phosphodiester backbone of DNA (Nielsen et al.,  Science  1991 Dec 6;254(5037):1497-500; Hanvey et al., Science. 1992 Nov 27;258(5087):1481-5; Hyrup and Nielsen, Bioorg Med Chem. 1996 Jan;4(1):5-23). This chemistry has three important consequences: firstly, in contrast to DNA or phosphorothioate oligonucleotides, PNAs are neutral molecules; secondly, PNAs are achiral, which avoids the need to develop a stereoselective synthesis; and thirdly, PNA synthesis uses standard Boc or Fmoc protocols for solid-phase peptide synthesis, although other methods, including a modified Merrifield method, have been used.  
       [0229] PNA monomers or ready-made oligomers are commercially available from PerSeptive Biosystems (Framingham, Mass.). PNA syntheses by either Boc or Fmoc protocols are straightforward using manual or automated protocols (Norton et al., Bioorg Med Chem. 1995 Apr;3(4):437-45). The manual protocol lends itself to the production of chemically modified PNAs or the simultaneous synthesis of families of closely related PNAs.  
       [0230] As with peptide synthesis, the success of a particular PNA synthesis will depend on the properties of the chosen sequence. For example, while in theory PNAs can incorporate any combination of nucleotide bases, the presence of adjacent purines can lead to deletions of one or more residues in the product. In expectation of this difficulty, it is suggested that, in producing PNAs with adjacent purines, one should repeat the coupling of residues likely to be added inefficiently. This should be followed by the purification of PNAs by reverse-phase high-pressure liquid chromatography, providing yields and purity of product similar to those observed during the synthesis of peptides.  
       [0231] Modifications of PNAs for a given application may be accomplished by coupling amino acids during solid-phase synthesis or by attaching compounds that contain a carboxylic acid group to the exposed N-terminal amine. Alternatively, PNAs can be modified after synthesis by coupling to an introduced lysine or cysteine. The ease with which PNAs can be modified facilitates optimization for better solubility or for specific functional requirements. Once synthesized, the identity of PNAs and their derivatives can be confirmed by mass spectrometry. Several studies have made and utilized modifications of PNAs (for example, Norton et al., Bioorg Med Chem. 1995 Apr;3(4):437-45; Petersen et al., J Pept Sci. 1995 May-Jun;1(3):175-83; Orum et al., Biotechniques. 1995 Sep;19(3):472-80; Footer et al., Biochemistry. 1996 Aug 20;35(33):10673-9; Griffith et al., Nucleic Acids Res. 1995 Aug 11;23(15):3003-8; Pardridge et al., Proc Natl Acad Sci U S A. 1995 Jun 6;92(12):5592-6; Boffa et al., Proc Natl Acad Sci U S A. 1995 Mar 14;92(6):1901-5; Gambacorti-Passerini et al., Blood. 1996 Aug 15;88(4):1411-7; Armitage et al., Proc Natl Acad Sci U S A. 1997 Nov 11;94(23):12320-5; Seeger et al., Biotechniques. 1997 Sep;23(3):512-7). U.S. Pat. No. 5,700,922 discusses PNA-DNA-PNA chimeric molecules and their uses in diagnostics, modulating protein in organisms, and treatment of conditions susceptible to therapeutics.  
       [0232] Methods of characterizing the antisense binding properties of PNAs are discussed in Rose (Anal Chem. 1993 Dec 15;65(24):3545-9) and Jensen et al. (Biochemistry. 1997 Apr 22;36(16):5072-7). Rose uses capillary gel electrophoresis to determine binding of PNAs to their complementary oligonucleotide, measuring the relative binding kinetics and stoichiometry. Similar types of measurements were made by Jensen et al. using BIAcore™ technology.  
       [0233] Other applications of PNAs that have been described and will be apparent to the skilled artisan include use in DNA strand invasion, antisense inhibition, mutational analysis, enhancers of transcription, nucleic acid purification, isolation of transcriptionally active genes, blocking of transcription factor binding, genome cleavage, biosensors, in situ hybridization, and the like.  
       [0234] Polynucleotide Identification, Characterization and Expression  
       [0235] Polynucleotides compositions of the present invention may be identified, prepared and/or manipulated using any of a variety of well established techniques (see generally, Sambrook et al.,  Molecular Cloning: A Laboratory Manual,  Cold Spring Harbor Laboratories, Cold Spring Harbor, N.Y., 1989, and other like references). For example, a polynucleotide may be identified, as described in more detail below, by screening a microarray of cDNAs for tumor-associated expression (i.e., expression that is at least two fold greater in a tumor than in normal tissue, as determined using a representative assay provided herein). Such screens may be performed, for example, using the microarray technology of Affymetrix, Inc. (Santa Clara, Calif.) according to the manufacturer&#39;s instructions (and essentially as described by Schena et al.,  Proc. Natl. Acad. Sci. USA  93:10614-10619, 1996 and Heller et al.,  Proc. Natl. Acad. Sci. USA  94:2150-2155, 1997). Alternatively, polynucleotides may be amplified from cDNA prepared from cells expressing the proteins described herein, such as tumor cells.  
       [0236] Many template dependent processes are available to amplify a target sequences of interest present in a sample. One of the best known amplification methods is the polymerase chain reaction (PCR™) which is described in detail in U.S. Pat. Nos. 4,683,195, 4,683,202 and 4,800,159, each of which is incorporated herein by reference in its entirety. Briefly, in PCR™, two primer sequences are prepared which are complementary to regions on opposite complementary strands of the target sequence. An excess of deoxynucleoside triphosphates is added to a reaction mixture along with a DNA polymerase (e.g., Taq polymerase). If the target sequence is present in a sample, the primers will bind to the target and the polymerase will cause the primers to be extended along the target sequence by adding on nucleotides. By raising and lowering the temperature of the reaction mixture, the extended primers will dissociate from the target to form reaction products, excess primers will bind to the target and to the reaction product and the process is repeated. Preferably reverse transcription and PCR™ amplification procedure may be performed in order to quantify the amount of mRNA amplified. Polymerase chain reaction methodologies are well known in the art.  
       [0237] Any of a number of other template dependent processes, many of which are variations of the PCR™ amplification technique, are readily known and available in the art. Illustratively, some such methods include the ligase chain reaction (referred to as LCR), described, for example, in Eur. Pat. Appl. Publ. No. 320,308 and U.S. Pat. No. 4,883,750; Qbeta Replicase, described in PCT Intl. Pat. Appl. Publ. No. PCT/US87/00880; Strand Displacement Amplification (SDA) and Repair Chain Reaction (RCR). Still other amplification methods are described in Great Britain Pat. Appl. No. 2 202 328, and in PCT Intl. Pat. Appl. Publ. No. PCT/US89/01025. Other nucleic acid amplification procedures include transcription-based amplification systems (TAS) (PCT Intl. Pat. Appl. Publ. No. WO 88/10315), including nucleic acid sequence based amplification (NASBA) and 3SR. Eur. Pat. Appl. Publ. No. 329,822 describes a nucleic acid amplification process involving cyclically synthesizing single-stranded RNA (“ssRNA”), ssDNA, and double-stranded DNA (dsDNA). PCT Intl. Pat. Appl. Publ. No. WO 89/06700 describes a nucleic acid sequence amplification scheme based on the hybridization of a promoter/primer sequence to a target single-stranded DNA (“ssDNA”) followed by transcription of many RNA copies of the sequence. Other amplification methods such as “RACE” (Frohman, 1990), and “one-sided PCR” (Ohara, 1989) are also well-known to those of skill in the art.  
       [0238] An amplified portion of a polynucleotide of the present invention may be used to isolate a full length gene from a suitable library (e.g., a tumor cDNA library) using well known techniques. Within such techniques, a library (cDNA or genomic) is screened using one or more polynucleotide probes or primers suitable for amplification. Preferably, a library is size-selected to include larger molecules. Random primed libraries may also be preferred for identifying 5′ and upstream regions of genes. Genomic libraries are preferred for obtaining introns and extending 5′ sequences.  
       [0239] For hybridization techniques, a partial sequence may be labeled (e.g., by nick-translation or end-labeling with  32 P) using well known techniques. A bacterial or bacteriophage library is then generally screened by hybridizing filters containing denatured bacterial colonies (or lawns containing phage plaques) with the labeled probe (see Sambrook et al.,  Molecular Cloning: A Laboratory Manual,  Cold Spring Harbor Laboratories, Cold Spring Harbor, N.Y., 1989). Hybridizing colonies or plaques are selected and expanded, and the DNA is isolated for further analysis. cDNA clones may be analyzed to determine the amount of additional sequence by, for example, PCR using a primer from the partial sequence and a primer from the vector. Restriction maps and partial sequences may be generated to identify one or more overlapping clones. The complete sequence may then be determined using standard techniques, which may involve generating a series of deletion clones. The resulting overlapping sequences can then assembled into a single contiguous sequence. A full length cDNA molecule can be generated by ligating suitable fragments, using well known techniques.  
       [0240] Alternatively, amplification techniques, such as those described above, can be useful for obtaining a full length coding sequence from a partial cDNA sequence. One such amplification technique is inverse PCR (see Triglia et al.,  Nucl. Acids Res.  16:8186, 1988), which uses restriction enzymes to generate a fragment in the known region of the gene. The fragment is then circularized by intramolecular ligation and used as a template for PCR with divergent primers derived from the known region. Within an alternative approach, sequences adjacent to a partial sequence may be retrieved by amplification with a primer to a linker sequence and a primer specific to a known region. The amplified sequences are typically subjected to a second round of amplification with the same linker primer and a second primer specific to the known region. A variation on this procedure, which employs two primers that initiate extension in opposite directions from the known sequence, is described in WO 96/38591. Another such technique is known as “rapid amplification of cDNA ends” or RACE. This technique involves the use of an internal primer and an external primer, which hybridizes to a polyA region or vector sequence, to identify sequences that are 5′ and 3′ of a known sequence. Additional techniques include capture PCR (Lagerstrom et al.,  PCR Methods Applic.  1:111-19, 1991) and walking PCR (Parker et al.,  Nucl. Acids. Res.  19:3055-60, 1991). Other methods employing amplification may also be employed to obtain a full length cDNA sequence.  
       [0241] In certain instances, it is possible to obtain a full length cDNA sequence by analysis of sequences provided in an expressed sequence tag (EST) database, such as that available from GenBank. Searches for overlapping ESTs may generally be performed using well known programs (e.g., NCBI BLAST searches), and such ESTs may be used to generate a contiguous full length sequence. Full length DNA sequences may also be obtained by analysis of genomic fragments.  
       [0242] In other embodiments of the invention, polynucleotide sequences or fragments thereof which encode polypeptides of the invention, or fusion proteins or functional equivalents thereof, may be used in recombinant DNA molecules to direct expression of a polypeptide in appropriate host cells. Due to the inherent degeneracy of the genetic code, other DNA sequences that encode substantially the same or a functionally equivalent amino acid sequence may be produced and these sequences may be used to clone and express a given polypeptide.  
       [0243] As will be understood by those of skill in the art, it may be advantageous in some instances to produce polypeptide-encoding nucleotide sequences possessing non-naturally occurring codons. For example, codons preferred by a particular prokaryotic or eukaryotic host can be selected to increase the rate of protein expression or to produce a recombinant RNA transcript having desirable properties, such as a half-life which is longer than that of a transcript generated from the naturally occurring sequence.  
       [0244] Moreover, the polynucleotide sequences of the present invention can be engineered using methods generally known in the art in order to alter polypeptide encoding sequences for a variety of reasons, including but not limited to, alterations which modify the cloning, processing, and/or expression of the gene product. For example, DNA shuffling by random fragmentation and PCR reassembly of gene fragments and synthetic oligonucleotides may be used to engineer the nucleotide sequences. In addition, site-directed mutagenesis may be used to insert new restriction sites, alter glycosylation patterns, change codon preference, produce splice variants, or introduce mutations, and so forth.  
       [0245] In another embodiment of the invention, natural, modified, or recombinant nucleic acid sequences may be ligated to a heterologous sequence to encode a fusion protein. For example, to screen peptide libraries for inhibitors of polypeptide activity, it may be useful to encode a chimeric protein that can be recognized by a commercially available antibody. A fusion protein may also be engineered to contain a cleavage site located between the polypeptide-encoding sequence and the heterologous protein sequence, so that the polypeptide may be cleaved and purified away from the heterologous moiety.  
       [0246] Sequences encoding a desired polypeptide may be synthesized, in whole or in part, using chemical methods well known in the art (see Caruthers, M. H. et al. (1980)  Nucl. Acids Res. Symp. Ser.  215-223, Horn, T. et al. (1980)  Nucl. Acids Res. Symp. Ser.  225-232). Alternatively, the protein itself may be produced using chemical methods to synthesize the amino acid sequence of a polypeptide, or a portion thereof. For example, peptide synthesis can be performed using various solid-phase techniques (Roberge, J. Y. et al. (1995)  Science  269:202-204) and automated synthesis may be achieved, for example, using the ABI 431A Peptide Synthesizer (Perkin Elmer, Palo Alto, Calif.).  
       [0247] A newly synthesized peptide may be substantially purified by preparative high performance liquid chromatography (e.g., Creighton, T. (1983) Proteins, Structures and Molecular Principles, WH Freeman and Co., New York, N.Y.) or other comparable techniques available in the art. The composition of the synthetic peptides may be confirmed by amino acid analysis or sequencing (e.g., the Edman degradation procedure). Additionally, the amino acid sequence of a polypeptide, or any part thereof, may be altered during direct synthesis and/or combined using chemical methods with sequences from other proteins, or any part thereof, to produce a variant polypeptide.  
       [0248] In order to express a desired polypeptide, the nucleotide sequences encoding the polypeptide, or functional equivalents, may be inserted into appropriate expression vector, i.e., a vector which contains the necessary elements for the transcription and translation of the inserted coding sequence. Methods which are well known to those skilled in the art may be used to construct expression vectors containing sequences encoding a polypeptide of interest and appropriate transcriptional and translational control elements. These methods include in vitro recombinant DNA techniques, synthetic techniques, and in vivo genetic recombination. Such techniques are described, for example, in Sambrook, J. et al. (1989) Molecular Cloning, A Laboratory Manual, Cold Spring Harbor Press, Plainview, N.Y., and Ausubel, F. M. et al. (1989) Current Protocols in Molecular Biology, John Wiley &amp; Sons, New York. N.Y.  
       [0249] A variety of expression vector/host systems may be utilized to contain and express polynucleotide sequences. These include, but are not limited to, microorganisms such as bacteria transformed with recombinant bacteriophage, plasmid, or cosmid DNA expression vectors; yeast transformed with yeast expression vectors; insect cell systems infected with virus expression vectors (e.g., baculovirus); plant cell systems transformed with virus expression vectors (e.g., cauliflower mosaic virus, CaMV; tobacco mosaic virus, TMV) or with bacterial expression vectors (e.g., Ti or pBR322 plasmids); or animal cell systems.  
       [0250] The “control elements” or “regulatory sequences” present in an expression vector are those non-translated regions of the vector—enhancers, promoters, 5′ and 3′ untranslated regions—which interact with host cellular proteins to carry out transcription and translation. Such elements may vary in their strength and specificity. Depending on the vector system and host utilized, any number of suitable transcription and translation elements, including constitutive and inducible promoters, may be used. For example, when cloning in bacterial systems, inducible promoters such as the hybrid lacZ promoter of the pBLUESCRIPT phagemid (Stratagene, La Jolla, Calif.) or pSPORT1 plasmid (Gibco BRL, Gaithersburg, Md.) and the like may be used. In mammalian cell systems, promoters from mammalian genes or from mammalian viruses are generally preferred. If it is necessary to generate a cell line that contains multiple copies of the sequence encoding a polypeptide, vectors based on SV40 or EBV may be advantageously used with an appropriate selectable marker.  
       [0251] In bacterial systems, any of a number of expression vectors may be selected depending upon the use intended for the expressed polypeptide. For example, when large quantities are needed, for example for the induction of antibodies, vectors which direct high level expression of fusion proteins that are readily purified may be used. Such vectors include, but are not limited to, the multifunctional  E. coli  cloning and expression vectors such as pBLUESCRIPT (Stratagene), in which the sequence encoding the polypeptide of interest may be ligated into the vector in frame with sequences for the amino-terminal Met and the subsequent 7 residues of .beta.-galactosidase so that a hybrid protein is produced; pIN vectors (Van Heeke, G. and S. M. Schuster (1989)  J. Biol. Chem.  264:5503-5509); and the like. pGEX Vectors (Promega, Madison, Wis.) may also be used to express foreign polypeptides as fusion proteins with glutathione S-transferase (GST). In general, such fusion proteins are soluble and can easily be purified from lysed cells by adsorption to glutathione-agarose beads followed by elution in the presence of free glutathione. Proteins made in such systems may be designed to include heparin, thrombin, or factor XA protease cleavage sites so that the cloned polypeptide of interest can be released from the GST moiety at will.  
       [0252] In the yeast,  Saccharomyces cerevisiae,  a number of vectors containing constitutive or inducible promoters such as alpha factor, alcohol oxidase, and PGH may be used. For reviews, see Ausubel et al. (supra) and Grant et al. (1987)  Methods Enzymol.  153:516-544.  
       [0253] In cases where plant expression vectors are used, the expression of sequences encoding polypeptides may be driven by any of a number of promoters. For example, viral promoters such as the 35S and 19S promoters of CaMV may be used alone or in combination with the omega leader sequence from TMV (Takamatsu, N. (1987)  EMBO J.  6:307-311. Alternatively, plant promoters such as the small subunit of RUBISCO or heat shock promoters may be used (Coruzzi, G. et al. (1984)  EMBO J.  3:1671-1680; Broglie, R. et al. (1984)  Science  224:838-843; and Winter, J. et al. (1991)  Results Probl. Cell Differ.  17:85-105). These constructs can be introduced into plant cells by direct DNA transformation or pathogen-mediated transfection. Such techniques are described in a number of generally available reviews (see, for example, Hobbs, S. or Murry, L. E. in McGraw Hill Yearbook of Science and Technology (1992) McGraw Hill, New York, N.Y.; pp. 191-196).  
       [0254] An insect system may also be used to express a polypeptide of interest. For example, in one such system,  Autographa califormica  nuclear polyhedrosis virus (AcNPV) is used as a vector to express foreign genes in  Spodoptera frugiperda  cells or in  Trichoplusia larvae.  The sequences encoding the polypeptide may be cloned into a non-essential region of the virus, such as the polyhedrin gene, and placed under control of the polyhedrin promoter. Successful insertion of the polypeptide-encoding sequence will render the polyhedrin gene inactive and produce recombinant virus lacking coat protein. The recombinant viruses may then be used to infect, for example,  S. frugiperda  cells or  Trichoplusia larvae  in which the polypeptide of interest may be expressed (Engelhard, E. K. et al. (1994)  Proc. Natl. Acad. Sci.  91 :3224-3227).  
       [0255] In mammalian host cells, a number of viral-based expression systems are generally available. For example, in cases where an adenovirus is used as an expression vector, sequences encoding a polypeptide of interest may be ligated into an adenovirus transcription/translation complex consisting of the late promoter and tripartite leader sequence. Insertion in a non-essential E1 or E3 region of the viral genome may be used to obtain a viable virus which is capable of expressing the polypeptide in infected host cells (Logan, J. and Shenk, T. (1984)  Proc. Natl. Acad. Sci.  81:3655-3659). In addition, transcription enhancers, such as the Rous sarcoma virus (RSV) enhancer, may be used to increase expression in mammalian host cells.  
       [0256] Specific initiation signals may also be used to achieve more efficient translation of sequences encoding a polypeptide of interest. Such signals include the ATG initiation codon and adjacent sequences. In cases where sequences encoding the polypeptide, its initiation codon, and upstream sequences are inserted into the appropriate expression vector, no additional transcriptional or translational control signals may be needed. However, in cases where only coding sequence, or a portion thereof, is inserted, exogenous translational control signals including the ATG initiation codon should be provided. Furthermore, the initiation codon should be in the correct reading frame to ensure translation of the entire insert. Exogenous translational elements and initiation codons may be of various origins, both natural and synthetic. The efficiency of expression may be enhanced by the inclusion of enhancers which are appropriate for the particular cell system which is used, such as those described in the literature (Scharf, D. et al. (1994)  Results Probl. Cell Differ.  20:125-162).  
       [0257] In addition, a host cell strain may be chosen for its ability to modulate the expression of the inserted sequences or to process the expressed protein in the desired fashion. Such modifications of the polypeptide include, but are not limited to, acetylation, carboxylation. glycosylation, phosphorylation, lipidation, and acylation. Post-translational processing which cleaves a “prepro” form of the protein may also be used to facilitate correct insertion, folding and/or function. Different host cells such as CHO, COS, HeLa, MDCK, HEK293, and W138, which have specific cellular machinery and characteristic mechanisms for such post-translational activities, may be chosen to ensure the correct modification and processing of the foreign protein.  
       [0258] For long-term, high-yield production of recombinant proteins, stable expression is generally preferred. For example, cell lines which stably express a polynucleotide of interest may be transformed using expression vectors which may contain viral origins of replication and/or endogenous expression elements and a selectable marker gene on the same or on a separate vector. Following the introduction of the vector, cells may be allowed to grow for 1-2 days in an enriched media before they are switched to selective media. The purpose of the selectable marker is to confer resistance to selection, and its presence allows growth and recovery of cells which successfully express the introduced sequences. Resistant clones of stably transformed cells may be proliferated using tissue culture techniques appropriate to the cell type.  
       [0259] Any number of selection systems may be used to recover transformed cell lines. These include, but are not limited to, the herpes simplex virus thymidine kinase (Wigler, M. et al. (1977)  Cell  11:223-32) and adenine phosphoribosyltransferase (Lowy, I. et al. (1990)  Cell  22:817-23) genes which can be employed in tk.sup.- or aprt.sup.-cells, respectively. Also, antimetabolite, antibiotic or herbicide resistance can be used as the basis for selection; for example, dhfr which confers resistance to methotrexate (Wigler, M. et al. (1980)  Proc. Natl. Acad Sci.  77:3567-70); npt, which confers resistance to the aminoglycosides, neomycin and G-418 (Colbere-Garapin, F. et al (1981)  J. Mol. Biol.  150:1-14); and als or pat, which confer resistance to chlorsulfuron and phosphinotricin acetyltransferase, respectively (Murry, supra). Additional selectable genes have been described, for example, trpB, which allows cells to utilize indole in place of tryptophan, or hisD, which allows cells to utilize histinol in place of histidine (Hartman, S. C. and R. C. Mulligan (1988)  Proc. Natl. Acad. Sci.  85:8047-51). The use of visible markers has gained popularity with such markers as anthocyanins, beta-glucuronidase and its substrate GUS, and luciferase and its substrate luciferin, being widely used not only to identify transformants, but also to quantify the amount of transient or stable protein expression attributable to a specific vector system (Rhodes, C. A. et al. (1995)  Methods Mol. Biol.  55:121-131).  
       [0260] Although the presence/absence of marker gene expression suggests that the gene of interest is also present, its presence and expression may need to be confirmed. For example, if the sequence encoding a polypeptide is inserted within a marker gene sequence, recombinant cells containing sequences can be identified by the absence of marker gene function. Alternatively, a marker gene can be placed in tandem with a polypeptide-encoding sequence under the control of a single promoter. Expression of the marker gene in response to induction or selection usually indicates expression of the tandem gene as well.  
       [0261] Alternatively, host cells that contain and express a desired polynucleotide sequence may be identified by a variety of procedures known to those of skill in the art. These procedures include, but are not limited to, DNA-DNA or DNA-RNA hybridizations and protein bioassay or immunoassay techniques which include, for example, membrane, solution, or chip based technologies for the detection and/or quantification of nucleic acid or protein.  
       [0262] A variety of protocols for detecting and measuring the expression of polynucleotide-encoded products, using either polyclonal or monoclonal antibodies specific for the product are known in the art. Examples include enzyme-linked immunosorbent assay (ELISA), radioimmunoassay (RIA), and fluorescence activated cell sorting (FACS). A two-site, monoclonal-based immunoassay utilizing monoclonal antibodies reactive to two non-interfering epitopes on a given polypeptide may be preferred for some applications, but a competitive binding assay may also be employed. These and other assays are described, among other places, in Hampton, R. et al. (1990; Serological Methods, a Laboratory Manual, APS Press, St Paul. Minn.) and Maddox, D. E. et al. (1983;  J Exp. Med.  158:1211-1216).  
       [0263] A wide variety of labels and conjugation techniques are known by those skilled in the art and may be used in various nucleic acid and amino acid assays. Means for producing labeled hybridization or PCR probes for detecting sequences related to polynucleotides include oligolabeling, nick translation, end-labeling or PCR amplification using a labeled nucleotide. Alternatively, the sequences, or any portions thereof may be cloned into a vector for the production of an mRNA probe. Such vectors are known in the art, are commercially available, and may be used to synthesize RNA probes in vitro by addition of an appropriate RNA polymerase such as T7, T3, or SP6 and labeled nucleotides. These procedures may be conducted using a variety of commercially available kits. Suitable reporter molecules or labels, which may be used include radionuclides, enzymes, fluorescent, chemiluminescent, or chromogenic agents as well as substrates, cofactors, inhibitors, magnetic particles, and the like.  
       [0264] Host cells transformed with a polynucleotide sequence of interest may be cultured under conditions suitable for the expression and recovery of the protein from cell culture. The protein produced by a recombinant cell may be secreted or contained intracellularly depending on the sequence and/or the vector used. As will be understood by those of skill in the art, expression vectors containing polynucleotides of the invention may be designed to contain signal sequences which direct secretion of the encoded polypeptide through a prokaryotic or eukaryotic cell membrane. Other recombinant constructions may be used to join sequences encoding a polypeptide of interest to nucleotide sequence encoding a polypeptide domain which will facilitate purification of soluble proteins. Such purification facilitating domains include, but are not limited to, metal chelating peptides such as histidine-tryptophan modules that allow purification on immobilized metals, protein A domains that allow purification on immobilized immunoglobulin, and the domain utilized in the FLAGS extension/affinity purification system (Immunex Corp., Seattle, Wash.). The inclusion of cleavable linker sequences such as those specific for Factor XA or enterokinase (Invitrogen. San Diego, Calif.) between the purification domain and the encoded polypeptide may be used to facilitate purification. One such expression vector provides for expression of a fusion protein containing a polypeptide of interest and a nucleic acid encoding 6 histidine residues preceding a thioredoxin or an enterokinase cleavage site. The histidine residues facilitate purification on IMIAC (immobilized metal ion affinity chromatography) as described in Porath, J. et al. (1992,  Prot. Exp. Purif  3:263-281) while the enterokinase cleavage site provides a means for purifying the desired polypeptide from the fusion protein. A discussion of vectors which contain fusion proteins is provided in Kroll, D. J. et al. (1993;  DNA Cell Biol.  12:441-453).  
       [0265] In addition to recombinant production methods, polypeptides of the invention, and fragments thereof, may be produced by direct peptide synthesis using solid-phase techniques (Merrifield J. (1963)  J. Am. Chem. Soc.  85:2149-2154). Protein synthesis may be performed using manual techniques or by automation. Automated synthesis may be achieved, for example, using Applied Biosystems 431A Peptide Synthesizer (Perkin Elmer). Alternatively, various fragments may be chemically synthesized separately and combined using chemical methods to produce the full length molecule.  
       [0266] Antibody Compositions Fragments Thereof and Other Binding Agents  
       [0267] According to another aspect, the present invention further provides binding agents, such as antibodies and antigen-binding fragments thereof, that exhibit immunological binding to a tumor polypeptide disclosed herein, or to a portion, variant or derivative thereof. An antibody, or antigen-binding fragment thereof, is said to “specifically bind,” “immunogically bind,” and/or is “immunologically reactive” to a polypeptide of the invention if it reacts at a detectable level (within, for example, an ELISA assay) with the polypeptide, and does not react detectably with unrelated polypeptides under similar conditions.  
       [0268] Immunological binding, as used in this context, generally refers to the non-covalent interactions of the type which occur between an immunoglobulin molecule and an antigen for which the immunoglobulin is specific. The strength, or affinity of immunological binding interactions can be expressed in terms of the dissociation constant (K d ) of the interaction, wherein a smaller K d  represents a greater affinity. Immunological binding properties of selected polypeptides can be quantified using methods well known in the art. One such method entails measuring the rates of antigen-binding site/antigen complex formation and dissociation, wherein those rates depend on the concentrations of the complex partners, the affinity of the interaction, and on geometric parameters that equally influence the rate in both directions. Thus, both the “on rate constant” (K on ) and the “off rate constant” (K off ) can be determined by calculation of the concentrations and the actual rates of association and dissociation. The ratio of K off /K on  enables cancellation of all parameters not related to affinity, and is thus equal to the dissociation constant K d . See, generally, Davies et al. (1990) Annual Rev. Biochem. 59:439-473.  
       [0269] An “antigen-binding site,” or “binding portion” of an antibody refers to the part of the immunoglobulin molecule that participates in antigen binding. The antigen binding site is formed by amino acid residues of the N-terminal variable (“V”) regions of the heavy (“H”) and light (“L”) chains. Three highly divergent stretches within the V regions of the heavy and light chains are referred to as “hypervariable regions” which are interposed between more conserved flanking stretches known as “framework regions,” or “FRs”. Thus the term “FR” refers to amino acid sequences which are naturally found between and adjacent to hypervariable regions in immunoglobulins. In an antibody molecule, the three hypervariable regions of a light chain and the three hypervariable regions of a heavy chain are disposed relative to each other in three dimensional space to form an antigen-binding surface. The antigen-binding surface is complementary to the three-dimensional surface of a bound antigen, and the three hypervariable regions of each of the heavy and light chains are referred to as “complementarity-determining regions,” or “CDRs.” 
       [0270] Binding agents may be further capable of differentiating between patients with and without a cancer, such as lung cancer, using the representative assays provided herein. For example, antibodies or other binding agents that bind to a tumor protein will preferably generate a signal indicating the presence of a cancer in at least about 20% of patients with the disease, more preferably at least about 30% of patients. Alternatively, or in addition, the antibody will generate a negative signal indicating the absence of the disease in at least about 90% of individuals without the cancer. To determine whether a binding agent satisfies this requirement, biological samples (e.g., blood, sera, sputum, urine and/or tumor biopsies) from patients with and without a cancer (as determined using standard clinical tests) may be assayed as described herein for the presence of polypeptides that bind to the binding agent. Preferably, a statistically significant number of samples with and without the disease will be assayed. Each binding agent should satisfy the above criteria; however, those of ordinary skill in the art will recognize that binding agents may be used in combination to improve sensitivity.  
       [0271] Any agent that satisfies the above requirements may be a binding agent. For example, a binding agent may be a ribosome, with or without a peptide component, an RNA molecule or a polypeptide. In a preferred embodiment, a binding agent is an antibody or an antigen-binding fragment thereof. Antibodies may be prepared by any of a variety of techniques known to those of ordinary skill in the art. See, e.g., Harlow and Lane,  Antibodies: A Laboratory Manual,  Cold Spring Harbor Laboratory, 1988. In general, antibodies can be produced by cell culture techniques, including the generation of monoclonal antibodies as described herein, or via transfection of antibody genes into suitable bacterial or mammalian cell hosts, in order to allow for the production of recombinant antibodies. In one technique, an immunogen comprising the polypeptide is initially injected into any of a wide variety of mammals (e.g., mice, rats, rabbits, sheep or goats). In this step, the polypeptides of this invention may serve as the immunogen without modification. Alternatively, particularly for relatively short polypeptides, a superior immune response may be elicited if the polypeptide is joined to a carrier protein, such as bovine serum albumin or keyhole limpet hemocyanin. The immunogen is injected into the animal host, preferably according to a predetermined schedule incorporating one or more booster immunizations, and the animals are bled periodically. Polyclonal antibodies specific for the polypeptide may then be purified from such antisera by, for example, affinity chromatography using the polypeptide coupled to a suitable solid support.  
       [0272] Monoclonal antibodies specific for an antigenic polypeptide of interest may be prepared, for example, using the technique of Kohler and Milstein,  Eur. J Immunol.  6:511-519, 1976, and improvements thereto. Briefly, these methods involve the preparation of immortal cell lines capable of producing antibodies having the desired specificity (i.e., reactivity with the polypeptide of interest). Such cell lines may be produced, for example, from spleen cells obtained from an animal immunized as described above. The spleen cells are then immortalized by, for example, fusion with a myeloma cell fusion partner, preferably one that is syngeneic with the immunized animal. A variety of fusion techniques may be employed. For example, the spleen cells and myeloma cells may be combined with a nonionic detergent for a few minutes and then plated at low density on a selective medium that supports the growth of hybrid cells, but not myeloma cells. A preferred selection technique uses HAT (hypoxanthine, aminopterin, thymidine) selection. After a sufficient time, usually about 1 to 2 weeks, colonies of hybrids are observed. Single colonies are selected and their culture supernatants tested for binding activity against the polypeptide. Hybridomas having high reactivity and specificity are preferred.  
       [0273] Monoclonal antibodies may be isolated from the supernatants of growing hybridoma colonies. In addition, various techniques may be employed to enhance the yield, such as injection of the hybridoma cell line into the peritoneal cavity of a suitable vertebrate host, such as a mouse. Monoclonal antibodies may then be harvested from the ascites fluid or the blood. Contaminants may be removed from the antibodies by conventional techniques, such as chromatography, gel filtration, precipitation, and extraction. The polypeptides of this invention may be used in the purification process in, for example, an affinity chromatography step.  
       [0274] A number of therapeutically useful molecules are known in the art which comprise antigen-binding sites that are capable of exhibiting immunological binding properties of an antibody molecule. The proteolytic enzyme papain preferentially cleaves IgG molecules to yield several fragments, two of which (the “F(ab)” fragments) each comprise a covalent heterodimer that includes an intact antigen-binding site. The enzyme pepsin is able to cleave IgG molecules to provide several fragments, including the “F(ab′) 2  ” fragment which comprises both antigen-binding sites. An “Fv” fragment can be produced by preferential proteolytic cleavage of an IgM, and on rare occasions IgG or IgA immunoglobulin molecule. Fv fragments are, however, more commonly derived using recombinant techniques known in the art. The Fv fragment includes a non-covalent V H ::V L  heterodimer including an antigen-binding site which retains much of the antigen recognition and binding capabilities of the native antibody molecule. Inbar et al. (1972) Proc. Nat. Acad. Sci. USA 69:2659-2662; Hochman et al. (1976) Biochem 15:2706-2710; and Ehrlich et al. (1980) Biochem 19:4091-4096.  
       [0275] A single chain Fv (“sFv”) polypeptide is a covalently linked V H ::V L  heterodimer which is expressed from a gene fusion including V H - and V L -encoding genes linked by a peptide-encoding linker. Huston et al. (1988) Proc. Nat. Acad. Sci. USA 85(16):5879-5883. A number of methods have been described to discern chemical structures for converting the naturally aggregated—but chemically separated—light and heavy polypeptide chains from an antibody V region into an sFv molecule which will fold into a three dimensional structure substantially similar to the structure of an antigen-binding site. See, e.g., U.S. Pat. Nos. 5,091,513 and 5,132,405, to Huston et al.; and U.S. Pat. No. 4,946,778, to Ladner et al.  
       [0276] Each of the above-described molecules includes a heavy chain and a light chain CDR set, respectively interposed between a heavy chain and a light chain FR set which provide support to the CDRS and define the spatial relationship of the CDRs relative to each other. As used herein, the term “CDR set” refers to the three hypervariable regions of a heavy or light chain V region. Proceeding from the N-terminus of a heavy or light chain, these regions are denoted as “CDR1,” “CDR2,” and “CDR3” respectively. An antigen-binding site, therefore, includes six CDRs, comprising the CDR set from each of a heavy and a light chain V region. A polypeptide comprising a single CDR, (e.g., a CDR1, CDR2 or CDR3) is referred to herein as a “molecular recognition unit.” Crystallographic analysis of a number of antigen-antibody complexes has demonstrated that the amino acid residues of CDRs form extensive contact with bound antigen, wherein the most extensive antigen contact is with the heavy chain CDR3. Thus, the molecular recognition units are primarily responsible for the specificity of an antigen-binding site.  
       [0277] As used herein, the term “FR set” refers to the four flanking amino acid sequences which frame the CDRs of a CDR set of a heavy or light chain V region. Some FR residues may contact bound antigen; however, FRs are primarily responsible for folding the V region into the antigen-binding site, particularly the FR residues directly adjacent to the CDRS. Within FRs, certain amino residues and certain structural features are very highly conserved. In this regard, all V region sequences contain an internal disulfide loop of around 90 amino acid residues. When the V regions fold into a binding-site, the CDRs are displayed as projecting loop motifs which form an antigen-binding surface. It is generally recognized that there are conserved structural regions of FRs which influence the folded shape of the CDR loops into certain “canonical” structures—regardless of the precise CDR amino acid sequence. Further, certain FR residues are known to participate in non-covalent interdomain contacts which stabilize the interaction of the antibody heavy and light chains.  
       [0278] A number of “humanized” antibody molecules comprising an antigen-binding site derived from a non-human immunoglobulin have been described, including chimeric antibodies having rodent V regions and their associated CDRs fused to human constant domains (Winter et al. (1991) Nature 349:293-299; Lobuglio et al. (1989) Proc. Nat. Acad. Sci. USA 86:4220-4224; Shaw et al. (1987) J Immunol. 138:4534-4538; and Brown et al. (1987) Cancer Res. 47:3577-3583), rodent CDRs grafted into a human supporting FR prior to fusion with an appropriate human antibody constant domain (Riechmann et al. (1988) Nature 332:323-327; Verhoeyen et al. (1988) Science 239:1534-1536; and Jones et al. (1986) Nature 321:522-525), and rodent CDRs supported by recombinantly veneered rodent FRs (European Patent Publication No. 519,596, published Dec. 23, 1992). These “humanized” molecules are designed to minimize unwanted immunological response toward rodent antihuman antibody molecules which limits the duration and effectiveness of therapeutic applications of those moieties in human recipients.  
       [0279] As used herein, the terms “veneered FRs” and “recombinantly veneered FRs” refer to the selective replacement of FR residues from, e.g., a rodent heavy or light chain V region, with human FR residues in order to provide a xenogeneic molecule comprising an antigen-binding site which retains substantially all of the native FR polypeptide folding structure. Veneering techniques are based on the understanding that the ligand binding characteristics of an antigen-binding site are determined primarily by the structure and relative disposition of the heavy and light chain CDR sets within the antigen-binding surface. Davies et al. (1990) Ann. Rev. Biochem. 59:439-473. Thus, antigen binding specificity can be preserved in a humanized antibody only wherein the CDR structures, their interaction with each other, and their interaction with the rest of the V region domains are carefully maintained. By using veneering techniques, exterior (e.g., solvent-accessible) FR residues which are readily encountered by the immune system are selectively replaced with human residues to provide a hybrid molecule that comprises either a weakly immunogenic, or substantially non-immunogenic veneered surface.  
       [0280] The process of veneering makes use of the available sequence data for human antibody variable domains compiled by Kabat et al., in Sequences of Proteins of Immunological Interest, 4th ed., (U.S. Dept. of Health and Human Services, U.S. Government Printing Office, 1987), updates to the Kabat database, and other accessible U.S. and foreign databases (both nucleic acid and protein). Solvent accessibilities of V region amino acids can be deduced from the known three-dimensional structure for human and murine antibody fragments. There are two general steps in veneering a murine antigen-binding site. Initially, the FRs of the variable domains of an antibody molecule of interest are compared with corresponding FR sequences of human variable domains obtained from the above-identified sources. The most homologous human V regions are then compared residue by residue to corresponding murine amino acids. The residues in the murine FR which differ from the human counterpart are replaced by the residues present in the human moiety using recombinant techniques well known in the art. Residue switching is only carried out with moieties which are at least partially exposed (solvent accessible), and care is exercised in the replacement of amino acid residues which may have a significant effect on the tertiary structure of V region domains, such as proline, glycine and charged amino acids.  
       [0281] In this manner, the resultant “veneered” murine antigen-binding sites are thus designed to retain the murine CDR residues, the residues substantially adjacent to the CDRs, the residues identified as buried or mostly buried (solvent inaccessible), the residues believed to participate in non-covalent (e.g., electrostatic and hydrophobic) contacts between heavy and light chain domains, and the residues from conserved structural regions of the FRs which are believed to influence the “canonical” tertiary structures of the CDR loops. These design criteria are then used to prepare recombinant nucleotide sequences which combine the CDRs of both the heavy and light chain of a murine antigen-binding site into human-appearing FRs that can be used to transfect mammalian cells for the expression of recombinant human antibodies which exhibit the antigen specificity of the murine antibody molecule.  
       [0282] In another embodiment of the invention, monoclonal antibodies of the present invention may be coupled to one or more therapeutic agents. Suitable agents in this regard include radionuclides, differentiation inducers, drugs, toxins, and derivatives thereof. Preferred radionuclides include  90 Y,  123 I,  125 I,  131 I,  186 Re,  188 Re,  211 At, and  212 Bi. Preferred drugs include methotrexate, and pyrimidine and purine analogs. Preferred differentiation inducers include phorbol esters and butyric acid. Preferred toxins include ricin, abrin, diptheria toxin, cholera toxin, gelonin, Pseudomonas exotoxin, Shigella toxin, and pokeweed antiviral protein.  
       [0283] A therapeutic agent may be coupled (e.g., covalently bonded) to a suitable monoclonal antibody either directly or indirectly (e.g., via a linker group). A direct reaction between an agent and an antibody is possible when each possesses a substituent capable of reacting with the other. For example, a nucleophilic group, such as an amino or sulfhydryl group, on one may be capable of reacting with a carbonyl-containing group, such as an anhydride or an acid halide, or with an alkyl group containing a good leaving group (e.g., a halide) on the other.  
       [0284] Alternatively, it may be desirable to couple a therapeutic agent and an antibody via a linker group. A linker group can function as a spacer to distance an antibody from an agent in order to avoid interference with binding capabilities. A linker group can also serve to increase the chemical reactivity of a substituent on an agent or an antibody, and thus increase the coupling efficiency. An increase in chemical reactivity may also facilitate the use of agents, or functional groups on agents, which otherwise would not be possible.  
       [0285] It will be evident to those skilled in the art that a variety of bifunctional or polyfunctional reagents, both homo- and hetero-functional (such as those described in the catalog of the Pierce Chemical Co., Rockford, Ill.), may be employed as the linker group. Coupling may be effected, for example, through amino groups, carboxyl groups, sulfhydryl groups or oxidized carbohydrate residues. There are numerous references describing such methodology, e.g., U.S. Pat. No. 4,671,958, to Rodwell et al.  
       [0286] Where a therapeutic agent is more potent when free from the antibody portion of the immunoconjugates of the present invention, it may be desirable to use a linker group which is cleavable during or upon internalization into a cell. A number of different cleavable linker groups have been described. The mechanisms for the intracellular release of an agent from these linker groups include cleavage by reduction of a disulfide bond (e.g., U.S. Pat. No. 4,489,710, to Spitler), by irradiation of a photolabile bond (e.g., U.S. Pat. No. 4,625,014, to Senter et al.), by hydrolysis of derivatized amino acid side chains (e.g., U.S. Pat. No. 4,638,045, to Kohn et al.), by serum complement-mediated hydrolysis (e.g., U.S. Pat. No. 4,671,958, to Rodwell et al.), and acid-catalyzed hydrolysis (e.g., U.S. Pat. No. 4,569,789, to Blattler et al.).  
       [0287] It may be desirable to couple more than one agent to an antibody. In one embodiment, multiple molecules of an agent are coupled to one antibody molecule. In another embodiment, more than one type of agent may be coupled to one antibody. Regardless of the particular embodiment, immunoconjugates with more than one agent may be prepared in a variety of ways. For example, more than one agent may be coupled directly to an antibody molecule, or linkers that provide multiple sites for attachment can be used. Alternatively, a carrier can be used.  
       [0288] A carrier may bear the agents in a variety of ways, including covalent bonding either directly or via a linker group. Suitable carriers include proteins such as albumins (e.g., U.S. Pat. No. 4,507,234, to Kato et al.), peptides and polysaccharides such as aminodextran (e.g., U.S. Pat. No. 4,699,784, to Shih et al.). A carrier may also bear an agent by noncovalent bonding or by encapsulation, such as within a liposome vesicle (e.g., U.S. Pat. Nos. 4,429,008 and 4,873,088). Carriers specific for radionuclide agents include radiohalogenated small molecules and chelating compounds. For example, U.S. Pat. No. 4,735,792 discloses representative radiohalogenated small molecules and their synthesis. A radionuclide chelate may be formed from chelating compounds that include those containing nitrogen and sulfur atoms as the donor atoms for binding the metal, or metal oxide, radionuclide. For example, U.S. Pat. No. 4,673,562, to Davison et al. discloses representative chelating compounds and their synthesis.  
       [0289] T Cell Compositions  
       [0290] The present invention, in another aspect, provides T cells specific for a tumor polypeptide disclosed herein, or for a variant or derivative thereof. Such cells may generally be prepared in vitro or ex vivo, using standard procedures. For example, T cells may be isolated from bone marrow, peripheral blood, or a fraction of bone marrow or peripheral blood of a patient, using a commercially available cell separation system, such as the Isolex™ System, available from Nexell Therapeutics, Inc. (Irvine, Calif.; see also U.S. Pat. No. 5,240,856; U.S. Pat. No. 5,215,926; WO 89/06280; WO 91/16116 and WO 92/07243). Alternatively, T cells may be derived from related or unrelated humans, non-human mammals, cell lines or cultures.  
       [0291] T cells may be stimulated with a polypeptide, polynucleotide encoding a polypeptide and/or an antigen presenting cell (APC) that expresses such a polypeptide. Such stimulation is performed under conditions and for a time sufficient to permit the generation of T cells that are specific for the polypeptide of interest. Preferably, a tumor polypeptide or polynucleotide of the invention is present within a delivery vehicle, such as a microsphere, to facilitate the generation of specific T cells.  
       [0292] T cells are considered to be specific for a polypeptide of the present invention if the T cells specifically proliferate, secrete cytokines or kill target cells coated with the polypeptide or expressing a gene encoding the polypeptide. T cell specificity may be evaluated using any of a variety of standard techniques. For example, within a chromium release assay or proliferation assay, a stimulation index of more than two fold increase in lysis and/or proliferation, compared to negative controls, indicates T cell specificity. Such assays may be performed, for example, as described in Chen et al.,  Cancer Res.  54:1065-1070, 1994. Alternatively, detection of the proliferation of T cells may be accomplished by a variety of known techniques. For example, T cell proliferation can be detected by measuring an increased rate of DNA synthesis (e.g., by pulse-labeling cultures of T cells with tritiated thymidine and measuring the amount of tritiated thymidine incorporated into DNA). Contact with a tumor polypeptide (100 ng/ml -100 μg/ml, preferably 200 ng/ml -25 μg/ml) for 3-7 days will typically result in at least a two fold increase in proliferation of the T cells. Contact as described above for 2-3 hours should result in activation of the T cells, as measured using standard cytokine assays in which a two fold increase in the level of cytokine release (e.g., TNF or IFN-γ) is indicative of T cell activation (see Coligan et al., Current Protocols in Immunology, vol. 1, Wiley Interscience (Greene 1998)). T cells that have been activated in response to a tumor polypeptide, polynucleotide or polypeptide-expressing APC may be CD4 +  and/or CD8 + . Tumor polypeptide-specific T cells may be expanded using standard techniques. Within preferred embodiments, the T cells are derived from a patient, a related donor or an unrelated donor, and are administered to the patient following stimulation and expansion.  
       [0293] For therapeutic purposes, CD4 +  or CD8 +  T cells that proliferate in response to a tumor polypeptide, polynucleotide or APC can be expanded in number either in vitro or in vivo. Proliferation of such T cells in vitro may be accomplished in a variety of ways. For example, the T cells can be re-exposed to a tumor polypeptide, or a short peptide corresponding to an immunogenic portion of such a polypeptide, with or without the addition of T cell growth factors, such as interleukin-2, and/or stimulator cells that synthesize a tumor polypeptide. Alternatively, one or more T cells that proliferate in the presence of the tumor polypeptide can be expanded in number by cloning. Methods for cloning cells are well known in the art, and include limiting dilution.  
       [0294] T Cell Receptor Compositions  
       [0295] The T cell receptor (TCR) consists of 2 different, highly variable polypeptide chains, termed the T-cell receptor α and β chains, that are linked by a disulfide bond (Janeway, Travers, Walport.  Immunobiology.  Fourth Ed., 148-159. Elsevier Science Ltd/Garland Publishing. 1999). The α/β heterodimer complexes with the invariant CD3 chains at the cell membrane. This complex recognizes specific antigenic peptides bound to MHC molecules. The enormous diversity of TCR specificities is generated much like immunoglobulin diversity, through somatic gene rearrangement. The α chain genes contain over 50 variable (V), 2 diversity (D), over 10 joining (J) segments, and 2 constant region segments (C). The a chain genes contain over 70 V segments, and over 60 J segments but no D segments, as well as one C segment. During T cell development in the thymus, the D to J gene rearrangement of the β chain occurs, followed by the V gene segment rearrangement to the DJ. This functional VDJ β  exon is transcribed and spliced to join to a C β . For the α chain, a V α  gene segment rearranges to a J α  gene segment to create the functional exon that is then transcribed and spliced to the C α . Diversity is further increased during the recombination process by the random addition of P and N-nucleotides between the V, D, and J segments of the β chain and between the V and J segments in the a chain (Janeway, Travers, Walport.  Immunobiology.  Fourth Ed., 98 and 150. Elsevier Science Ltd/Garland Publishing. 1999).  
       [0296] The present invention, in another aspect, provides TCRs specific for a polypeptide disclosed herein, or for a variant or derivative thereof. In accordance with the present invention, polynucleotide and amino acid sequences are provided for the V-J or V-D-J junctional regions or parts thereof for the alpha and beta chains of the T-cell receptor which recognize tumor polypeptides described herein. In general, this aspect of the invention relates to T-cell receptors which recognize or bind tumor polypeptides presented in the context of MHC. In a preferred embodiment the tumor antigens recognized by the T-cell receptors comprise a polypeptide of the present invention. For example, cDNA encoding a TCR specific for a lung tumor peptide can be isolated from T cells specific for a tumor polypeptide using standard molecular biological and recombinant DNA techniques.  
       [0297] This invention further includes the T-cell receptors or analogs thereof having substantially the same function or activity as the T-cell receptors of this invention which recognize or bind tumor polypeptides. Such receptors include, but are not limited to, a fragment of the receptor, or a substitution, addition or deletion mutant of a T-cell receptor provided herein. This invention also encompasses polypeptides or peptides that are substantially homologous to the T-cell receptors provided herein or that retain substantially the same activity. The term “analog” includes any protein or polypeptide having an amino acid residue sequence substantially identical to the T-cell receptors provided herein in which one or more residues, preferably no more than 5 residues, more preferably no more than 25 residues have been conservatively substituted with a functionally similar residue and which displays the functional aspects of the T-cell receptor as described herein.  
       [0298] The present invention further provides for suitable mammalian host cells, for example, non-specific T cells, that are transfected with a polynucleotide encoding TCRs specific for a polypeptide described herein, thereby rendering the host cell specific for the polypeptide. The α and β chains of the TCR may be contained on separate expression vectors or alternatively, on a single expression vector that also contains an internal ribosome entry site (IRES) for cap-independent translation of the gene downstream of the IRES. Said host cells expressing TCRs specific for the polypeptide may be used, for example, for adoptive immunotherapy of lung cancer as discussed further below.  
       [0299] In further aspects of the present invention, cloned TCRs specific for a polypeptide recited herein may be used in a kit for the diagnosis of lung cancer. For example, the nucleic acid sequence or portions thereof, of tumor-specific TCRs can be used as probes or primers for the detection of expression of the rearranged genes encoding the specific TCR in a biological sample. Therefore, the present invention further provides for an assay for detecting messenger RNA or DNA encoding the TCR specific for a polypeptide.  
       [0300] Pharmaceutical Compositions  
       [0301] In additional embodiments, the present invention concerns formulation of one or more of the polynucleotide, polypeptide, T-cell, TCR, and/or antibody compositions disclosed herein in pharmaceutically-acceptable carriers for administration to a cell or an animal, either alone, or in combination with one or more other modalities of therapy.  
       [0302] It will be understood that, if desired, a composition as disclosed herein may be administered in combination with other agents as well, such as, e.g., other proteins or polypeptides or various pharmaceutically-active agents. In fact, there is virtually no limit to other components that may also be included, given that the additional agents do not cause a significant adverse effect upon contact with the target cells or host tissues. The compositions may thus be delivered along with various other agents as required in the particular instance. Such compositions may be purified from host cells or other biological sources, or alternatively may be chemically synthesized as described herein. Likewise, such compositions may further comprise substituted or derivatized RNA or DNA compositions.  
       [0303] Therefore, in another aspect of the present invention, pharmaceutical compositions are provided comprising one or more of the polynucleotide, polypeptide, antibody, TCR, and/or T-cell compositions described herein in combination with a physiologically acceptable carrier. In certain preferred embodiments, the pharmaceutical compositions of the invention comprise immunogenic polynucleotide and/or polypeptide compositions of the invention for use in prophylactic and theraputic vaccine applications. Vaccine preparation is generally described in, for example, M. F. Powell and M. J. Newman, eds., “Vaccine Design (the subunit and adjuvant approach),” Plenum Press (NY, 1995). Generally, such compositions will comprise one or more polynucleotide and/or polypeptide compositions of the present invention in combination with one or more immunostimulants.  
       [0304] It will be apparent that any of the pharmaceutical compositions described herein can contain pharmaceutically acceptable salts of the polynucleotides and polypeptides of the invention. Such salts can be prepared, for example, from pharmaceutically acceptable non-toxic bases, including organic bases (e.g., salts of primary, secondary and tertiary amines and basic amino acids) and inorganic bases (e.g., sodium, potassium, lithium, ammonium, calcium and magnesium salts).  
       [0305] In another embodiment, illustrative immunogenic compositions, e.g., vaccine compositions, of the present invention comprise DNA encoding one or more of the polypeptides as described above, such that the polypeptide is generated in situ. As noted above, the polynucleotide may be administered within any of a variety of delivery systems known to those of ordinary skill in the art. Indeed, numerous gene delivery techniques are well known in the art, such as those described by Rolland,  Crit. Rev. Therap. Drug Carrier Systems  15:143-198, 1998, and references cited therein. Appropriate polynucleotide expression systems will, of course, contain the necessary regulatory DNA regulatory sequences for expression in a patient (such as a suitable promoter and terminating signal). Alternatively, bacterial delivery systems may involve the administration of a bacterium (such as Bacillus-Calmette-Guerrin) that expresses an immunogenic portion of the polypeptide on its cell surface or secretes such an epitope.  
       [0306] Therefore, in certain embodiments, polynucleotides encoding immunogenic polypeptides described herein are introduced into suitable mammalian host cells for expression using any of a number of known viral-based systems. In one illustrative embodiment, retroviruses provide a convenient and effective platform for gene delivery systems. A selected nucleotide sequence encoding a polypeptide of the present invention can be inserted into a vector and packaged in retroviral particles using techniques known in the art. The recombinant virus can then be isolated and delivered to a subject. A number of illustrative retroviral systems have been described (e.g., U.S. Pat. No. 5,219,740; Miller and Rosman (1989) BioTechniques 7:980-990; Miller, A. D. (1990) Human Gene Therapy 1:5-14; Scarpa et al. (1991) Virology 180:849-852; Bums et al. (1993) Proc. Natl. Acad. Sci. USA 90:8033-8037; and Boris-Lawrie and Temin (1993) Cur. Opin. Genet. Develop. 3:102-109.  
       [0307] In addition, a number of illustrative adenovirus-based systems have also been described. Unlike retroviruses which integrate into the host genome, adenoviruses persist extrachromosomally thus minimizing the risks associated with insertional mutagenesis (Haj-Ahmad and Graham (1986) J. Virol. 57:267-274; Bett et al. (1993) J. Virol. 67:5911-5921; Mittereder et al. (1994) Human Gene Therapy 5:717-729; Seth et al. (1994) J. Virol. 68:933-940; Barr et al. (1994) Gene Therapy 1:51-58; Berkner, K. L. (1988) BioTechniques 6:616-629; and Rich et al. (1993) Human Gene Therapy 4:461-476).  
       [0308] Various adeno-associated virus (AAV) vector systems have also been developed for polynucleotide delivery. AAV vectors can be readily constructed using techniques well known in the art. See, e.g., U.S. Pat. Nos. 5,173,414 and 5,139,941; International Publication Nos. WO 92/01070 and WO 93/03769; Lebkowski et al. (1988) Molec. Cell. Biol. 8:3988-3996; Vincent et al. (1990) Vaccines 90 (Cold Spring Harbor Laboratory Press); Carter, B. J. (1992) Current Opinion in Biotechnology 3:533-539; Muzyczka, N. (1992) Current Topics in Microbiol. and Immunol. 158:97-129; Kotin, R. M. (1994) Human Gene Therapy 5:793-801; Shelling and Smith (1994) Gene Therapy 1:165-169; and Zhou et al. (1994) J. Exp. Med. 179:1867-1875.  
       [0309] Additional viral vectors useful for delivering the polynucleotides encoding polypeptides of the present invention by gene transfer include those derived from the pox family of viruses, such as vaccinia virus and avian poxvirus. By way of example, vaccinia virus recombinants expressing the novel molecules can be constructed as follows. The DNA encoding a polypeptide is first inserted into an appropriate vector so that it is adjacent to a vaccinia promoter and flanking vaccinia DNA sequences, such as the sequence encoding thymidine kinase (TK). This vector is then used to transfect cells which are simultaneously infected with vaccinia. Homologous recombination serves to insert the vaccinia promoter plus the gene encoding the polypeptide of interest into the viral genome. The resulting TK.sup.(−) recombinant can be selected by culturing the cells in the presence of 5-bromodeoxyuridine and picking viral plaques resistant thereto.  
       [0310] A vaccinia-based infection/transfection system can be conveniently used to provide for inducible, transient expression or coexpression of one or more polypeptides described herein in host cells of an organism. In this particular system, cells are first infected in vitro with a vaccinia virus recombinant that encodes the bacteriophage T7 RNA polymerase. This polymerase displays exquisite specificity in that it only transcribes templates bearing T7 promoters. Following infection, cells are transfected with the polynucleotide or polynucleotides of interest, driven by a T7 promoter. The polymerase expressed in the cytoplasm from the vaccinia virus recombinant transcribes the transfected DNA into RNA which is then translated into polypeptide by the host translational machinery. The method provides for high level, transient, cytoplasmic production of large quantities of RNA and its translation products. See, e.g., Elroy-Stein and Moss, Proc. Natl. Acad. Sci. USA (1990) 87:6743-6747; Fuerst et al. Proc. Natl. Acad. Sci. USA (1986) 83:8122-8126.  
       [0311] Alternatively, avipoxviruses, such as the fowlpox and canarypox viruses, can also be used to deliver the coding sequences of interest. Recombinant avipox viruses, expressing immunogens from mammalian pathogens, are known to confer protective immunity when administered to non-avian species. The use of an Avipox vector is particularly desirable in human and other mammalian species since members of the Avipox genus can only productively replicate in susceptible avian species and therefore are not infective in mammalian cells. Methods for producing recombinant Avipoxviruses are known in the art and employ genetic recombination, as described above with respect to the production of vaccinia viruses. See, e.g., WO 91/12882; WO 89/03429; and WO 92/03545.  
       [0312] Any of a number of alphavirus vectors can also be used for delivery of polynucleotide compositions of the present invention, such as those vectors described in U.S. Pat. Nos. 5,843,723; 6,015,686; 6,008,035 and 6,015,694. Certain vectors based on Venezuelan Equine Encephalitis (VEE) can also be used, illustrative examples of which can be found in U.S. Pat. Nos. 5,505,947 and 5,643,576.  
       [0313] Moreover, molecular conjugate vectors, such as the adenovirus chimeric vectors described in Michael et al. J. Biol. Chem. (1993) 268:6866-6869 and Wagner et al. Proc. Natl. Acad. Sci. USA (1992) 89:6099-6103, can also be used for gene delivery under the invention.  
       [0314] Additional illustrative information on these and other known viral-based delivery systems can be found, for example, in Fisher-Hoch et al.,  Proc. Natl. Acad. Sci. USA  86:317-321, 1989; Flexner et al.,  Ann. N.Y. Acad. Sci.  569:86-103, 1989; Flexner et al.,  Vaccine  8:17-21, 1990; U.S. Pat. Nos. 4,603,112, 4,769,330, and 5,017,487; WO 89/01973; U.S. Pat. No. 4,777,127; GB 2,200,651; EP 0,345,242; WO 91/02805; Berkner,  Biotechniques  6:616-627, 1988; Rosenfeld et al.,  Science  252:431-434, 1991; Kolls et al.,  Proc. Natl. Acad. Sci. USA  91:215-219, 1994; Kass-Eisler et al.,  Proc. Natl. Acad. Sci. USA  90:11498-11502, 1993; Guzman et al.,  Circulation  88:2838-2848, 1993; and Guzman et al.,  Cir. Res.  73:1202-1207, 1993.  
       [0315] In certain embodiments, a polynucleotide may be integrated into the genome of a target cell. This integration may be in the specific location and orientation via homologous recombination (gene replacement) or it may be integrated in a random, non-specific location (gene augmentation). In yet further embodiments, the polynucleotide may be stably maintained in the cell as a separate, episomal segment of DNA. Such polynucleotide segments or “episomes” encode sequences sufficient to permit maintenance and replication independent of or in synchronization with the host cell cycle. The manner in which the expression construct is delivered to a cell and where in the cell the polynucleotide remains is dependent on the type of expression construct employed.  
       [0316] In another embodiment of the invention, a polynucleotide is administered/delivered as “naked” DNA, for example as described in Ulmer et al.,  Science  259:1745-1749, 1993 and reviewed by Cohen,  Science  259:1691-1692, 1993. The uptake of naked DNA may be increased by coating the DNA onto biodegradable beads, which are efficiently transported into the cells.  
       [0317] In still another embodiment, a composition of the present invention can be delivered via a particle bombardment approach, many of which have been described. In one illustrative example, gas-driven particle acceleration can be achieved with devices such as those manufactured by Powderject Pharmaceuticals PLC (Oxford, UK) and Powderject Vaccines Inc. (Madison, Wis.), some examples of which are described in U.S. Pat. Nos. 5,846,796; 6,010,478; 5,865,796; 5,584,807; and EP Patent No. 0500 799. This approach offers a needle-free delivery approach wherein a dry powder formulation of microscopic particles, such as polynucleotide or polypeptide particles, are accelerated to high speed within a helium gas jet generated by a hand held device, propelling the particles into a target tissue of interest.  
       [0318] In a related embodiment, other devices and methods that may be useful for gas-driven needle-less injection of compositions of the present invention include those provided by Bioject, Inc. (Portland, Oreg.), some examples of which are described in U.S. Pat. Nos. 4,790,824; 5,064,413; 5,312,335; 5,383,851; 5,399,163; 5,520,639 and 5,993,412.  
       [0319] According to another embodiment, the pharmaceutical compositions described herein will comprise one or more immunostimulants in addition to the immunogenic polynucleotide, polypeptide, antibody, T-cell, TCR, and/or APC compositions of this invention. An immunostimulant refers to essentially any substance that enhances or potentiates an immune response (antibody and/or cell-mediated) to an exogenous antigen. One preferred type of immunostimulant comprises an adjuvant. Many adjuvants contain a substance designed to protect the antigen from rapid catabolism, such as aluminum hydroxide or mineral oil, and a stimulator of immune responses, such as lipid A,  Bortadella pertussis  or  Mycobacterium tuberculosis  derived proteins. Certain adjuvants are commercially available as, for example, Freund&#39;s Incomplete Adjuvant and Complete Adjuvant (Difco Laboratories, Detroit, Mich.); Merck Adjuvant 65 (Merck and Company, Inc., Rahway, N.J.); AS-2 (SmithKline Beecham, Philadelphia, Pa.); aluminum salts such as aluminum hydroxide gel (alum) or aluminum phosphate; salts of calcium, iron or zinc; an insoluble suspension of acylated tyrosine; acylated sugars; cationically or anionically derivatized polysaccharides; polyphosphazenes; biodegradable microspheres; monophosphoryl lipid A and quil A. Cytokines, such as GM-CSF, interleukin-2,-7,-12, and other like growth factors, may also be used as adjuvants.  
       [0320] Within certain embodiments of the invention, the adjuvant composition is preferably one that induces an immune response predominantly of the Th1 type. High levels of Th1-type cytokines (e.g., IFN-γ, TNFα, IL-2 and IL-12) tend to favor the induction of cell mediated immune responses to an administered antigen. In contrast, high levels of Th2-type cytokines (e.g., IL-4, IL-5, IL-6 and IL-10) tend to favor the induction of humoral immune responses. Following application of a vaccine as provided herein, a patient will support an immune response that includes Th1- and Th2-type responses. Within a preferred embodiment, in which a response is predominantly Th1-type, the level of Th1-type cytokines will increase to a greater extent than the level of Th2-type cytokines. The levels of these cytokines may be readily assessed using standard assays. For a review of the families of cytokines, see Mosmann and Coffinan,  Ann. Rev. Immunol.  7:145-173, 1989.  
       [0321] Certain preferred adjuvants for eliciting a predominantly Th1-type response include, for example, a combination of monophosphoryl lipid A, preferably 3-de-O-acylated monophosphoryl lipid A, together with an aluminum salt. MPL® adjuvants are available from Corixa Corporation (Seattle, Wash.; see, for example, U.S. Pat. Nos. 4,436,727; 4,877,611; 4,866,034 and 4,912,094). CpG-containing oligonucleotides (in which the CpG dinucleotide is unmethylated) also induce a predominantly Th1 response. Such oligonucleotides are well known and are described, for example, in WO 96/02555, WO 99/33488 and U.S. Pat. Nos. 6,008,200 and 5,856,462. Immunostimulatory DNA sequences are also described, for example, by Sato et al.,  Science  273:352, 1996. Another preferred adjuvant comprises a saponin, such as Quil A, or derivatives thereof, including QS21 and QS7 (Aquila Biopharmaceuticals Inc., Framingham, Mass.); Escin; Digitonin; or  Gypsophila  or  Chenopodium quinoa  saponins . Other preferred formulations include more than one saponin in the adjuvant combinations of the present invention, for example combinations of at least two of the following group comprising QS21, QS7, Quil A, β-escin, or digitonin.  
       [0322] Alternatively the saponin formulations may be combined with vaccine vehicles composed of chitosan or other polycationic polymers, polylactide and polylactide-co-glycolide particles, poly-N-acetyl glucosamine-based polymer matrix, particles composed of polysaccharides or chemically modified polysaccharides, liposomes and lipid-based particles, particles composed of glycerol monoesters, etc. The saponins may also be formulated in the presence of cholesterol to form particulate structures such as liposomes or ISCOMs. Furthermore, the saponins may be formulated together with a polyoxyethylene ether or ester, in either a non-particulate solution or suspension, or in a particulate structure such as a paucilamelar liposome or ISCOM. The saponins may also be formulated with excipients such as Carbopol R  to increase viscosity, or may be formulated in a dry powder form with a powder excipient such as lactose.  
       [0323] In one preferred embodiment, the adjuvant system includes the combination of a monophosphoryl lipid A and a saponin derivative, such as the combination of QS21 and 3D-MPL® adjuvant, as described in WO 94/00153, or a less reactogenic composition where the QS21 is quenched with cholesterol, as described in WO 96/33739. Other preferred formulations comprise an oil-in-water emulsion and tocopherol. Another particularly preferred adjuvant formulation employing QS21, 3D-MPL® adjuvant and tocopherol in an oil-in-water emulsion is described in WO 95/17210.  
       [0324] Another enhanced adjuvant system involves the combination of a CpG-containing oligonucleotide and a saponin derivative particularly the combination of CpG and QS21 is disclosed in WO 00/09159. Preferably the formulation additionally comprises an oil in water emulsion and tocopherol.  
       [0325] Additional illustrative adjuvants for use in the pharmaceutical compositions of the invention include Montanide ISA 720 (Seppic, France), SAF (Chiron, Calif., United States), ISCOMS (CSL), MF-59 (Chiron), the SBAS series of adjuvants (e.g., SBAS-2 or SBAS-4, available from SmithKline Beecham, Rixensart, Belgium), Detox (Enhanzyn®) (Corixa, Hamilton, Mont.), RC-529 (Corixa, Hamilton, Mont.) and other aminoalkyl glucosaminide 4-phosphates (AGPs), such as those described in pending U.S. patent application Ser. Nos. 08/853,826 and 09/074,720, the disclosures of which are incorporated herein by reference in their entireties, and polyoxyethylene ether adjuvants such as those described in WO 99/52549A1.  
       [0326] Other preferred adjuvants include adjuvant molecules of the general formula  
       (I): HO(CH 2 CH 2 O) n —A—R,  
       [0327] wherein, n is 1-50, A is a bond or —C(O)—, R is C 1-50  alkyl or Phenyl C 1-50  alkyl.  
       [0328] One embodiment of the present invention consists of a vaccine formulation comprising a polyoxyethylene ether of general formula (I), wherein n is between 1 and 50, preferably 4-24, most preferably 9; the R component is C 1-50 , preferably C 4 -C 20  alkyl and most preferably C 12  alkyl, and A is a bond. The concentration of the polyoxyethylene ethers should be in the range 0.1-20%, preferably from 0.1-10%, and most preferably in the range 0.1-1%. Preferred polyoxyethylene ethers are selected from the following group: polyoxyethylene-9-lauryl ether, polyoxyethylene-9-steoryl ether, polyoxyethylene-8-steoryl ether, polyoxyethylene-4-lauryl ether, polyoxyethylene-35-lauryl ether, and polyoxyethylene-23-lauryl ether. Polyoxyethylene ethers such as polyoxyethylene lauryl ether are described in the Merck index (12 th  edition: entry 7717). These adjuvant molecules are described in WO 99/52549.  
       [0329] The polyoxyethylene ether according to the general formula (I) above may, if desired, be combined with another adjuvant. For example, a preferred adjuvant combination is preferably with CpG as described in the pending UK patent application GB 9820956.2.  
       [0330] According to another embodiment of this invention, an immunogenic composition described herein is delivered to a host via antigen presenting cells (APCs), such as dendritic cells, macrophages, B cells, monocytes and other cells that may be engineered to be efficient APCs. Such cells may, but need not, be genetically modified to increase the capacity for presenting the antigen, to improve activation and/or maintenance of the T cell response, to have anti-tumor effects per se and/or to be immunologically compatible with the receiver (i.e., matched HLA haplotype). APCs may generally be isolated from any of a variety of biological fluids and organs, including tumor and peritumoral tissues, and may be autologous, allogeneic, syngeneic or xenogeneic cells.  
       [0331] Certain preferred embodiments of the present invention use dendritic cells or progenitors thereof as antigen-presenting cells. Dendritic cells are highly potent APCs (Banchereau and Steinman,  Nature  392:245-251, 1998) and have been shown to be effective as a physiological adjuvant for eliciting prophylactic or therapeutic antitumor immunity (see Timmerman and Levy,  Ann. Rev. Med.  50:507-529, 1999). In general, dendritic cells may be identified based on their typical shape (stellate in situ, with marked cytoplasmic processes (dendrites) visible in vitro), their ability to take up, process and present antigens with high efficiency and their ability to activate naïve T cell responses. Dendritic cells may, of course, be engineered to express specific cell-surface receptors or ligands that are not commonly found on dendritic cells in vivo or ex vivo, and such modified dendritic cells are contemplated by the present invention. As an alternative to dendritic cells, secreted vesicles antigen-loaded dendritic cells (called exosomes) may be used within a vaccine (see Zitvogel et al.,  Nature Med.  4:594-600, 1998).  
       [0332] Dendritic cells and progenitors may be obtained from peripheral blood, bone marrow, tumor-infiltrating cells, peritumoral tissues-infiltrating cells, lymph nodes, spleen, skin, umbilical cord blood or any other suitable tissue or fluid. For example, dendritic cells may be differentiated ex vivo by adding a combination of cytokines such as GM-CSF, IL-4, IL-13 and/or TNFα to cultures of monocytes harvested from peripheral blood. Alternatively, CD34 positive cells harvested from peripheral blood, umbilical cord blood or bone marrow may be differentiated into dendritic cells by adding to the culture medium combinations of GM-CSF, IL-3, TNFα, CD40 ligand, LPS, flt3 ligand and/or other compound(s) that induce differentiation, maturation and proliferation of dendritic cells.  
       [0333] Dendritic cells are conveniently categorized as “immature” and “mature” cells, which allows a simple way to discriminate between two well characterized phenotypes. However, this nomenclature should not be construed to exclude all possible intermediate stages of differentiation. Immature dendritic cells are characterized as APC with a high capacity for antigen uptake and processing, which correlates with the high expression of Fcγ receptor and mannose receptor. The mature phenotype is typically characterized by a lower expression of these markers, but a high expression of cell surface molecules responsible for T cell activation such as class I and class II MHC, adhesion molecules (e.g., CD54 and CD11) and costimulatory molecules (e.g., CD40, CD80, CD86 and 4-1BB).  
       [0334] APCs may generally be transfected with a polynucleotide of the invention (or portion or other variant thereof) such that the encoded polypeptide, or an immunogenic portion thereof, is expressed on the cell surface. Such transfection may take place ex vivo, and a pharmaceutical composition comprising such transfected cells may then be used for therapeutic purposes, as described herein. Alternatively, a gene delivery vehicle that targets a dendritic or other antigen presenting cell may be administered to a patient, resulting in transfection that occurs in vivo. In vivo and ex vivo transfection of dendritic cells, for example, may generally be performed using any methods known in the art, such as those described in WO 97/24447, or the gene gun approach described by Mahvi et al.,  Immunology and cell Biology  75:456-460, 1997. Antigen loading of dendritic cells may be achieved by incubating dendritic cells or progenitor cells with the tumor polypeptide, DNA (naked or within a plasmid vector) or RNA; or with antigen-expressing recombinant bacterium or viruses (e.g., vaccinia, fowlpox, adenovirus or lentivirus vectors). Prior to loading, the polypeptide may be covalently conjugated to an immunological partner that provides T cell help (e.g., a carrier molecule). Alternatively, a dendritic cell may be pulsed with a non-conjugated immunological partner, separately or in the presence of the polypeptide.  
       [0335] While any suitable carrier known to those of ordinary skill in the art may be employed in the pharmaceutical compositions of this invention, the type of carrier will typically vary depending on the mode of administration. Compositions of the present invention may be formulated for any appropriate manner of administration, including for example, topical, oral, nasal, mucosal, intravenous, intracranial, intraperitoneal, subcutaneous and intramuscular administration.  
       [0336] Carriers for use within such pharmaceutical compositions are biocompatible, and may also be biodegradable. In certain embodiments, the formulation preferably provides a relatively constant level of active component release. In other embodiments, however, a more rapid rate of release immediately upon administration may be desired. The formulation of such compositions is well within the level of ordinary skill in the art using known techniques. Illustrative carriers useful in this regard include microparticles of poly(lactide-co-glycolide), polyacrylate, latex, starch, cellulose, dextran and the like. Other illustrative delayed-release carriers include supramolecular biovectors, which comprise a non-liquid hydrophilic core (e.g., a cross-linked polysaccharide or oligosaccharide) and, optionally, an external layer comprising an amphiphilic compound, such as a phospholipid (see e.g., U.S. Pat. No. 5,151,254 and PCT applications WO 94/20078, WO/94/23701 and WO 96/06638). The amount of active compound contained within a sustained release formulation depends upon the site of implantation, the rate and expected duration of release and the nature of the condition to be treated or prevented.  
       [0337] In another illustrative embodiment, biodegradable microspheres (e.g., polylactate polyglycolate) are employed as carriers for the compositions of this invention. Suitable biodegradable microspheres are disclosed, for example, in U.S. Pat. Nos. 4,897,268; 5,075,109; 5,928,647; 5,811,128; 5,820,883; 5,853,763; 5,814,344, 5,407,609 and 5,942,252. Modified hepatitis B core protein carrier systems. such as described in WO/99 40934, and references cited therein, will also be useful for many applications. Another illustrative carrier/delivery system employs a carrier comprising particulate-protein complexes, such as those described in U.S. Pat. No. 5,928,647, which are capable of inducing a class I-restricted cytotoxic T lymphocyte responses in a host.  
       [0338] In another illustrative embodiment, calcium phosphate core particles are employed as carriers, vaccine adjuvants, or as controlled release matrices for the compositions of this invention. Exemplary calcium phosphate particles are disclosed, for example, in published patent application No. WO/0046147.  
       [0339] The pharmaceutical compositions of the invention will often further comprise one or more buffers (e.g., neutral buffered saline or phosphate buffered saline), carbohydrates (e.g., glucose, mannose, sucrose or dextrans), mannitol, proteins, polypeptides or amino acids such as glycine, antioxidants, bacteriostats, chelating agents such as EDTA or glutathione, adjuvants (e.g., aluminum hydroxide), solutes that render the formulation isotonic, hypotonic or weakly hypertonic with the blood of a recipient, suspending agents, thickening agents and/or preservatives. Alternatively, compositions of the present invention may be formulated as a lyophilizate.  
       [0340] The pharmaceutical compositions described herein may be presented in unit-dose or multi-dose containers, such as sealed ampoules or vials. Such containers are typically sealed in such a way to preserve the sterility and stability of the formulation until use. In general, formulations may be stored as suspensions, solutions or emulsions in oily or aqueous vehicles. Alternatively, a pharmaceutical composition may be stored in a freeze-dried condition requiring only the addition of a sterile liquid carrier immediately prior to use.  
       [0341] The development of suitable dosing and treatment regimens for using the particular compositions described herein in a variety of treatment regimens, including e.g., oral, parenteral, intravenous, intranasal, and intramuscular administration and formulation, is well known in the art, some of which are briefly discussed below for general purposes of illustration.  
       [0342] In certain applications, the pharmaceutical compositions disclosed herein may be delivered via oral administration to an animal. As such, these compositions may be formulated with an inert diluent or with an assimilable edible carrier, or they may be enclosed in hard- or soft-shell gelatin capsule, or they may be compressed into tablets, or they may be incorporated directly with the food of the diet.  
       [0343] The active compounds may even be incorporated with excipients and used in the form of ingestible tablets, buccal tables, troches, capsules, elixirs, suspensions, syrups, wafers, and the like (see, for example, Mathiowitz et al., Nature 1997 Mar 27;386(6623):410-4; Hwang et al., Crit Rev Ther Drug Carrier Syst 1998;15(3):243-84; U.S. Pat. No. 5,641,515; U.S. Pat. No. 5,580,579 and U.S. Pat. No. 5,792,451). Tablets, troches, pills, capsules and the like may also contain any of a variety of additional components, for example, a binder, such as gum tragacanth, acacia, cornstarch, or gelatin; excipients, such as dicalcium phosphate; a disintegrating agent, such as corn starch, potato starch, alginic acid and the like; a lubricant, such as magnesium stearate; and a sweetening agent, such as sucrose, lactose or saccharin may be added or a flavoring agent, such as peppermint, oil of wintergreen, or cherry flavoring. When the dosage unit form is a capsule, it may contain, in addition to materials of the above type, a liquid carrier. Various other materials may be present as coatings or to otherwise modify the physical form of the dosage unit. For instance, tablets, pills, or capsules may be coated with shellac, sugar, or both. Of course, any material used in preparing any dosage unit form should be pharmaceutically pure and substantially non-toxic in the amounts employed. In addition, the active compounds may be incorporated into sustained-release preparation and formulations.  
       [0344] Typically, these formulations will contain at least about 0.1% of the active compound or more, although the percentage of the active ingredient(s) may, of course, be varied and may conveniently be between about 1 or 2% and about 60% or 70% or more of the weight or volume of the total formulation. Naturally, the amount of active compound(s) in each therapeutically useful composition may be prepared is such a way that a suitable dosage will be obtained in any given unit dose of the compound. Factors such as solubility, bioavailability, biological half-life, route of administration, product shelf life, as well as other pharmacological considerations will be contemplated by one skilled in the art of preparing such pharmaceutical formulations, and as such, a variety of dosages and treatment regimens may be desirable.  
       [0345] For oral administration the compositions of the present invention may alternatively be incorporated with one or more excipients in the form of a mouthwash, dentifrice, buccal tablet, oral spray, or sublingual orally-administered formulation. Alternatively, the active ingredient may be incorporated into an oral solution such as one containing sodium borate, glycerin and potassium bicarbonate, or dispersed in a dentifrice, or added in a therapeutically-effective amount to a composition that may include water, binders, abrasives, flavoring agents, foaming agents, and humectants. Alternatively the compositions may be fashioned into a tablet or solution form that may be placed under the tongue or otherwise dissolved in the mouth.  
       [0346] In certain circumstances it will be desirable to deliver the pharmaceutical compositions disclosed herein parenterally, intravenously, intramuscularly, or even intraperitoneally. Such approaches are well known to the skilled artisan, some of which are further described, for example, in U.S. Pat. No. 5,543,158; U.S. Pat. No. 5,641,515 and U.S. Pat. No. 5,399,363. In certain embodiments, solutions of the active compounds as free base or pharmacologically acceptable salts may be prepared in water suitably mixed with a surfactant, such as hydroxypropylcellulose. Dispersions may also be prepared in glycerol, liquid polyethylene glycols, and mixtures thereof and in oils. Under ordinary conditions of storage and use, these preparations generally will contain a preservative to prevent the growth of microorganisms.  
       [0347] Illustrative pharmaceutical forms suitable for injectable use include sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions (for example, see U.S. Pat. No. 5,466,468). In all cases the form must be sterile and must be fluid to the extent that easy syringability exists. It must be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms, such as bacteria and fungi. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (e.g., glycerol, propylene glycol, and liquid polyethylene glycol, and the like), suitable mixtures thereof, and/or vegetable oils. Proper fluidity may be maintained, for example, by the use of a coating, such as lecithin, by the maintenance of the required particle size in the case of dispersion and/or by the use of surfactants. The prevention of the action of microorganisms can be facilitated by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like. In many cases, it will be preferable to include isotonic agents, for example, sugars or sodium chloride. Prolonged absorption of the injectable compositions can be brought about by the use in the compositions of agents delaying absorption, for example, aluminum monostearate and gelatin.  
       [0348] In one embodiment, for parenteral administration in an aqueous solution, the solution should be suitably buffered if necessary and the liquid diluent first rendered isotonic with sufficient saline or glucose. These particular aqueous solutions are especially suitable for intravenous, intramuscular, subcutaneous and intraperitoneal administration. In this connection, a sterile aqueous medium that can be employed will be known to those of skill in the art in light of the present disclosure. For example, one dosage may be dissolved in 1 ml of isotonic NaCl solution and either added to 1000 ml of hypodermoclysis fluid or injected at the proposed site of infusion, (see for example, “Remington&#39;s Pharmaceutical Sciences” 15th Edition, pages 1035-1038 and 1570-1580). Some variation in dosage will necessarily occur depending on the condition of the subject being treated. Moreover, for human administration, preparations will of course preferably meet sterility, pyrogenicity, and the general safety and purity standards as required by FDA Office of Biologics standards.  
       [0349] In another embodiment of the invention, the compositions disclosed herein may be formulated in a neutral or salt form. Illustrative pharmaceutically-acceptable salts include the acid addition salts (formed with the free amino groups of the protein) and which are formed with inorganic acids such as, for example, hydrochloric or phosphoric acids, or such organic acids as acetic, oxalic, tartaric, mandelic, and the like. Salts formed with the free carboxyl groups can also be derived from inorganic bases such as, for example, sodium, potassium, ammonium, calcium, or ferric hydroxides, and such organic bases as isopropylamine, trimethylamine, histidine, procaine and the like. Upon formulation, solutions will be administered in a manner compatible with the dosage formulation and in such amount as is therapeutically effective.  
       [0350] The carriers can further comprise any and all solvents, dispersion media, vehicles, coatings, diluents, antibacterial and antifungal agents, isotonic and absorption delaying agents, buffers, carrier solutions, suspensions, colloids, and the like. The use of such media and agents for pharmaceutical active substances is well known in the art. Except insofar as any conventional media or agent is incompatible with the active ingredient, its use in the therapeutic compositions is contemplated. Supplementary active ingredients can also be incorporated into the compositions. The phrase “pharmaceutically-acceptable” refers to molecular entities and compositions that do not produce an allergic or similar untoward reaction when administered to a human.  
       [0351] In certain embodiments, the pharmaceutical compositions may be delivered by intranasal sprays, inhalation, and/or other aerosol delivery vehicles. Methods for delivering genes, nucleic acids, and peptide compositions directly to the lungs via nasal aerosol sprays has been described, e.g., in U.S. Pat. No. 5,756,353 and U.S. Pat. No. 5,804,212. Likewise, the delivery of drugs using intranasal microparticle resins (Takenaga et al., J Controlled Release 1998 Mar 2;52(1-2):81-7) and lysophosphatidyl-glycerol compounds (U.S. Pat. No. 5,725,871) are also well-known in the pharmaceutical arts. Likewise, illustrative transmucosal drug delivery in the form of a polytetrafluoroetheylene support matrix is described in U.S. Pat. No. 5,780,045.  
       [0352] In certain embodiments, liposomes, nanocapsules, microparticles, lipid particles, vesicles, and the like, are used for the introduction of the compositions of the present invention into suitable host cells/organisms. In particular, the compositions of the present invention may be formulated for delivery either encapsulated in a lipid particle, a liposome, a vesicle, a nanosphere, or a nanoparticle or the like. Alternatively, compositions of the present invention can be bound, either covalently or non-covalently, to the surface of such carrier vehicles.  
       [0353] The formation and use of liposome and liposome-like preparations as potential drug carriers is generally known to those of skill in the art (see for example, Lasic, Trends Biotechnol 1998 Jul;16(7):307-21; Takakura, Nippon Rinsho 1998 Mar;56(3):691-5; Chandran et al., Indian J Exp Biol. 1997 Aug;35(8):801-9; Margalit, Crit Rev Ther Drug Carrier Syst. 1995;12(2-3):233-61; U.S. Pat. No. 5,567,434; U.S. Pat. No. 5,552,157; U.S. Pat. No. 5,565,213; U.S. Pat. No. 5,738,868 and U.S. Pat. No. 5,795,587, each specifically incorporated herein by reference in its entirety).  
       [0354] Liposomes have been used successfully with a number of cell types that are normally difficult to transfect by other procedures, including T cell suspensions, primary hepatocyte cultures and PC 12 cells (Renneisen et al., J Biol Chem. 1990 Sep 25;265(27):16337-42; Muller et al., DNA Cell Biol. 1990 Apr;9(3):221-9). In addition, liposomes are free of the DNA length constraints that are typical of viral-based delivery systems. Liposomes have been used effectively to introduce genes, various drugs, radiotherapeutic agents, enzymes, viruses, transcription factors, allosteric effectors and the like, into a variety of cultured cell lines and animals. Furthermore, he use of liposomes does not appear to be associated with autoimmune responses or unacceptable toxicity after systemic delivery.  
       [0355] In certain embodiments, liposomes are formed from phospholipids that are dispersed in an aqueous medium and spontaneously form multilamellar concentric bilayer vesicles (also termed multilamellar vesicles (MLVs).  
       [0356] Alternatively, in other embodiments, the invention provides for pharmaceutically-acceptable nanocapsule formulations of the compositions of the present invention. Nanocapsules can generally entrap compounds in a stable and reproducible way (see, for example, Quintanar-Guerrero et al., Drug Dev Ind Pharm. 1998 Dec;24(12):1113-28). To avoid side effects due to intracellular polymeric overloading, such ultrafine particles (sized around 0.1 μm) may be designed using polymers able to be degraded in vivo. Such particles can be made as described, for example, by Couvreur et al., Crit Rev Ther Drug Carrier Syst. 1988;5(1):1-20; zur Muhlen et al., Eur J Pharm Biopharm. 1998 Mar;45(2):149-55; Zambaux et al. J Controlled Release. 1998 Jan 2;50(1-3):31-40; and U.S. Pat. No. 5,145,684.  
       [0357] Cancer Therapeutic Methods  
       [0358] Immunologic approaches to cancer therapy are based on the recognition that cancer cells can often evade the body&#39;s defenses against aberrant or foreign cells and molecules, and that these defenses might be therapeutically stimulated to regain the lost ground, e.g. pgs. 623-648 in Klein, Immunology (Wiley-Interscience, New York, 1982). Numerous recent observations that various immune effectors can directly or indirectly inhibit growth of tumors has led to renewed interest in this approach to cancer therapy, e.g. Jager, et al., Oncology 2001;60(1):1-7; Renner, et al., Ann Hematol 2000 Dec;79(12):651-9.  
       [0359] Four-basic cell types whose function has been associated with antitumor cell immunity and the elimination of tumor cells from the body are: i) B-lymphocytes which secrete immunoglobulins into the blood plasma for identifying and labeling the nonself invader cells; ii) monocytes which secrete the complement proteins that are responsible for lysing and processing the immunoglobulin-coated target invader cells; iii) natural killer lymphocytes having two mechanisms for the destruction of tumor cells, antibody-dependent cellular cytotoxicity and natural killing; and iv) T-lymphocytes possessing antigen-specific receptors and having the capacity to recognize a tumor cell carrying complementary marker molecules (Schreiber, H., 1989, in Fundamental Immunology (ed). W. E. Paul, pp. 923-955).  
       [0360] Cancer immunotherapy generally focuses on inducing humoral immune responses, cellular immune responses, or both. Moreover, it is well established that induction of CD4 +  T helper cells is necessary in order to secondarily induce either antibodies or cytotoxic CD8 +  T cells. Polypeptide antigens that are selective or ideally specific for cancer cells, particularly lung cancer cells, offer a powerful approach for inducing immune responses against lung cancer, and are an important aspect of the present invention.  
       [0361] Therefore, in further aspects of the present invention, the pharmaceutical compositions described herein may be used to stimulate an immune response against cancer, particularly for the immunotherapy of lung cancer. Within such methods, the pharmaceutical compositions described herein are administered to a patient, typically a warm-blooded animal, preferably a human. A patient may or may not be afflicted with cancer. Pharmaceutical compositions and vaccines may be administered either prior to or following surgical removal of primary tumors and/or treatment such as administration of radiotherapy or conventional chemotherapeutic drugs. As discussed above, administration of the pharmaceutical compositions may be by any suitable method, including administration by intravenous, intraperitoneal, intramuscular, subcutaneous, intranasal, intradermal, anal, vaginal, topical and oral routes.  
       [0362] Within certain embodiments, immunotherapy may be active immunotherapy, in which treatment relies on the in vivo stimulation of the endogenous host immune system to react against tumors with the administration of immune response-modifying agents (such as polypeptides and polynucleotides as provided herein).  
       [0363] Within other embodiments, immunotherapy may be passive immunotherapy, in which treatment involves the delivery of agents with established tumor-immune reactivity (such as effector cells or antibodies) that can directly or indirectly mediate antitumor effects and does not necessarily depend on an intact host immune system. Examples of effector cells include T cells as discussed above, T lymphocytes (such as CD8 +  cytotoxic T lymphocytes and CD4 +  T-helper tumor-infiltrating lymphocytes), killer cells (such as Natural Killer cells and lymphokine-activated killer cells), B cells and antigen-presenting cells (such as dendritic cells and macrophages) expressing a polypeptide provided herein. T cell receptors and antibody receptors specific for the polypeptides recited herein may be cloned, expressed and transferred into other vectors or effector cells for adoptive immunotherapy. The polypeptides provided herein may also be used to generate antibodies or anti-idiotypic antibodies (as described above and in U.S. Pat. No. 4,918,164) for passive immunotherapy.  
       [0364] Monoclonal antibodies may be labeled with any of a variety of labels for desired selective usages in detection, diagnostic assays or therapeutic applications (as described in U.S. Pat. Nos. 6,090,365; 6,015,542; 5,843,398; 5,595,721; and 4,708,930, hereby incorporated by reference in their entirety as if each was incorporated individually). In each case, the binding of the labelled monoclonal antibody to the determinant site of the antigen will signal detection or delivery of a particular therapeutic agent to the antigenic determinant on the non-normal cell. A further object of this invention is to provide the specific monoclonal antibody suitably labelled for achieving such desired selective usages thereof.  
       [0365] Effector cells may generally be obtained in sufficient quantities for adoptive immunotherapy by growth in vitro, as described herein. Culture conditions for expanding single antigen-specific effector cells to several billion in number with retention of antigen recognition in vivo are well known in the art. Such in vitro culture conditions typically use intermittent stimulation with antigen, often in the presence of cytokines (such as IL-2) and non-dividing feeder cells. As noted above, immunoreactive polypeptides as provided herein may be used to rapidly expand antigen-specific T cell cultures in order to generate a sufficient number of cells for immunotherapy. In particular, antigen-presenting cells, such as dendritic, macrophage, monocyte, fibroblast and/or B cells, may be pulsed with immunoreactive polypeptides or transfected with one or more polynucleotides using standard techniques well known in the art. For example, antigen-presenting cells can be transfected with a polynucleotide having a promoter appropriate for increasing expression in a recombinant virus or other expression system. Cultured effector cells for use in therapy must be able to grow and distribute widely, and to survive long term in vivo. Studies have shown that cultured effector cells can be induced to grow in vivo and to survive long term in substantial numbers by repeated stimulation with antigen supplemented with IL-2 (see, for example, Cheever et al.,  Immunological Reviews  157:177, 1997).  
       [0366] Alternatively, a vector expressing a polypeptide recited herein may be introduced into antigen presenting cells taken from a patient and clonally propagated ex vivo for transplant back into the same patient. Transfected cells may be reintroduced into the patient using any means known in the art, preferably in sterile form by intravenous, intracavitary, intraperitoneal or intratumor administration.  
       [0367] Routes and frequency of administration of the therapeutic compositions described herein, as well as dosage, will vary from individual to individual, and may be readily established using standard techniques. In general, the pharmaceutical compositions and vaccines may be administered by injection (e.g., intracutaneous, intramuscular, intravenous or subcutaneous), intranasally (e.g., by aspiration) or orally. Preferably, between 1 and 10 doses may be administered over a 52 week period. Preferably, 6 doses are administered, at intervals of 1 month, and booster vaccinations may be given periodically thereafter. Alternate protocols may be appropriate for individual patients. A suitable dose is an amount of a compound that, when administered as described above, is capable of promoting an anti-tumor immune response, and is at least 10-50% above the basal (i.e., untreated) level. Such response can be monitored by measuring the anti-tumor antibodies in a patient or by vaccine-dependent generation of cytolytic effector cells capable of killing the patient&#39;s tumor cells in vitro. Such vaccines should also be capable of causing an immune response that leads to an improved clinical outcome (e.g., more frequent remissions, complete or partial or longer disease-free survival) in vaccinated patients as compared to non-vaccinated patients. In general, for pharmaceutical compositions and vaccines comprising one or more polypeptides, the amount of each polypeptide present in a dose ranges from about 25 μg to 5 mg per kg of host. Suitable dose sizes will vary with the size of the patient, but will typically range from about 0.1 mL to about 5 mL.  
       [0368] In general, an appropriate dosage and treatment regimen provides the active compound(s) in an amount sufficient to provide therapeutic and/or prophylactic benefit. Such a response can be monitored by establishing an improved clinical outcome (e.g., more frequent remissions, complete or partial, or longer disease-free survival) in treated patients as compared to non-treated patients. Increases in preexisting immune responses to a tumor protein generally correlate with an improved clinical outcome. Such immune responses may generally be evaluated using standard proliferation, cytotoxicity or cytokine assays, which may be performed using samples obtained from a patient before and after treatment.  
       [0369] Cancer Detection and Diagnostic Compositions Methods and Kits  
       [0370] In general, a cancer may be detected in a patient based on the presence of one or more lung tumor proteins and/or polynucleotides encoding such proteins in a biological sample (for example, blood, sera, sputum urine and/or tumor biopsies) obtained from the patient. In other words, such proteins may be used as markers to indicate the presence or absence of a cancer such as lung cancer. In addition, such proteins may be useful for the detection of other cancers. The binding agents provided herein generally permit detection of the level of antigen that binds to the agent in the biological sample.  
       [0371] Polynucleotide primers and probes may be used to detect the level of mRNA encoding a tumor protein, which is also indicative of the presence or absence of a cancer. In general, a tumor sequence should be present at a level that is at least two-fold, preferably three-fold, and more preferably five-fold or higher in tumor tissue than in normal tissue of the same type from which the tumor arose. Expression levels of a particular tumor sequence in tissue types different from that in which the tumor arose are irrelevant in certain diagnostic embodiments since the presence of tumor cells can be confirmed by observation of predetermined differential expression levels, e.g., 2-fold, 5-fold, etc, in tumor tissue to expression levels in normal tissue of the same type.  
       [0372] Other differential expression patterns can be utilized advantageously for diagnostic purposes. For example, in one aspect of the invention, overexpression of a tumor sequence in tumor tissue and normal tissue of the same type, but not in other normal tissue types, e.g. PBMCs, can be exploited diagnostically. In this case, the presence of metastatic tumor cells, for example in a sample taken from the circulation or some other tissue site different from that in which the tumor arose, can be identified and/or confirmed by detecting expression of the tumor sequence in the sample, for example using RT-PCR analysis. In many instances, it will be desired to enrich for tumor cells in the sample of interest, e.g., PBMCs, using cell capture or other like techniques.  
       [0373] There are a variety of assay formats known to those of ordinary skill in the art for using a binding agent to detect polypeptide markers in a sample. See, e.g., Harlow and Lane,  Antibodies: A Laboratory Manual,  Cold Spring Harbor Laboratory, 1988. In general, the presence or absence of a cancer in a patient may be determined by (a) contacting a biological sample obtained from a patient with a binding agent; (b) detecting in the sample a level of polypeptide that binds to the binding agent; and (c) comparing the level of polypeptide with a predetermined cut-off value.  
       [0374] In a preferred embodiment, the assay involves the use of binding agent immobilized on a solid support to bind to and remove the polypeptide from the remainder of the sample. The bound polypeptide may then be detected using a detection reagent that contains a reporter group and specifically binds to the binding agent/polypeptide complex. Such detection reagents may comprise, for example, a binding agent that specifically binds to the polypeptide or an antibody or other agent that specifically binds to the binding agent, such as an anti-immunoglobulin, protein G, protein A or a lectin. Alternatively, a competitive assay may be utilized, in which a polypeptide is labeled with a reporter group and allowed to bind to the immobilized binding agent after incubation of the binding agent with the sample. The extent to which components of the sample inhibit the binding of the labeled polypeptide to the binding agent is indicative of the reactivity of the sample with the immobilized binding agent. Suitable polypeptides for use within such assays include full length lung tumor proteins and polypeptide portions thereof to which the binding agent binds, as described above.  
       [0375] The solid support may be any material known to those of ordinary skill in the art to which the tumor protein may be attached. For example, the solid support may be a test well in a microtiter plate or a nitrocellulose or other suitable membrane. Alternatively, the support may be a bead or disc, such as glass, fiberglass, latex or a plastic material such as polystyrene or polyvinylchloride. The support may also be a magnetic particle or a fiber optic sensor, such as those disclosed, for example, in U.S. Pat. No. 5,359,681. The binding agent may be immobilized on the solid support using a variety of techniques known to those of skill in the art, which are amply described in the patent and scientific literature. In the context of the present invention, the term “immobilization” refers to both noncovalent association, such as adsorption, and covalent attachment (which may be a direct linkage between the agent and functional groups on the support or may be a linkage by way of a cross-linking agent). Immobilization by adsorption to a well in a microtiter plate or to a membrane is preferred. In such cases, adsorption may be achieved by contacting the binding agent, in a suitable buffer, with the solid support for a suitable amount of time. The contact time varies with temperature, but is typically between about 1 hour and about 1 day. In general, contacting a well of a plastic microtiter plate (such as polystyrene or polyvinylchloride) with an amount of binding agent ranging from about 10 ng to about 10 μg, and preferably about 100 ng to about 1 μg, is sufficient to immobilize an adequate amount of binding agent.  
       [0376] Covalent attachment of binding agent to a solid support may generally be achieved by first reacting the support with a bifunctional reagent that will react with both the support and a functional group, such as a hydroxyl or amino group, on the binding agent. For example, the binding agent may be covalently attached to supports having an appropriate polymer coating using benzoquinone or by condensation of an aldehyde group on the support with an amine and an active hydrogen on the binding partner (see, e.g., Pierce Immunotechnology Catalog and Handbook, 1991, at A12-A13).  
       [0377] In certain embodiments, the assay is a two-antibody sandwich assay. This assay may be performed by first contacting an antibody that has been immobilized on a solid support, commonly the well of a microtiter plate, with the sample, such that polypeptides within the sample are allowed to bind to the immobilized antibody. Unbound sample is then removed from the immobilized polypeptide-antibody complexes and a detection reagent (preferably a second antibody capable of binding to a different site on the polypeptide) containing a reporter group is added. The amount of detection reagent that remains bound to the solid support is then determined using a method appropriate for the specific reporter group.  
       [0378] More specifically, once the antibody is immobilized on the support as described above, the remaining protein binding sites on the support are typically blocked. Any suitable blocking agent known to those of ordinary skill in the art, such as bovine serum albumin or Tween 20™ (Sigma Chemical Co., St. Louis, Mo.). The immobilized antibody is then incubated with the sample, and polypeptide is allowed to bind to the antibody. The sample may be diluted with a suitable diluent, such as phosphate-buffered saline (PBS) prior to incubation. In general, an appropriate contact time (i.e., incubation time) is a period of time that is sufficient to detect the presence of polypeptide within a sample obtained from an individual with lung least about 95% of that achieved at equilibrium between bound and unbound polypeptide. Those of ordinary skill in the art will recognize that the time necessary to achieve equilibrium may be readily determined by assaying the level of binding that occurs over a period of time. At room temperature, an incubation time of about 30 minutes is generally sufficient.  
       [0379] Unbound sample may then be removed by washing the solid support with an appropriate buffer, such as PBS containing 0.1% Tween 20™. The second antibody, which contains a reporter group, may then be added to the solid support. Preferred reporter groups include those groups recited above.  
       [0380] The detection reagent is then incubated with the immobilized antibody-polypeptide complex for an amount of time sufficient to detect the bound polypeptide. An appropriate amount of time may generally be determined by assaying the level of binding that occurs over a period of time. Unbound detection reagent is then removed and bound detection reagent is detected using the reporter group. The method employed for detecting the reporter group depends upon the nature of the reporter group. For radioactive groups, scintillation counting or autoradiographic methods are generally appropriate. Spectroscopic methods may be used to detect dyes, luminescent groups and fluorescent groups. Biotin may be detected using avidin, coupled to a different reporter group (commonly a radioactive or fluorescent group or an enzyme). Enzyme reporter groups may generally be detected by the addition of substrate (generally for a specific period of time), followed by spectroscopic or other analysis of the reaction products.  
       [0381] To determine the presence or absence of a cancer, such as lung cancer, the signal detected from the reporter group that remains bound to the solid support is generally compared to a signal that corresponds to a predetermined cut-off value. In one preferred embodiment, the cut-off value for the detection of a cancer is the average mean signal obtained when the immobilized antibody is incubated with samples from patients without the cancer. In general, a sample generating a signal that is three standard deviations above the predetermined cut-off value is considered positive for the cancer. In an alternate preferred embodiment, the cut-off value is determined using a Receiver Operator Curve, according to the method of Sackett et al.,  Clinical Epidemiology: A Basic Science for Clinical Medicine,  Little Brown and Co., 1985, p. 106-7. Briefly, in this embodiment, the cut-off value may be determined from a plot of pairs of true positive rates (i.e., sensitivity) and false positive rates (100%-specificity) that correspond to each possible cut-off value for the diagnostic test result. The cut-off value on the plot that is the closest to the upper left-hand corner (i.e., the value that encloses the largest area) is the most accurate cut-off value, and a sample generating a signal that is higher than the cut-off value determined by this method may be considered positive. Alternatively, the cut-off value may be shifted to the left along the plot, to minimize the false positive rate, or to the right, to minimize the false negative rate. In general, a sample generating a signal that is higher than the cut-off value determined by this method is considered positive for a cancer.  
       [0382] In a related embodiment, the assay is performed in a flow-through or strip test format, wherein the binding agent is immobilized on a membrane, such as nitrocellulose. In the flow-through test, polypeptides within the sample bind to the immobilized binding agent as the sample passes through the membrane. A second, labeled binding agent then binds to the binding agent-polypeptide complex as a solution containing the second binding agent flows through the membrane. The detection of bound second binding agent may then be performed as described above. In the strip test format, one end of the membrane to which binding agent is bound is immersed in a solution containing the sample. The sample migrates along the membrane through a region containing second binding agent and to the area of immobilized binding agent. Concentration of second binding agent at the area of immobilized antibody indicates the presence of a cancer. Typically, the concentration of second binding agent at that site generates a pattern, such as a line, that can be read visually. The absence of such a pattern indicates a negative result. In general, the amount of binding agent immobilized on the membrane is selected to generate a visually discernible pattern when the biological sample contains a level of polypeptide that would be sufficient to generate a positive signal in the two-antibody sandwich assay, in the format discussed above. Preferred binding agents for use in such assays are antibodies and antigen-binding fragments thereof. Preferably, the amount of antibody immobilized on the membrane ranges from about 25 ng to about 1 μg, and more preferably from about 50 ng to about 500 ng. Such tests can typically be performed with a very small amount of biological sample.  
       [0383] Of course, numerous other assay protocols exist that are suitable for use with the tumor proteins or binding agents of the present invention. The above descriptions are intended to be exemplary only. For example, it will be apparent to those of ordinary skill in the art that the above protocols may be readily modified to use tumor polypeptides to detect antibodies that bind to such polypeptides in a biological sample. The detection of such tumor protein specific antibodies may correlate with the presence of a cancer.  
       [0384] A cancer may also, or alternatively, be detected based on the presence of T cells that specifically react with a tumor protein in a biological sample. Within certain methods, a biological sample comprising CD4 +  and/or CD8 +  T cells isolated from a patient is incubated with a tumor polypeptide, a polynucleotide encoding such a polypeptide and/or an APC that expresses at least an immunogenic portion of such a polypeptide, and the presence or absence of specific activation of the T cells is detected. Suitable biological samples include, but are not limited to, isolated T cells. For example, T cells may be isolated from a patient by routine techniques (such as by Ficoll/Hypaque density gradient centrifugation of peripheral blood lymphocytes). T cells may be incubated in vitro for 2-9 days (typically 4 days) at 37° C. with polypeptide (e.g., 5-25 μg/ml). It may be desirable to incubate another aliquot of a T cell sample in the absence of tumor polypeptide to serve as a control. For CD4 +  T cells, activation is preferably detected by evaluating proliferation of the T cells. For CD8 +  T cells, activation is preferably detected by evaluating cytolytic activity. A level of proliferation that is at least two fold greater and/or a level of cytolytic activity that is at least 20% greater than in disease-free patients indicates the presence of a cancer in the patient.  
       [0385] As noted above, a cancer may also, or alternatively, be detected based on the level of mRNA encoding a tumor protein in a biological sample. For example, at least two oligonucleotide primers may be employed in a polymerase chain reaction (PCR) based assay to amplify a portion of a tumor cDNA derived from a biological sample, wherein at least one of the oligonucleotide primers is specific for (i.e., hybridizes to) a polynucleotide encoding the tumor protein. The amplified cDNA is then separated and detected using techniques well known in the art, such as gel electrophoresis.  
       [0386] Similarly, oligonucleotide probes that specifically hybridize to a polynucleotide encoding a tumor protein may be used in a hybridization assay to detect the presence of polynucleotide encoding the tumor protein in a biological sample.  
       [0387] To permit hybridization under assay conditions, oligonucleotide primers and probes should comprise an oligonucleotide sequence that has at least about 60%, preferably at least about 75% and more preferably at least about 90%, identity to a portion of a polynucleotide encoding a tumor protein of the invention that is at least 10 nucleotides, and preferably at least 20 nucleotides, in length. Preferably, oligonucleotide primers and/or probes hybridize to a polynucleotide encoding a polypeptide described herein under moderately stringent conditions, as defined above. Oligonucleotide primers and/or probes which may be usefully employed in the diagnostic methods described herein preferably are at least 10-40 nucleotides in length. In a preferred embodiment, the oligonucleotide primers comprise at least 10 contiguous nucleotides, more preferably at least 15 contiguous nucleotides, of a DNA molecule having a sequence as disclosed herein. Techniques for both PCR based assays and hybridization assays are well known in the art (see, for example, Mullis et al.,  Cold Spring Harbor Symp. Quant. Biol.,  51:263, 1987; Erlich ed., PCR Technology, Stockton Press, NY, 1989).  
       [0388] One preferred assay employs RT-PCR, in which PCR is applied in conjunction with reverse transcription. Typically, RNA is extracted from a biological sample, such as biopsy tissue, and is reverse transcribed to produce cDNA molecules. PCR amplification using at least one specific primer generates a cDNA molecule, which may be separated and visualized using, for example, gel electrophoresis. Amplification may be performed on biological samples taken from a test patient and from an individual who is not afflicted with a cancer. The amplification reaction may be performed on several dilutions of cDNA spanning two orders of magnitude. A two-fold or greater increase in expression in several dilutions of the test patient sample as compared to the same dilutions of the non-cancerous sample is typically considered positive.  
       [0389] In another aspect of the present invention, cell capture technologies may be used in conjunction, with, for example, real-time PCR to provide a more sensitive tool for detection of metastatic cells expressing lung tumor antigens. Detection of lung cancer cells in biological samples, e.g., bone marrow samples, peripheral blood, and small needle aspiration samples is desirable for diagnosis and prognosis in lung cancer patients.  
       [0390] Immunomagnetic beads coated with specific monoclonal antibodies to surface cell markers, or tetrameric antibody complexes, may be used to first enrich or positively select cancer cells in a sample. Various commercially available kits may be used, including Dynabeads® Epithelial Enrich (Dynal Biotech, Oslo, Norway), StemSep™ (StemCell Technologies, Inc., Vancouver, BC), and RosetteSep (StemCell Technologies). A skilled artisan will recognize that other methodologies and kits may also be used to enrich or positively select desired cell populations. Dynabeads® Epithelial Enrich contains magnetic beads coated with mAbs specific for two glycoprotein membrane antigens expressed on normal and neoplastic epithelial tissues. The coated beads may be added to a sample and the sample then applied to a magnet, thereby capturing the cells bound to the beads. The unwanted cells are washed away and the magnetically isolated cells eluted from the beads and used in further analyses.  
       [0391] RosetteSep can be used to enrich cells directly from a blood sample and consists of a cocktail of tetrameric antibodies that targets a variety of unwanted cells and crosslinks them to glycophorin A on red blood cells (RBC) present in the sample, forming rosettes. When centrifuged over Ficoll, targeted cells pellet along with the free RBC. The combination of antibodies in the depletion cocktail determines which cells will be removed and consequently which cells will be recovered. Antibodies that are available include, but are not limited to: CD2, CD3, CD4, CD5, CD8, CD10, CD11b, CD14, CD15, CD16, CD19, CD20, CD24, CD25, CD29, CD33, CD34, CD36, CD38, CD41, CD45, CD45RA, CD45RO, CD56, CD66B, CD66e, HLA-DR, IgE, and TCRαβ.  
       [0392] Additionally, it is contemplated in the present invention that mAbs specific for lung tumor antigens can be generated and used in a similar manner. For example, mAbs that bind to tumor-specific cell surface antigens may be conjugated to magnetic beads, or formulated in a tetrameric antibody complex, and used to enrich or positively select metastatic lung tumor cells from a sample. Once a sample is enriched or positively selected, cells may be lysed and RNA isolated. RNA may then be subjected to RT-PCR analysis using lung tumor-specific primers in a real-time PCR assay as described herein. One skilled in the art will recognize that enriched or selected populations of cells may be analyzed by other methods (e.g. in situ hybridization or flow cytometry).  
       [0393] In another embodiment, the compositions described herein may be used as markers for the progression of cancer. In this embodiment, assays as described above for the diagnosis of a cancer may be performed over time, and the change in the level of reactive polypeptide(s) or polynucleotide(s) evaluated. For example, the assays may be performed every 24-72 hours for a period of 6 months to 1 year, and thereafter performed as needed. In general, a cancer is progressing in those patients in whom the level of polypeptide or polynucleotide detected increases over time. In contrast, the cancer is not progressing when the level of reactive polypeptide or polynucleotide either remains constant or decreases with time.  
       [0394] Certain in vivo diagnostic assays may be performed directly on a tumor. One such assay involves contacting tumor cells with a binding agent. The bound binding agent may then be detected directly or indirectly via a reporter group. Such binding agents may also be used in histological applications. Alternatively, polynucleotide probes may be used within such applications.  
       [0395] As noted above, to improve sensitivity, multiple tumor protein markers may be assayed within a given sample. It will be apparent that binding agents specific for different proteins provided herein may be combined within a single assay. Further, multiple primers or probes may be used concurrently. The selection of tumor protein markers may be based on routine experiments to determine combinations that results in optimal sensitivity. In addition, or alternatively, assays for tumor proteins provided herein may be combined with assays for other known tumor antigens.  
       [0396] The present invention further provides kits for use within any of the above diagnostic methods. Such kits typically comprise two or more components necessary for performing a diagnostic assay. Components may be compounds, reagents, containers and/or equipment. For example, one container within a kit may contain a monoclonal antibody or fragment thereof that specifically binds to a tumor protein. Such antibodies or fragments may be provided attached to a support material, as described above. One or more additional containers may enclose elements, such as reagents or buffers, to be used in the assay. Such kits may also, or alternatively, contain a detection reagent as described above that contains a reporter group suitable for direct or indirect detection of antibody binding.  
       [0397] Alternatively, a kit may be designed to detect the level of mRNA encoding a tumor protein in a biological sample. Such kits generally comprise at least one oligonucleotide probe or primer, as described above, that hybridizes to a polynucleotide encoding a tumor protein. Such an oligonucleotide may be used, for example, within a PCR or hybridization assay. Additional components that may be present within such kits include a second oligonucleotide and/or a diagnostic reagent or container to facilitate the detection of a polynucleotide encoding a tumor protein.  
       [0398] The following Examples are offered by way of illustration and not by way of limitation. 
     
    
    
     EXAMPLES  
     Example 1  
     Identification of Lung Tumor Protein cDNAs  
     [0399] Lung-specific genes were identified by electronic subtraction. The method used was similar to that described by Vasmatizis et al.,  Proc. Natl. Acad. Sci. USA  95:300-304, 1998, but there were several key differences. Sequences of EST clones (1,453,679) were downloaded from the GenBank public human EST database. Human cDNA libraries were downloaded to create a database of these cDNA libraries and the EST sequences derived from them. The cDNA libraries were grouped into three groups: Plus, Minus and Other/Neutral. The Plus group included 30 libraries constructed from lung tumor and fetal lung tissues (and therefore including those containing lung tumor-specific ESTs); the Minus group consisted of 206 libraries derived from all adult normal tissues; the Other/Neutral group contained libraries from tissues where expression is considered irrelevant (e.g., non-lung-fetal tissue, non-lung tumors, cell lines other than lung tumor cell lines). A total of 93,526 ESTs were derived from the 30 lung tumor and fetal lung libraries. These ESTs were preprocessed to remove common sequence repeats and cloning adapters, resulting in a final Plus group of 90,365 (a decrease of 3%).  
     [0400] Each Plus group (lung tumor or fetal lung) EST sequence was used as a query “seed” sequence in a BLASTN (version 2.0.9; May 7, 1999) search against the total human EST database. Standard measures of similarity are insufficient in this sort of analysis, as EST relationships often include short stretches and poor sequence data. Criteria employed in this study required a matching segment to be at least 75 nucleotides in length, and the density of exact matches within this segment to be at least 80%. This was considered conservative criteria designed to avoid short spurious matches while allowing for polymorphisms and errors in sequencing. Each BLAST search generated a cluster of related sequences based on direct overlap with the query “seed” sequence. A second level of clustering was performed to merge closely related clusters and to eliminate redundancy resulting from the fact that similar clusters are generated if the clusters contain more than one seed (i.e., sequences from the Plus EST group). The resulting “super clusters” were discarded if they grew in size to 200 or more ESTs, since these probably represented repetitive elements that were not removed by the initial preprocessing of the seeds, or highly expressed genes such as those for ribosomal proteins. Superclusters were merged if they shared at least one third of their sequences.  
     [0401] The BLAST searches gave rise to a total of 49,154 clusters. In the first super clustering stage, 18,665 clusters grew beyond the limit of 200 clones. The remainder was reduced to a total of 30,489 super clusters. This number was reduced to 29,501 after adjacent clusters were merged. Resulting super clusters were analyzed to determine the tissue source of each EST clone contained within it and this expression profile was used to classify the superclusters into four groups: Type 1- this supercluster contains EST clones found in the Plus group only, with no expression in the Minus or Other/Neutral group libraries; Type 2—EST clones in the supercluster are found in the Plus and Other/Neutral group libraries, with no expression in the Minus group; Type 3—super cluster EST clones found in all groups, but the number of ESTs in the Plus group is higher than in either of the Minus or Other/Neutral groups; Type 4—super cluster EST clones found in all groups, but the number in the Plus group is higher than in the Minus group with expression in the Other/Neutral group non relevant. Sequences derived from the Plus library group that were placed in Types 1, 2 and 3 superclusters resulted in 20,487 polynucleotide sequences. The electronic subtraction procedures identified these sequences as having significant differential expression in lung tissue.  
     Example 2  
     Analysis of CDNA Expression Using Microarray Technology  
     [0402] 2208 of the clones identified from the lung electronic subtraction procedure were evaluated for overexpression in specific tumor tissues by microarray analysis. Using this approach, cDNA sequences are PCR amplified and their mRNA expression profiles in tumor and normal tissues are examined using cDNA microarray technology essentially as described (Shena, M. et al., 1995 Science 270:467-70). In brief, the 2208 clones were arrayed onto glass slides as multiple replicas, with each location corresponding to a unique cDNA clone (as many as 5500 clones can be arrayed on a single slide or chip). Each chip was hybridized with a pair of cDNA probes that were fluorescence-labeled with Cy3 and Cy5, respectively. Typically, 1 μg of polyA +  RNA was used to generate each cDNA probe. Since one cDNA probe is generated from tumor tissue RNA and the other is generated from normal tissue RNA, sequences that are differentially overexpressed in tumor tissue will generate a stronger signal from the tumor specific probe than the normal tissue probe, thus allowing the identification of those sequences that exhibit elevated expression in tumor versus normal tissue.  
     [0403] After hybridization, the chips were scanned and the fluorescence intensity recorded for both Cy3 and Cy5 channels. There were multiple built-in quality control steps. First, the probe quality was monitored using a panel of 18 ubiquitously expressed genes. Secondly, the control plate also had yeast DNA fragments of which complementary RNA was spiked into the probe synthesis for measuring the quality of the probe and the sensitivity of the analysis. Currently, the technology offers a sensitivity of 1 in 100,000 copies of mRNA. Finally, the reproducibility of this technology was ensured by including duplicated control cDNA elements at different locations. Further validation of the process was indicated in that several differentially expressed genes were identified multiple times in the study, and the expression profiles for these genes are very comparable. The clones were arrayed on Lung Chip 6.  
     [0404] Of those analyzed by microarray, 781 sequences met the criteria of having at least 2-fold overexpression in lung tumor tissue compared to normal tissues. Of these 781 clones, 459 were found to meet the additional criteria of having a mean normal tissue expression value less than or equal to 0.2. These 459 clones were then analyzed visually and certain ones with favorable expression profiles (e.g., high expression in tumors with little or no expression in normal tissues) were sequenced and searched against public sequences databases to facilitate identification of extended sequence for the clones.  
     [0405] SEQ ID NO:1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 32 and 34 represent a subset of those 459 clones that met the above criteria of being at least 2-fold overexpressed in tumor versus normal tissues and having a mean normal tissue expression of less than or equal to 0.2. Additional information about these sequences is provided in Table 2 below.  
                                   TABLE 2                                       MICRO-   MICRO-           SEQ ID           ARRAY   ARRAY RATIO       SEQ   NO:           ANALYSIS   (Lung       ID   from   Clone   Clone   (Lung   Tumor:Normal       NO:   60/207,485   Name:   ID #   Chip #)   Tissue)                                                        9   4538   L1027C   55571   6   2.94       5   4978   L1037C   58267   6   2.61       7   1796   L1038C   58245   6   3.5       3   7264   L1039C   58269   6   2.81       1   2337   L1040C   55964   6   5.07       15   1548/4619   L1041C   58346   6   2.33       25   15127    n/a   56016   6   &gt;2       27   3816   n/a   55987   6   &gt;2       29   2046   n/a   55956   6   &gt;2       31   1912   n/a   55952   6   &gt;2       32   2064   n/a   55957   6   &gt;2       34   1502/3852   n/a   55559   6   &gt;2       11   2814   n/a   55978   6   &gt;2       13   3478   n/a   55980   6   &gt;2       17    553   n/a   55561   6   &gt;2       19   3275   n/a   55984   6   &gt;2       21   2809   n/a   58261   6   &gt;2       23   1677   n/a   58348   6   &gt;2                  
 
     [0406] Each of the sequences was then used as a query to search the public databases in order to facilitate identification of extended sequences for these clones. Extended sequence information for the above sequences, obtained by searching public sequence databases, is set forth in SEQ ID NO:2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 33, and 35, respectively.  
     Example 3  
     Quantitative Real-time RT-PCR Analysis  
     [0407] Briefly, quantitation of PCR product relies on the few cycles where the amount of DNA amplifies logarithmically from barely above the background to the plateau. Using continuous fluorescence monitoring, the threshold cycle number where DNA amplifies logarithmically is easily determined in each PCR reaction. There are two fluorescence detecting systems. One is based upon a double-strand DNA specific binding dye SYBR Green I dye. The other uses TaqMan probe containing a Reporter dye at the 5′ end (FAM) and a Quencher dye at the 3′ end (TAMRA) (Perkin Elmer/Applied Biosystems Division, Foster City, Calif.). Target-specific PCR amplification results in cleavage and release of the Reporter dye from the Quencher-containing probe by the nuclease activity of AmpliTaq Gold™ (Perkin Elmer/Applied Biosystems Division, Foster City, Calif.). Thus, fluorescence signal generated from released reporter dye is proportional to the amount of PCR product. Both detection methods have been found to generate comparable results. To compare the relative level of gene expression in multiple tissue samples, a panel of cDNAs is constructed using RNA from tissues and/or cell lines, and Real-Time PCR is performed using gene specific primers to quantify the copy number in each cDNA sample. Each cDNA sample is generally performed in duplicate and each reaction repeated in duplicated plates. The final Real-time PCR result is typically reported as an average of copy number of a gene of interest normalized against internal actin number in each cDNA sample. Real-time PCR reactions may be performed on a GeneAmp 5700 Detector using SYBR Green I dye or an ABI PRISM 7700 Detector using the TaqMan probe (Perkin Elmer/Applied Biosystems Division, Foster City, Calif.).  
     [0408] Using this approach, Real Time PCRE profiles were generated for L1027, L1037, L1038, L1039, L1040 and L1041, and are provided in Table 3.  
                       TABLE 3                       SEQ ID   CLONE           NO:   NAME   REAL TIME PROFILE                                            9   L1027C   Real Time PCR shows over-expression in small cell               lung carcinoma as well as in bone marrow. Expression               is also observed for multiple normal tissue.       5   L1037C   Real Time PCR shows over-expression in small cell               lung carcinoma as well as in bone marrow and lymph               node. Expression is also observed for multiple normal               tissue.       7   L1038C   Real Time PCR shows over-expression in small cell               lung carcinoma as well as in brain, pituitary gland and               adrenal gland. Expression is also observed for multiple               normal tissue.       3   L1039C   Real Time PCR shows over-expression in small cell               lung carcinoma as well as in lymph node. Expression               is also observed for multiple normal tissue.       1   L1040C   Real Time PCR shows over-expression in small cell               lung carcinoma as well as in brain, pituitary gland and               adrenal gland. Expression is also observed for multiple               normal tissue.       15   L0141C   Real Time PCR shows over-expression in small cell               lung carcinoma as well as in adrenal gland, bone               marrow and thymus. Expression is also observed for               multiple normal tissue.                  
 
     Example 4  
     Cloning of Full-length cDNA Sequences and ORF for L1027C  
     [0409] cDNA sequences encoding the full-length sequence for L1027C were isolated by screening a small cell primary tumor full length cloning library with a radioactively labeled probe of the original isolate sequence (SEQ ID NO:9). In order to determine the transcript size of the gene, a multiple tissue Northern blot was probed with the radioactively labelled original isolate sequence, SEQ ID NO:9. The Northern blot included 1 μg of small cell primary tumor polyA+ RNA. Visual analysis of the exposed film revealed a single transcript of approximately 2.5 kb. Approximately 500,000 clones from the full-length cloning library were screened and four clones were obtained from this library. The inserts were sequenced and yielded DNA nucleotide molecules of about 2.32 and 2.37 kb. These sequences are provided in SEQ ID NO:93 and 94, respectively. Both of these sequences contain the same single OFR of 450 bp (SEQ ID NO:95), and encode a deduced amino acid sequence of 150 amino acid residues (SEQ ID NO:96). These sequences were searched against the Genbank nonredundant and GeneSeq DNA databases and showed no hits.  
     Example 5  
     Analysis of cDNA Expression Using Microarray Technology  
     [0410] An additional 5054 of the resulting clones obtained from the lung electronic subtraction of Example 1 were probed by microarray chip technology to further characterize the expression of these clones. The microarray analysis was carried out as provided in Example 2. The clones were arrayed on Lung Chip 7. CorixArray analysis was performed on the microarray results to compare expression in lung tumors and in normal tissues. Clones were selected based on two criteria: 2-fold overexpression in lung tumors when compared to non-lung tissue and a mean expression level of less than 0.2 in these same non-lung tissues. Of those analyzed, 2372 clones met the criteria.  
     [0411] Microarray analysis for five of these clones is presented in Table 4:  
                                   TABLE 4                                       MICRO-   MICRO-           SEQ ID           ARRAY   ARRAY RATIO       SEQ   NO:           ANALYSIS   (Lung       ID   from   Clone   Clone   (Lung   Tumor:Normal       NO:   60/207,485   Name:   ID #   Chip #)   Tissue)                                                        42   18618   L1053C   63575   7   13.5       43   14788   L1054C   63582   7   5.29       44    7744   L1055C   63598   7   15.25       45    4257   L1056C   64963   7   9.31       46   20087   L1058C   64988   7   5.66                  
 
     Example 6  
     Quantitative Real-time PCR Analysis  
     [0412] 170 of the 2372 clones of Example 4 were further analyzed by visual analysis based on high expression in tumors and little or no expression in normal tissues. Seven clones were selected for Real-time PCR analysis. The Real-time PCR was carried out as disclosed in Example 3. The Real-time PCR profiles of these seven clones are presented in Table 5. The sequences of these seven clones are provided in SEQ ID NO:42-48, respectively.  
                           TABLE 5                       SEQ ID   CLONE   CLONE           NO:   NAME   ID #   REAL TIME PROFILE                  42   L1053C   63575   Real Time PCR shows over-expression in                   small cell lung carcinoma as well as in                   pituitary. Expression is also observed for                   multiple normal tissues.       43   L1054C   63582   Real Time PCR shows over-expression in                   small cell lung carcinoma as well as in                   pituitary, brain and spinal cord. Expression is                   also observed for adrenal and pancreas.       44   L1055C   63598   Real Time PCR shows over-expression in                   small cell lung carcinoma as well as in                   pituitary and brain. Expression is also                   observed for multiple normal tissues.       45   L1056C   64963   Real Time PCR shows over-expression in                   one small cell lung carcinoma sample. No                   expression is otherwise observed.       46   L1058C   64988   Real Time PCR shows over-expression in                   small cell lung carcinoma. Low level                   expression is also observed for adrenal                   gland, pancreas, and bone marrow.       47   n/a   63485   Real Time PCR shows over-expression in                   metastatic tumor as well as low level                   expression in multiple normal tissues.       48   n/a   65010   Real Time PCR shows low expression in one                   lung sample. No expression is otherwise                   observed.                  
 
     [0413] Each of the sequences was then used as a query to search the public databases in order to facilitate identification of extended sequences for these clones. SEQ ID NO:42, 43 and 45 matched to known genes in Genbank, and these results are presented in Table 6. The full-length cDNA sequences of the known genes are disclosed in SEQ ID NO:49, 50 and 52, respectively. The deduced amino acid sequences encoded by SEQ ID NO:49 and 50 are also provided as SEQ ID NO:56 and 57, respectively. SEQ ID NO:44 and 46-48 were found to be novel with respect to known genes, but matched to public EST sequences. The sequences of SEQ ID NO:44 and 46-48 were aligned with the matching EST sequences in order to obtain extended sequence data. These extended sequences are provided in SEQ ID NO:51 and 53-55, respectively.  
                               TABLE 6                                   SEQ ID NO:   CLONE NAME   GENBANK DESCRIPTION                          42   L1053C   Insulinoma-associated 1           43   L1054C   KIAA0535           45   L1056C   Human DAZ mRNA 3′ UTR                      
 
     Example 7  
     Cloning of cDNA Encoding Full-length L 1058C  
     [0414] The cDNA sequence encoding full-length L1058C was isolated by screening a small cell primary tumor full length cloning library with a radioactively labeled probe of the original isolate sequence (SEQ ID NO:46). In order to determine the transcript size of the gene, a multiple tissue Northern blot was probed with the radioactively labelled original isolate sequence, SEQ ID NO:46. The Northern blot included 1 μg of small cell primary tumor, carcinoid metastasis and small cell (tumor) cell line polyA+ RNA. Visual analysis of the exposed film revealed a single transcript of approximately 2.5 kb. Approximately 500,000 clones from the full-length cloning library were screened and one clone was obtained from this library. The insert was sequenced and yields a 2165 bp DNA nucleotide molecule. The full-length sequence is provided in SEQ ID NO:58. The full-length sequence is predicted to have two ORFs. A first ORF (SEQ ID NO:59) is predicted to encode a polypeptide having 392 amino acid residues (SEQ ID NO:61), and the second ORF (SEQ ID NO:60) is predicted to encode a polypeptide of 363 amino acid residues (SEQ ID NO:62) but does not show the starting methionine. This 2165 bp DNA was searched against the Genbank nonredundant and GeneSeq DNA databases and showed no hits.  
     Example 8  
     Analysis of cDNA Expression Using Microarray Technology  
     [0415] An additional 3453 of the resulting clones obtained from the lung electronic subtraction of Example 1 were probed by microarray chip technology to further characterize the expression of these clones. The microarray analysis was carried out as provided in Example 2. The clones were arrayed on Lung Chip 8. CorixArray analysis was performed on the microarray results to compare expression in lung tumors and in normal tissues. Clones were selected based on two criteria: 2-fold overexpression in lung tumors when compared to non-lung tissue and a mean expression level of less than 0.2 in these same non-lung tissues. Of those analyzed, 557 clones met the criteria.  
     [0416] 300 of the 557 clones were visually analyzed for overexpression in tumor versus normal tissue. Twenty-eight clones showing overexpression in tumor versus normal tissue were then sequenced. These DNA sequences are provided in SEQ ID NO:63-92, respectively. The microarray analysis for these 28 clones is presented in Table 7.  
                               TABLE 7                                   MEDIAN   MEDIAN       SEQ ID NO:   CLONE ID #   RATIO   SIGNAL 1   SIGNAL 2                                                    63   72761   2.22   0.154   0.07       64   72762   2.33   0.105   0.045       65   72763   2.41   0.233   0.097       66   72764   2.72   0.199   0.073       67   72765   2.62   0.158   0.06       68   72766   2.84   0.149   0.053       69   72772   2.25   0.109   0.049       70   72775   2.36   0.103   0.044       71   72776   2.34   0.146   0.062       72   72779   2.25   0.22   0.098       73   72781   2.51   0.149   0.059       74   72784   2.35   0.212   0.09       75   72788   2.85   0.152   0.053       76   72789   2.69   0.196   0.073       77   72790   2.46   0.181   0.074       78   72791   2.39   0.143   0.06       79   72792   2.43   0.197   0.081       80   72794   3.04   0.258   0.085       81   72795   2.37   0.143   0.06       82   72797   2.96   0.233   0.079       83   72798   2.82   0.218   0.077       84   72804   2.33   0.14   0.06       85   72805   2.33   0.102   0.043       86   72806   2.32   0.121   0.052       87   72807   3.02   0.117   0.039       88   72808   2.74   0.109   0.04       89   72809   2.26   0.126   0.056       90   72811   2.92   0.151   0.052       91   72813   2.66   0.138   0.052           (L1080C)                  
 
     [0417] Each of the sequences was then used as a query to search the public sequence databases to identify novel and known genes. Results of this search are provided in Table 8.  
                           TABLE 8                       SEQ ID   GEN BANK               NO:   ACC #   GENESEQ   DESCRIPTION                  63   AC004590       Chromosome 17       64   Z78409   T62661   transcription factor E2F5       65   S45828   Z86797;   cDNA DKFZp564L2416;               A09328   nekl = serine/threonine-and                   tyrosine-specific protein kinase                   [mice, erythroleukemia cells]       66           Novel       67   AL136169       Chromosome Xq26.1-27.1       68   AC011742       Chromosome 2,           AK021426         Homo sapiens  cDNA FLJ11364                   fis. clone HEMBA 1000264.       69   NM 005414   Q03742   SKI-like (SKIL)       70   NM 002335   V85551   low density lipoprotein receptor-                   related protein 5       71   XM_004587         Homo sapiens  adaptor protein                   with pleckstrin homology and src                   homology 2 domains (APS),           AB000520       mRNA.                     Homo sapiens  mRNA for APS,                   complete cds.       72   AK024119       cDNA FLJ14057 fis, clone                   HEMBB 1000337.       73   U86338       Mus musculus zinc finger protein                   Png-1 (Png-1)       74           Novel       75           Novel       76   NM_002271   C03734     Homo sapiens  karyopherin                   (importin) beta 3 (KPNB3) mRNA       77   NM_001401   T48669;     Homo sapiens  endothelial               T44104   differentiation, lysophosphatidic                   acid G-protein-coupled receptor,                   2(EDG2), mRNA.       78   U40583       Human alpha/neuronal nicontinic                   acetylcholine receptor mRNA,                   complete cds.       79       Z15509   Novel       80   Z59860   V34162     H. sapiens  CpG island DNA                   genomic Msel fragment, clone                   178c7, reverse read                   cpg178c7.rtla.       81           Novel       82   Z59860   HNGIT2   DNA genomic Msel fragment,               2   clone 178c7       83   XM-004477   Q72451     Homo sapiens  glutamate-cysteine                   ligase, catalytic subunit (GCLC),                   mRNA.       84       Z16421   Novel       85           Novel       86   AC022013   V52850   Chromosome 3       87           Novel       88   AL354993   Z91766   Chromosome 20q13.2-13.                   Continas a peptidylprolyl                   isomerase A (cyclophilin A)                   pseudogene, the gene for                   OVC10-2, ESTs, STSs and                   GSSs, complete sequence       89   AC005021       Chromosome 7q21-q22, complete                   sequence.       90   AK023904       cDNA FLJ13842 fis, clone                   THYRO1000793.                  
 
     Example 9  
     Quantitative Real-time PCR Analysis  
     [0418] One of the clones of Example 7, clone L1080C, was further selected for Real-time PCR analysis. The Real-time PCR was carried out as disclosed in Example 3. The Real-time PCR shows over-expression in small cell lung carcinoma as well as in brain and pituitary. Expression was also observed in thyroid, adrenal and salivary glands.  
     Example 10  
     Identifying Full-length cDNA Sequence Encoding L1080C  
     [0419] The cDNA sequence encoding full-length L1080C was predicted by using a partial sequence as a query to search the public sequence databases to obtain extended sequence. The query resulted in the identification of a full-length cDNA sequence for L1080C (SEQ ID NO:91). The deduced amino acid sequence encoded by the full-length cDNA sequence is provided in SEQ ID NO:92.  
     Example 11  
     Peptide Priming of T-helper Lines  
     [0420] Generation of CD4 +  T helper lines and identification of peptide epitopes derived from tumor-specific antigens that are capable of being recognized by CD4 +  T cells in the context of HLA class II molecules, is carried out as follows:  
     [0421] Fifteen-mer peptides overlapping by 10 amino acids, derived from a tumor-specific antigen, are generated using standard procedures. Dendritic cells (DC) are derived from PBMC of a normal donor using GM-CSF and IL-4 by standard protocols. CD4 +  T cells are generated from the same donor as the DC using MACS beads (Miltenyi Biotec, Auburn, Calif.) and negative selection. DC are pulsed overnight with pools of the 15-mer peptides, with each peptide at a final concentration of 0.25 μg/ml. Pulsed DC are washed and plated at 1×10 4  cells/well of 96-well V-bottom plates and purified CD4 +  T cells are added at 1×10 5 /well. Cultures are supplemented with 60 ng/ml IL-6 and 10 ng/ml IL-12 and incubated at 37° C. Cultures are restimulated as above on a weekly basis using DC generated and pulsed as above as antigen presenting cells, supplemented with 5 ng/ml IL-7 and 10 U/ml IL-2. Following 4 in vitro stimulation cycles, resulting CD4+T cell lines (each line corresponding to one well) are tested for specific proliferation and cytokine production in response to the stimulating pools of peptide with an irrelevant pool of peptides used as a control.  
     Example 12  
     Generation of Tumor-specific CTL Lines Using in Vitro Whole-gene Priming  
     [0422] Using in vitro whole-gene priming with tumor antigen-vaccinia infected DC (see, for example, Yee et al,  The Journal of Immunology,  157(9):4079-86, 1996), human CTL lines are derived that specifically recognize autologous fibroblasts transduced with a specific tumor antigen, as determined by interferon-γ ELISPOT analysis. Specifically, dendritic cells (DC) are differentiated from monocyte cultures derived from PBMC of normal human donors by growing for five days in RPMI medium containing 10% human serum, 50 ng/ml human GM-CSF and 30 ng/ml human IL-4. Following culture, DC are infected overnight with tumor antigen-recombinant vaccinia virus at a multiplicity of infection (M.O.I) of five, and matured overnight by the addition of 3 μg/ml CD40 ligand. Virus is then inactivated by UV irradiation. CD8 +  T cells are isolated using a magnetic bead system, and priming cultures are initiated using standard culture techniques. Cultures are restimulated every 7-10 days using autologous primary fibroblasts retrovirally transduced with previously identified tumor antigens. Following four stimulation cycles, CD8 +  T cell lines are identified that specifically produce interferon-γ when stimulated with tumor antigen-transduced autologous fibroblasts. Using a panel of HLA-mismatched B-LCL lines transduced with a vector expressing a tumor antigen, and measuring interferon-γ production by the CTL lines in an ELISPOT assay, the HLA restriction of the CTL lines is determined.  
     Example 13  
     Generation and Characterization of Anti-tumor Antigen Monoclonal Antibodies  
     [0423] Mouse monoclonal antibodies are raised against E. coli derived tumor antigen proteins as follows: Mice are immunized with Complete Freund&#39;s Adjuvant (CFA) containing 50 μg recombinant tumor protein, followed by a subsequent intraperitoneal boost with Incomplete Freund&#39;s Adjuvant (IFA) containing 10 μg recombinant protein. Three days prior to removal of the spleens, the mice are immunized intravenously with approximately 50 μg of soluble recombinant protein. The spleen of a mouse with a positive titer to the tumor antigen is removed, and a single-cell suspension made and used for fusion to SP2/O myeloma cells to generate B cell hybridomas. The supernatants from the hybrid clones are tested by ELISA for specificity to recombinant tumor protein, and epitope mapped using peptides that span the entire tumor protein sequence. The mAbs are also tested by flow cytometry for their ability to detect tumor protein on the surface of cells stably transfected with the cDNA encoding the tumor protein.  
     Example 14  
     Synthesis of Polypeptides  
     [0424] Polypeptides are synthesized on a Perkin Elmer/Applied Biosystems Division 430A peptide synthesizer using FMOC chemistry with HPTU (O-Benzotriazole-N,N,N′,N′-tetramethyluronium hexafluorophosphate) activation. A Gly-Cys-Gly sequence is attached to the amino terminus of the peptide to provide a method of conjugation, binding to an immobilized surface, or labeling of the peptide. Cleavage of the peptides from the solid support is carried out using the following cleavage mixture: trifluoroacetic acid:ethanedithiol:thioanisole:water:phenol (40:1:2:2:3). After cleaving for 2 hours, the peptides are precipitated in cold methyl-t-butyl-ether. The peptide pellets are then dissolved in water containing 0.1% trifluoroacetic acid (TFA) and lyophilized prior to purification by C18 reverse phase HPLC. A gradient of 0%-60% acetonitrile (containing 0.1% TFA) in water (containing 0.1% TFA) is used to elute the peptides. Following lyophilization of the pure fractions, the peptides are characterized using electrospray or other types of mass spectrometry and by amino acid analysis.  
     [0425] From the foregoing it will be appreciated that, although specific embodiments of the invention have been described herein for purposes of illustration, various modifications may be made without deviating from the spirit and scope of the invention. Accordingly, the invention is not limited except as by the appended claims.  
    
     
       
         1 
         
           
             96  
           
           
             1  
             644  
             DNA  
             Homo sapiens  
             
               misc_feature  
               (1)...(644)  
               n = A,T,C or G  
             
           
            1 

ttactcctct agagggaaag catgacaccg aacactaagc acacagcttt ttgttgtttt     60 

ggttttttct cccgcaaatc ttaaagtgat tcccatgacc ttggccaagg acacttctta    120 

aagattaatg actggcactg acattgcccc aggcgggcca ctcctcacac tggctctcag    180 

ttcccagcca tgcctggggc tcagtcactt ctattccacc ctctgagact ccattggtgt    240 

cacacaaggt gtcttcttgg ctttgatttt gagaatcccc tattttcact tccagatctg    300 

tcagctgcca tggaggaata atagaaaacc agaaatgcgt gtagagggag atttctaaaa    360 

cttcccttgt gtcgccatag ttgtagtttt gggttctggc aggtggaaca ccctgaaacc    420 

tggaatcatt ctatgagaat acagttcaga ctttgcagac tccagcccat actaactgtc    480 

atgaagcttg acttcttgtc ataatgcagc catcttggag gaaattggca tttctgctta    540 

gatggntggc agggtcgcgc tcagctttgc tttctacact aaattacata gcattaattc    600 

aagnattgtt ttccaatttc ccatccctga tttccagctt tctt                     644 

 
           
             2  
             1115  
             DNA  
             Homo sapiens  
           
            2 

gtaggaagtt acagtaaatg gtagttcatt cttacttaca cacatagcta atcttttttt     60 

tttcacttgg aattatgttg aatgtttcat tttgacaaaa aagtagacta gaaggtatgt    120 

yctttaagtt gtcttgcatc cattatataa gaaagaaaca ggtgagagga agagcagaaa    180 

gctgagactg gctgatgttc agagcactta ctcctctaga gggaaagcat gacaccgaac    240 

actaagcaca cagctttttg ttgttttggt tttttctccc gcaaatctta aagtgattcc    300 

catgaccttg gccaaggaca cttcttaaag attaatgact ggcactgaca ttgccccagg    360 

cgggccactc ctcacactgg ctctcagttc ccagccatgc ctggggctca gtcacttcta    420 

ttccaccctc tgagactcca ttggtgtcac acaaggtgtc ttcttggctt tgattttgag    480 

aatcccctat tttcacttcc agatctgtca gctgccatgg aggaataata gaaaaccaga    540 

aatgcgtgta gagggagatt tctaaaactt cccttgtgtc gcccatagtt gtagttttgg    600 

gttctggcag gtggaacacc ctgaaacctg gaatcattct atgagaatac agttcagact    660 

ttgcagactc cagcccatac taactgtcat gaagcttgac ttcttgtcat aatgcagcca    720 

tcttggagga aattggccat ttctgcttag atggttggca gggtcgcgct cagctttgct    780 

ttctacacta attacatagc attattcaag tattgttttc catttcccat ccctgatttc    840 

cagcttctta aagctgactg ttcttgcagg ggccacttgc ttctcctaga gtacaaaagt    900 

aagggccttc cttactaact gcagggtctc tctattacac ctcaacatac acactttgct    960 

gctactgttt gtactgtcta cagtagaatt tccttatctt gctcctggta gtgcattaca   1020 

ggcaagcatg aaatgtaaag tatttattta aataaaaaga aaacctctaa attggtaatt   1080 

gaawwammwm mmwrwarmww tatagtttgt gacat                              1115 

 
           
             3  
             540  
             DNA  
             Homo sapiens  
           
            3 

gggccagaat tcggccgagg cctgcaaacg agaaggctgt ggatttgatt attgtacgaa     60 

gtgtctctgt aattatcata ctactaaaga ctgttcagat ggcaagctcc tcaaagccag    120 

ttgtaaaata ggtcccctgc ctggtacaaa gaaaagcaaa aagaatttac gaagattgtg    180 

atctcttatt aaatcaattg ttactgatca tgaatgttag ttagaaaatg ttaggtttta    240 

acttaaaaaa aattgtattg tgattttcaa ttttatgttg aaatcggtgt agtatcctga    300 

ggtttttttc cccccagaag ataaagagga tagacaacct cttaaaatat ttttacaatt    360 

taatgagaaa aagtttaaaa ttctcaatac aaatcaaaca atttaaatat tttaagaaaa    420 

aaggaaaagt agatagtgat actgagggta aaaaaaaatt gattcaattt tatggtaaag    480 

gaaacccatg caattttacc tagacagtct taaatatgtc tggttttcca tctgttagca    540 

 
           
             4  
             2076  
             DNA  
             Homo sapiens  
           
            4 

aggttgctca gctgcccccg gagcggttcc tccacctgag gcagacacca cctcggttgg     60 

catgagccgg cgcccctgca gctgcgccct acggccaccc cgctgctcct gcagcgccag    120 

ccccagcgca gtgacagccg ccgggcgccc tcgaccctcg gatagttgta aagaagaaag    180 

ttctaccctt tctgtcaaaa tgaagtgtga ttttaattgt aaccatgttc attccggact    240 

taaactggta aaacctgatg acattggaag actagtttcc tacacccctg catatctgga    300 

aggttcctgt aaagactgca ttaaagacta tgaaaggctg tcatgtattg ggtcaccgat    360 

tgtgagccct aggattgtac aacttgaaac tgaaagcaag cgcttgcata acaaggaaaa    420 

tcaacatgtg caacagacac ttaatagtac aaatgaaata gaagcactag agaccagtag    480 

actttatgaa gacagtggct attcctcatt ttctctacaa agtggcctca gtgaacatga    540 

agaaggtagc ctcctggagg agaatttcgg tgacagtcta caatcctgcc tgctacaaat    600 

acaaagccca gaccaatatc ccaacaaaaa cttgctgcca gttcttcatt ttgaaaaagt    660 

ggtttgttca acattaaaaa agaatgcaaa acgaaatcct aaagtagatc gggagatgct    720 

gaaggaaatt atagccagag gaaattttag actgcagaat ataattggca gaaaaatggg    780 

cctagaatgt gtagatattc tcagcgaact ctttcgaagg ggactcagac atgtcttagc    840 

aactatttta gcacaactca gtgacatgga cttaatcaat gtgtctaaag tgagcacaac    900 

ttggaagaag atcctagaag atgataaggg ggcattccag ttgtacagta aagcaataca    960 

aagagttacc gaaaacaaca ataaattttc acctcatgct tcaaccagag aatatgttat   1020 

gttcagaacc ccactggctt ctgttcagaa atcagcagcc cagacttctc tcaaaaaaga   1080 

tgctcaaacc aagttatcca atcaaggtga tcagaaaggt tctacttata gtcgacacaa   1140 

tgaattctct gaggttgcca agacattgaa aaagaacgaa agcctcaaag cctgtattcg   1200 

ctgtaattca cctgcaaaat atgattgcta tttacaacgg gcaacctgca aacgagaagg   1260 

ctgtggattt gattattgta cgaagtgtct ctgtaattat catactacta aagactgttc   1320 

agatggcaag ctcctcaaag ccagttgtaa aataggtccc ctgcctggta caaagaaaag   1380 

caaaaagaat ttacgaagat tgtgatctct tattaaatca attgttactg atcatgaatg   1440 

ttagttagaa aatgttaggt tttaacttaa aaaaaattgt attgtgattt tcaattttat   1500 

gttgaaatcg gtgtagtatc ctgaggtttt tttcccccca gaagataaag aggatagaca   1560 

acctcttaaa atatttttac aatttaatga gaaaaagttt aaaattctca atacaaatca   1620 

aacaatttaa atattttaag aaaaaaggaa aagtagatag tgatactgag ggtaaaaaaa   1680 

aaattgattc aattttatgg taaaggaaac ccatgcaatt ttacctagac agtcttaaat   1740 

atgtctggtt ttccatctgt tagcatttca gacattttat gttcctctta ctcaattgat   1800 

accaacagaa atatcaactt ctggagtcta ttaaatgtgt tgtcaccttt ctaaagcttt   1860 

ttttcattgt gtgtatttcc caagaaagta tcctttgtaa aaacttgctt gttttcctta   1920 

tttctgaaat ctgttttaat atttttgtat acatgtaaat atttctgtat tttttatatg   1980 

tcaaagaata tgtctcttgt atgtacatat aaaaataaat tttgctcaat aaaattgtaa   2040 

gcttaaaaaa aaaaaaaaaa aactcgagac tagtgc                             2076 

 
           
             5  
             634  
             DNA  
             Homo sapiens  
           
            5 

gggcagaatt cggacgagga cttttcctca gtgttgacct tagggtgcag ctggatgttt     60 

ttaccctcag cggctttcgg actgtacaga tcctggaagg acaaaagatc ctggctaact    120 

gttcttctcc ctaccaggta gacctgtttg gtatagcaga tttagcacat ttactattgt    180 

tcaaggaaca cctacaggtc ttctgggatg ggtccttctg gaaacttagc caaaatattt    240 

ctgagctaaa agatggtgaa ttgtggaata aattctttgt gcggattctg aatgccaatg    300 

atgaggccac agtgtctgtt cttggggagc ttgcagcaga aatgaatggg gtttttgaca    360 

ctacattcca aagtcacctg aacaaagcct tatggaaggt agggaagtta actagtcctg    420 

gggctttgct ctttcagtga gctaggcaat caagtctcac agattgctgc ctcagagcaa    480 

tggttgtatt gtggaacact gaaactgtat gtgctgtaat ttaatttagg acacatttag    540 

atgcactacc attgctgttc tactttttgg tacaggtata ttttgacgtc actgatattt    600 

tttatacagt gatatactta ctcatggcct tgct                                634 

 
           
             6  
             3725  
             DNA  
             Homo sapiens  
           
            6 

accgttaaat ttgaaacttg gcgggtaggg gtgtgggctt gaggtggccg gtttgttagg     60 

gagtcgtgtg cgtgccttgg tcgcttctgt agctccgagg gcaggttgcg gaagaaagcc    120 

caggcggtct gtggcccaga ggaaaggcct gcagcaggac gaggacctga gccaggaatg    180 

caggatggcg gcggtgaaga aggaaggggg tgctctgagt gaagccatgt ccctggaggg    240 

agatgaatgg gaactgagta aagaaaatgt acaaccttta aggcaagggc ggatcatgtc    300 

cacgcttcag ggagcactgg cacaagaatc tgcctgtaac aatactcttc agcagcagaa    360 

acgggcattt gaatatgaaa ttcgatttta cactggaaat gaccctctgg atgtttggga    420 

taggtatatc agctggacag agcagaacta tcctcaaggt gggaaggaga gtaatatgtc    480 

aacgttatta gaaagagctg tagaagcact acaaggagaa aaacgatatt atagtgatcc    540 

tcgatttctc aatctctggc ttaaattagg gcgtttatgc aatgagcctt tggatatgta    600 

cagttacttg cacaaccaag ggattggtgt ttcacttgct cagttctata tctcatgggc    660 

agaagaatat gaagctagag aaaactttag gaaagcagat gcgatatttc aggaagggat    720 

tcaacagaag gctgaaccac tagaaagact acagtcccag caccgacaat tccaagctcg    780 

agtgtctcgg caaactctgt tggcacttga gaaagaagaa gaggaggaag tttttgagtc    840 

ttctgtacca caacgaagca cactagctga actaaagagc aaagggaaaa agacagcaag    900 

agctccaatc atccgtgtag gaggtgctct caaggctcca agccagaaca gaggactcca    960 

aaatccattt cctcaacaga tgcaaaataa tagtagaatt actgtttttg atgaaaatgc   1020 

tgatgaggct tctacagcag agttgtctaa gcctacagtc cagccatgga tagcaccccc   1080 

catgcccagg gccaaagaga atgagctgca agcaggccct tggaacacag gcaggtcctt   1140 

ggaacacagg cctcgtggca atacagcttc actgatagct gtacccgctg tgcttcccag   1200 

tttcactcca tatgtggaag agactgcaca acagccagtt atgacaccat gtaaaattga   1260 

acctagtata aaccacatcc taagcaccag aaagcctgga aaggaagaag gagatcctct   1320 

acaaagggtt cagagccatc agcaagcatc tgaggagaag aaagagaaga tgatgtattg   1380 

taaggagaag atttatgcag gagtagggga attctccttt gaagaaattc gggctgaagt   1440 

tttccggaag aaattaaaag agcaaaggga agccgagcta ttgaccagtg cagagaagag   1500 

agcagaaatg cagaaacaga ttgaagagat ggagaagaag ctaaaagaaa tccaaactac   1560 

tcagcaagaa agaacaggtg atcagcaaga agagacgatg cctacaaagg agacaactaa   1620 

actgcaaatt gcttccgagt ctcagaaaat accaggaatg actctatcca gttctgtttg   1680 

tcaagtaaac tgttgtgcca gagaaacttc acttgcggag aacatttggc aggaacaacc   1740 

tcattctaaa ggtcccagtg tacctttctc catttttgat gagtttcttc tttcagaaaa   1800 

gaagaataaa agtcctcctg cagatccccc acgagtttta gctcaacgaa gaccccttgc   1860 

agttctcaaa acctcagaaa gcatcacctc aaatgaagat gtgtctccag atgtttgtga   1920 

tgaatttaca ggaattgaac ccttgagcga ggatgccatt atcacaggct tcagaaatgt   1980 

aacaatttgt cctaacccag aagacacttg tgactttgcc agagcagctc gttttgtatc   2040 

cactcctttt catgagataa tgtccttgaa ggatctccct tctgatcctg agagactgtt   2100 

accggaagaa gatctagatg taaagacctc tgaggaccag cagacagctt gtggcactat   2160 

ctacagtcag actctcagca tcaagaagct gagcccaatt attgaagaca gtcgtgaagc   2220 

cacacactcc tctggcttct ctggttcttc tgcctcggtt gcaagcacct cctccatcaa   2280 

atgtcttcaa attcctgaga aactagaact tactaatgag acttcagaaa accctactca   2340 

gtcaccatgg tgttcacagt atcgcagaca gctactgaag tccctaccag agttaagtgc   2400 

ctctgcagag ttgtgtatag aagacagacc aatgcctaag ttggaaattg agaaggaaat   2460 

tgaattaggt aatgaggatt actgcattaa acgagaatac ctaatatgtg aagattacaa   2520 

gttattttgg gtggcgccaa gaaactttgc agaattaaca gtaataaagg tatcttctca   2580 

acctgtccca tgggactttt atatcaacct caagttaaag gaacgtttaa atgaagattt   2640 

tgatcatttt tgcagctgtt atcaatatca agatggctgt attgtttggc accaatatat   2700 

aaactgcttc acccttcagg atcttctcca acacagtgaa tatattaccc atgaaataac   2760 

agtgttgatt atttataacc ttttgacaat agtggagatg ctacacaaag cagaaatagt   2820 

ccatggtgac ttgagtccaa ggtgtctgat tctcagaaac agaatccacg atccctatga   2880 

ttgtaacaag aacaatcaag ctttgaagat agtggacttt tcctacagtg ttgaccttag   2940 

ggtgcagctg gatgttttta ccctcagcgg ctttcggact gtacagatcc tggaaggaca   3000 

aaagatcctg gctaactgtt cttctcccta ccaggtagac ctgtttggta tagcagattt   3060 

agcacattta ctattgttca aggaacacct acaggtcttc tgggatgggt ccttctggaa   3120 

acttagccaa aatatttctg agctaaaaga tggtgaattg tggaataaat tctttgtgcg   3180 

gattctgaat gccaatgatg aggccacagt gtctgttctt ggggagcttg cagcagaaat   3240 

gaatggggtt tttgacacta cattccaaag tcacctgaac aaagccttat ggaaggtagg   3300 

gaagttaact agtcctgggg ctttgctctt tcagtgagct aggcaatcaa gtctcacaga   3360 

ttgctgcctc agagcaatgg ttgtattgtg gaacactgaa actgtatgtg ctgtaattta   3420 

atttaggaca catttagatg cactaccatt gctgttctac tttttggtac aggtatattt   3480 

tgacgtcact gatatttttt atacagtgat atacttactc atggccttgt ctaacttttg   3540 

tgaagaacta ttttattcta aacagactca ttacaaatgg ttaccttgtt atttaaccca   3600 

tttgtctcta cttttccctg tacttttccc atttgtaatt tgtaaaatgt tctcttatga   3660 

tcaccatgta ttttgtaaat aataaaatag tatctgttaa aaaaaaaaaa aaaaaaaaaa   3720 

aaaaa                                                               3725 

 
           
             7  
             567  
             DNA  
             Homo sapiens  
             
               misc_feature  
               (1)...(567)  
               n = A,T,C or G  
             
           
            7 

ggccaagaat tcggcacgag gacaacatac taaagaggcg aggcaatgac tgttggccag     60 

ttctcaccgg ggaaaaaccc actgttagga tggcatgaac atttccttag atcgtggnca    120 

gctccgagga atgtggcgtn caggctcttt gagagccatg ggctgcaccc ggccgtaggc    180 

tagtgtaact cgcatcccat tgcagtgccg tttcttgact gtgttgctgt ctcttagatt    240 

aaccgtgctg aggctccaca tagctcctgg acctgtgtct agtacatact gaagcgatgg    300 

tcagagtgtg tagagtgaag ttgctgtgcc cacattgttt gaactcgcgt accccgtaga    360 

tacattgtgc aacgttcttc tgttattccc ttgaggtggt aacttcgtat gttcagttta    420 

tgcgatgatt gttgtaaatg caatgccgta gtttggatta ataagtggat ggtttttgtt    480 

tctaaaaaga aaaaaaaaat cagtgttcac ccttatagag acatagtcaa gttcatgttg    540 

ataataatca aaggaattac tctcttc                                        567 

 
           
             8  
             1365  
             DNA  
             Homo sapiens  
           
            8 

acttcatgaa cacggacaat ttcacctccc accgtctccc ccacccctgg tcgggcacgg     60 

ggcaggtggt ctacaacggt tctatctact ttaacaagtt ccagagccac atcatcatca    120 

ggtttgacct gaagacagag accatcctca agacccgcag cctggactat gccggttaca    180 

acaacatgta ccactacgcc tggggtggcc actcggacat cgacctcatg gtggacgaga    240 

gcgggctgtg ggccgtgtac gccaccaacc agaacgctgg caacatcgtg gtcagtaggc    300 

tggaccccgt gtccctgcag accctgcaga cctggaacac gagctacccc aagcgcagcg    360 

ccggggaggc cttcatcatc tgcggcacgc tgtacgtcac caacggctac tcagggggta    420 

ccaaggtcca ctatgcatac cagaccaatg cctccaccta tgaatacatc gacatcccat    480 

tccagaacaa atactcccac atctccatgc tggactacaa ccccaaggac cgggccctgt    540 

atgcctggaa caacggccac cagatcctct acaacgtgac cctcttccac gtcatccgct    600 

ccgacgagtt gtagctccct cctcctggaa gccaagggcc cacgtcctca ccacaaaggg    660 

actcctgtga aactgctgcc aaaaagatac caataacact aacaataccg atcttgaaaa    720 

atcatcagca gtgcggattc tgacatcgag ggatggcatt acctccgtgt ttctcccttt    780 

cgagccggcg ggccacagac gtcggaagaa actcccgtat ttgcagctgg aactgcagcc    840 

cacggcgccc cggttttcct ccccgccctg tccctctctg gtcaaacaac atactaaaga    900 

ggcgaggcaa tgactgttgg ccagttctca ccggggaaaa acccactgtt aggatggcat    960 

gaacatttcc ttagatcgtg gtcagctccg aggaatgtgg cgtccaggct ctttgagagc   1020 

catgggctgc acccggccgt aggctagtgt aactcgcatc ccattgcagt gccgtttctt   1080 

gactgtgttg ctgtctctta gattaaccgt gctgaggctc cacatagctc ctggacctgt   1140 

gtctagtaca tactgaagcg atggtcagag tgtgtagagt gaagttgctg tgcccacatt   1200 

gtttgaactc gcgtaccccg tagatacatt gtgcaacgtt cttctgttat tcccttgagg   1260 

tggtaacttc gtatgttcag tttatgcgat gattgttgta aatgcaatgc cgtagtttgg   1320 

attaataagt ggatggtttt tgtttctaaa aaaaaaaaaa aaaaa                   1365 

 
           
             9  
             1196  
             DNA  
             Homo sapiens  
           
            9 

ctcagctcta ggggaatgaa ggctgttttg ctggctgata ctgaaataga ccttttctct     60 

acagacatcc ctcctaccaa cgcagtggac ttcactggaa gatgctattt caccaaaatc    120 

tgcaaatgta aactgaagga catcgcatgt ttaaaatgtg ggaacattgt agtttatcat    180 

gtgattgttc catgtagttc ctgtcttctt tcctgcaaca acagacactt ctggatgttt    240 

cacagccagg cagtttatga tattaacaga ctagactcca caggtgtaaa cgtcctactt    300 

cggggcaact tgccagagat agaagagagt acagatgaag atgtgttaaa tatctcagca    360 

gaggagtgta ttagataaat ggaattatga tatatatgat atacaaactt ttttctattt    420 

aaaaatatat taatggatca actttaaaat tgttagttgc cagtgatctt ttttggaaaa    480 

caaaaatggg gcatttgttg atttatttat tttctgtctc taattagtta cctcagtttg    540 

attgaagcca gtggagttgt gcttttcctc tacttctact tcctctcccc cacctttttc    600 

tgcccagtgt aggtgtattc ttaaattcag acgggaagat tctttcacat atcactcagt    660 

tacctcccaa tctgggggag tttttcttac aacttgatac cagataccat taattttaca    720 

ttcctgaata aaggcctagt acccacgcat atttcaacca tgcatatatc aagttcaacy    780 

gagttttaat aggggattaa aaaaacaagc tgttaggttt ccatgggcac tggttctcat    840 

aggttctatt ggtgataact gctttaacat ggagcaagag tttgtgaatc aggaaataga    900 

ataaattaaa atttaaaata tatagaggaa tcctcttgat tgctcagcat gatgttagat    960 

aaatgagttt gtcagaaaat atcagtatac gctgtttacc aatgttattt atttacattc   1020 

ttctaaagcc attatggata ttgtattatg agagctaaac ctaaataagt tatcctgttc   1080 

cctaggacct tctctgtaaa tagtgaattt tagacgagta gtctgtccta aatcttaaat   1140 

agaaaaaaaa actaaagcga tttgcttaag ccattgtaca ttataaagag ctgttt       1196 

 
           
             10  
             1424  
             DNA  
             Homo sapiens  
           
            10 

ctcagctcta ggggaatgaa ggctgttttg ctggctgata ctgaaataga ccttttctct     60 

acagacatcc ctcctaccaa cgcagtggac ttcactggaa gatgctattt caccaaaatc    120 

tgcaaatgta aactgaagga catcgcatgt ttaaaatgtg ggaacattgt agkttatcat    180 

gtgattgttc catgtagttc ctgtcttctt tcctgcaaca acagacactt ctggatgttt    240 

cacagccagg cagtttatga tattaacaga ctagactcca caggtgtaaa cgtcctactt    300 

cggggcaact tgccagagat agaagagagt acagatgaag atgtgttaaa tatctcagca    360 

gaggagtgta ttagataaat ggaattatga tatatatgat atacaaactt ttttctattt    420 

aaaaatatat taatggatca actttaaaat tgttagttgc cagtgatctt tttkggaaaa    480 

caaaaatggg gcatttgttg atttatttat tttctgtctc taattagtta cctcagtttg    540 

attgaagcca gtggagttgt gcttttcctc tacttctact tcctctcccc cacctttttc    600 

tgcccagtgt aggtgtattc ttaaattcag acgggaagat tctttcacat atcactcagt    660 

tacctcccaa tctgggggag tttttcttac aacttgatac cagataccat taattttaca    720 

ttcctgaata aaggcctagt acccacgcat atttcaacca tgcatatatc aagttcaacy    780 

gagttttaat aggggattaa aaaaacaagc tgttaggttt ccatgggcac tggttctcat    840 

aggttctatt ggtgataact gctttaacat ggagcaagag tttgtgaatc aggaaataga    900 

ataaattaaa atttaaaata tatagaggaa tcctcttgat tgctcagcat gatgttagat    960 

aaatgagttt gtcagaaaat atcagtatac gctgtttacc aatgttattt atttacattc   1020 

ttctaaagcc attatggata ttgtattatg agagctaaac ctaaataagt tatcctgttc   1080 

cctaggacct tctctgtaaa tagtgaattt tagacgagta gtctgtccta aatcttaaat   1140 

agaaaaaaaa actaaagcga tttgcttaag ccattgtaca ttataaagag ctgttttgtt   1200 

ttgctttgct ttgctttgtt ttgttttttt taaagctgca ttcagagcca caaaggaata   1260 

ggaaagtagg gtagtgttgg attctggttt tatgtaactc taaaataaat gtatctcttt   1320 

aatatctcag ttgtagggat tttgtcaata ccaaagcaga ctgagttgtg gttttgtaaa   1380 

taaagttttt tctaaaaatg aaaaaaaaag aaaaaaaaaa aaaa                    1424 

 
           
             11  
             460  
             DNA  
             Homo sapiens  
             
               misc_feature  
               (1)...(460)  
               n = A,T,C or G  
             
           
            11 

agacagngac gtatggaaaa gntcttaaca gatnatttaa atgacctcca gggtcgcaat     60 

gatnatgacg ccagtggcac tngggacttc tatggggaca ntttgtttgt gaaccagatg    120 

atgaaagtgg caaggccaaa caggatncat ncgcctagag nagaanacna agatgatgat    180 

gacgatgcct atagcngatg tgtttgaatt ngaattttca gagacccccc tcttaccgtg    240 

ttataacatc caagtatctg tggctcaggg gccacgaaac tggctactgc tttcggatgt    300 

ccttaagaaa ttganaatgt cctcccgcat atttcgctgc anttttccaa acgnggaaat    360 

tgtcaccatt gcagaggcag aattttatcg gtaggtttct gcnagtctct tgntctcttg    420 

ctccaaagac ctggcaagcc ttcaaccctt gaaaggnaan                          460 

 
           
             12  
             2206  
             DNA  
             Homo sapiens  
           
            12 

cagaagacag atgtgctgtg tgcagacgaa gaagaggatt gccaggctgc ctccctgctg     60 

cagaaataca ccgacaacag cgagaagcca tccgggaaga gactgtgcaa aaccaaacac    120 

ttgatccctc aggagtccag gcggggattg ccactgacag gggaatacta cgtggagaat    180 

gccgatggca aggtgactgt ccggagattc agaaagcggc cggagcccag ttcggactat    240 

gatctgtcac cagccaagca ggagccaaag cccttcgacc gcttgcagca actgctacca    300 

gcctcccagt ccacacagct gccatgctca agttcccctc aggagaccac ccagtctcgc    360 

cctatgccgc cggaagcacg gagacttatt gtcagtaaga acgctggcga gacccttctg    420 

cagcgggcag ccaggcttgg ctatgaggaa gtggtcctgt actgcttaga gaacaagatt    480 

tgtgatgtaa atcatcggga caacgcaggt tactgcgccc tgcatgaagc ttgtgctagg    540 

ggctggctca acattgtgcg acacctcctt gaatatggcg ctgatgtcaa ctgtagtgcc    600 

caggatggaa ccaggcctct gcacgatgct gttgagaacg atcacttgga aattgtccga    660 

ctacttctct cttatggtgc tgaccccacc ttggctacgt actcaggtag aaccatcatg    720 

aaaatgaccc acagtgaact tatggaaagg ttcttaacag attatttaaa tgacctccag    780 

ggtcgcaatg atgatgacgc cagtggcact tgggacttct atggcagctc tgtttgtgaa    840 

ccagatgatg aaagtggcta tgatgtttta gccaaccccc caggaccaga agaccaggat    900 

gatgatgacg atgcctatag cgatgtgttt gaatttgaat tttcagagac ccccctctta    960 

ccgtgttata acatccaagt atctgtggct caggggtgag catggctgtc atgtgattga   1020 

aaactagctg agctgctctt gaggccacga aactggctac tgctttcgga tgtccttaag   1080 

aaattgaaaa tgtcctcccg catatttcgc tgcaattttc caaacgtgga aattgtcacc   1140 

attgcagagg cagaatttta tcggcaggtt tctgcaagtc tcttgttctc ttgctccaaa   1200 

gacctggaag ccttcaaccc tgaaagtaag gagctgttag atctggtgga attcacgaac   1260 

gaaattcaga ctctgctggg ctcctctgta gagtggctcc accccagtga tctggcctca   1320 

gacaactact ggtgagcaag ctggacccac catgtacagt gtgttatagt gttaatcctt   1380 

gtgcatatgt gtcataatac aactatttct gtaaagaaag gacactatta catatgaaaa   1440 

tatctcttct ttatataaga gaaattactc cagtcagaag gacttagaaa catgtttttt   1500 

tccttttaaa cttttaagtc agtttttatg aagttgttat aatgtttctt tacttttcaa   1560 

tgcacacatg ctttgggata cgtttgtttt tacttggaac atttgtttct tttctttttt   1620 

aaggagaaaa aaaaaatgag taaaaggagc tccacacttt gacttaattt catacaaagc   1680 

tctgatgaca ggccatgact gtagagtggt cagaactgtg tggttggttt gagggagcga   1740 

attcggggaa ggcacttggt gatataactt tgttttgttt acagagtacc tgctcgggcc   1800 

aggtaaatgc tattggatgt aatccagtag tgtgtaatat aaattcaaac catatccaca   1860 

cacaacaact aattgtatga aacttttata tcctaattta aaagctgtga aattagtttt   1920 

cacgcatcaa accggattgt ttatatgttt aaacatttta tgctcttatt taaagaagac   1980 

tttgagctat ttttttctgt accctgtaaa atattgaaaa ctaacataat atgttgaggt   2040 

tgcttggaaa tgtacataaa actaaaattt tctgaatcgt gtgtttatgt ttgaaatctg   2100 

tgttttaact ttgtaagtaa attctctgcc tttgtattta tattttacaa aattttctta   2160 

aaaggcataa aactgttgag gaaaggagaa aaaaaaaaaa aaaaaa                  2206 

 
           
             13  
             680  
             DNA  
             Homo sapiens  
             
               misc_feature  
               (1)...(680)  
               n = A,T,C or G  
             
           
            13 

ataagatccc agctttgcgg gaactcatgc actatctcag ggaggtgatg caggattacc     60 

gagatgagct caaggacttc tttgcagttg acaaacagct ggcatcagag cttgagtatg    120 

acatgaagaa gtaccaggaa cagctggtcc aggagcagga gctagcaaaa catgcagatg    180 

tggccgggac ggctggaggt gctgaggtgg cacctgtggc acaggttgcc ctgtgtttag    240 

aaacagtgcc agttcctgct ggccaagaaa accctgccat gtcacctgcc gtgagccagc    300 

cctgcacacc cagggcaagt gctggccatg tagcagtatc atctcctaca cctgaaacag    360 

ggccattgca gaggttgctg cccaaagcca ggcccatgtc cctgagcacc attgcaatcc    420 

tgaattctgt caagaaagcc gtggagtcaa agagcaggca tcggagtcgg agcttaggag    480 

tgctgccttt cactttaaat tctggaagcc cagaaaaaac gtgcagtcag gtgtcttcat    540 

acagtttgga gcaagagtcg aatggcgaga ttgagcacgt gaccaagcgg gccatcagca    600 

cccccgagaa gagcatcagt gatgtcacgt tttggagcan gggtcaagtt acatcgggac    660 

accacgggac ttccgtcgtc                                                680 

 
           
             14  
             5023  
             DNA  
             Homo sapiens  
           
            14 

ggcggcggcg agccggtgcc ctgggatcat ggtggcgttg cggggccttg gtagcggcct     60 

gcagccctgg tgtccgctgg atcttagact cgaatgggtt gacacagtgt gggaactgga    120 

tttcacagag actgagcctt tggatcccag catagaagca gagatcatag agactggatt    180 

ggctgcattc acaaaactct atgaaagcct tttacccttt gctactggag aacatggatc    240 

tatggagagt atctggacct tcttcattga gaacaatgtt tcccatagta cactggtggc    300 

attgttctat cattttgttc aaatagttca taagaagaat gtcagtgtac agtatcgaga    360 

atatggcctt catgccgctg ggctttactt tttgctacta gaagtaccag gcagtgtagc    420 

caatcaagta ttccacccag tgatgtttga caaatgcatt cagactctaa agaagagctg    480 

gccccaggaa tctaacttga atcggaaaag aaagaaagaa cagcctaaga gctctcaggc    540 

taaccccggg aggcatagaa aaaggggaaa gccacccagg agagaagata ttgagatgga    600 

tgaaattata gaagaacaag aagatgagaa tatttgtttt tctgcccggg acctttctca    660 

aattcgaaat gccatctttc accttttaaa gaatttttta aggcttctgc caaagttttc    720 

cttgaaagaa aagccacaat gtgtacagaa ttgtatagag gtctttgttt cattaactaa    780 

ttttgagcca gttcttcatg aatgtcatgt tacacaagcc agagctctta accaagcaaa    840 

atacatacca gaactggctt attatggatt gtatttgctg tgctctccca ttcatggaga    900 

aggagataag gtcatcagtt gtgttttcca tcaaatgctc agtgtaatat taatgttaga    960 

agttggtgaa ggatcccatc gtgcccccct tgctgttacc tcccaagtca tcaactgtag   1020 

aaaccaggcg gtccagttta tcagcgccct tgtggatgaa ttaaaggaga gtatattccc   1080 

agtcgtccgt atcttactgc agcacatctg tgccaaggtg gtagataaat cagagtatcg   1140 

tacttttgca gcccagtccc tagtccagct gctcagtaaa cttccttgtg gggaatacgc   1200 

tatgttcatt gcctggcttt acaaatactc ccgaagttcc aagatcccac accgggtttt   1260 

tactcttgat gttgtcttag ctctgttaga actgcctgaa agagaggtgg ataacaccct   1320 

ctccttggag catcagaagt tcttaaagca taagttcctg gtgcaggaaa ttatgtttga   1380 

tcgttgctta gacaaggcgc ctactgtccg cagcaaggca ctgtccagct ttgcacactg   1440 

tctggagttg actgttacca gtgcgtcgga gagtatcctg gagctcctga ttaacagtcc   1500 

tacgttttct gtaatagaga gtcaccctgg taccttactg agaaattcat cagctttttc   1560 

ctaccaaagg cagacatcta accgttccga accctcaggg gagatcaaca tagacagcag   1620 

tggtgaaaca gttggatctg gagaaagatg tgtcatggca atgctgagaa ggaggatcag   1680 

ggatgagaag accaacgtta ggaagtctgc actgcaggta ttagtgagta ttttgaaaca   1740 

ctgtgatgtc tcaggcatga aggaagacct gtggattctg caggaccagt gtcgggaccc   1800 

tgcagtgtct gtccggaagc aggccctcca gtctcttact gaactcctta tggctcagcc   1860 

tagatgcgtg cagatccaga aagcctggtt gcggggggtg gtcccggtgg tgatggactg   1920 

cgagagcact gtgcaggaga aggccctgga gttcctggac cagctgctgc tgcagaacat   1980 

ccggcatcac agtcattttc actctgggga cgacagccag gtcctcgcct gggcgcttct   2040 

tactctcctc accaccgaaa gccaggaact gagccgatat ttaaataagg cttttcatat   2100 

ctggtccaag aaagaaaaat tctcacccac ttttataaac aatgtaatat ctcacactgg   2160 

cacggaacat tcggcacctg cctggatgct gctctccaag attgctggct cctcacccag   2220 

gctggactac agcagaataa tacaatcttg ggagaaaatc agcagtcagc agaatcccaa   2280 

ttcaaacacc ttaggacata ttctctgtgt gattgggcat attgcaaagc atcttcctaa   2340 

gagcacccgg gacaaagtga ctgatgctgt caagtgtaag ctgaatggat ttcagtggtc   2400 

tctagaggtg atcagttcag ctgttgacgc cttgcagagg ctttgtagag catctgcaga   2460 

gacaccagca gaggagcagg aattgctgac gcaggtgtgt ggggatgtac tctccacctg   2520 

cgagcaccgc ctctccaaca tcgttctcaa ggagaatgga acagggaata tggacgaaga   2580 

cctgttggtg aagtacattt ttaccttagg ggatatagcc cagctgtgtc cagccagggt   2640 

ggagaagcgc atcttccttc tgattcagtc cgtcctggct tcgtctgctg atgctgacca   2700 

ctcaccatca tctcaaggca gcagtgaggc cccagcgtct cagccacccc cccaggtcag   2760 

aggttctgtc atgccctctg tgattagagc acatgccatc attaccttag gtaagctgtg   2820 

cttacagcac gaggatctgg caaagaagag catcccagcc ctggtgcgag agctcgaggt   2880 

gtgtgaggac gtggctgtcc gcaacaacgt catcattgta atgtgcgatc tctgcattcg   2940 

ctacaccatc atggtggaca agtatattcc caacatctcc atgtgtctga aggattccga   3000 

cccattcatc cggaagcaga cactcatctt gcttaccaat ctcttgcagg aggaatttgt   3060 

gaaatggaag ggctccctgt tcttccgatt tgtcagcact ctgatcgatt cacacccaga   3120 

cattgccagc ttcggggagt tttgcctggc tcacctgtta ctgaagagga accctgtcat   3180 

gttcttccaa cacttcattg aatgtatttt tcactttaat aactatgaga agcatgagaa   3240 

gtacaacaag ttcccccagt cagagagaga gaagcggctg ttttcattga agggaaagtc   3300 

aaacaaagag agacgaatga aaatctacaa atttcttcta gagcacttca cagatgaaca   3360 

gcgattcaac atcacttcca aaatctgcct tagtattttg gcgtgctttg ctgatggcat   3420 

cctacccctg gacctggacg ccagtgagtt actctcagac acgtttgagg tcctcagctc   3480 

aaaggagatc aagcttttgg caatgagatc taaaccagac aaagacctcc ttatggaaga   3540 

agatgacatg gccttggcaa atgtagtcat gcaggaagct cagaagaagc tcatctcaca   3600 

agttcagaag aggaatttca tagaaaatat tattccaatt atcatctccc tgaagactgt   3660 

gctggagaaa aataagatcc cagctttgcg ggaactcatg cactatctca gggaggtgat   3720 

gcaggattac cgagatgagc tcaaggactt ctttgcagtt gacaaacagc tggcatcaga   3780 

gcttgagtat gacatgaaga agtaccagga acagctggtc caggagcagg agctagcaaa   3840 

acatgcagat gtggccggga cggctggagg tgctgaggtg gcacctgtgg cacaggttgc   3900 

cctgtgttta gaaacagtgc cagttcctgc tggccaagaa aaccctgcca tgtcacctgc   3960 

cgtgagccag ccctgcacac ccagggcaag tgctggccat gtagcagtat catctcctac   4020 

acctgaaaca gggccattgc agaggttgct gcccaaagcc aggcccatgt ccctgagcac   4080 

cattgcaatc ctgaattctg tcaagaaagc cgtggagtca aagagcaggc atcggagtcg   4140 

gagcttagga gtgctgcctt tcactttaaa ttctggaagc ccagaaaaaa cgtgcagtca   4200 

ggtgtcttca tacagtttgg agcaagagtc gaatggcgag attgagcacg tgaccaagcg   4260 

ggccatcagc acccccgaga agagcatcag tgatgtcacg tttggagcag gggtcagtta   4320 

catcgggaca ccacggactc cgtcgtcagc caaagagaaa attgaaggcc ggagtcaagg   4380 

aaatgacatc ttatgtttat cactgcctga taaaccgccc ccacagcctc agcagtggaa   4440 

tgtgcggtct cccgccagga ataaagacac tccagcctgc agcaggaggt ccctccgaaa   4500 

gacccctctg aaaacagcca actaaacagc gcctcccacc agtgtccagg caggcaggag   4560 

cccttgagga agcagtctcg tgtcctccgt gtgaaggcag ctggatcact tcccgcagtc   4620 

cttgggcagc gctttgctgt ggaacacgag agctcctcct caggggcctg gcactcacct   4680 

tctattctgt atgatgtatt tggttaaaca ctgtcaaata atagagatgt gccagattta   4740 

gattttctta ccctaatctg tttaatattg taactttatt ccatttgaaa gtgtcaagcc   4800 

cattcagata agctataatc tggtctttaa ggaatacaac tttaaaactg cagctttctt   4860 

ttatataaat caagcctctg ttaacttgaa ttccttatag tacatatttt cccatctgta   4920 

atgccggaat tttgattcta atattttttc tattatttat aagtgcaaat ttttttaaaa   4980 

agtgtacagc tttcttaaag taataaaggt ttagcataaa tac                     5023 

 
           
             15  
             403  
             DNA  
             Homo sapiens  
           
            15 

ccatcacggg gaattctgct gctgttatta ccccattcaa gttgacaact gaggcaacgc     60 

agactccagt ctccaataag aaaccagtgt ttgatcttaa agcaagtttg tctcgtcccc    120 

tcaactatga accacacaaa ggaaagctaa aaccatgggg gcaatctaaa gaaaataatt    180 

atctaaatca acatgtcaac agaattaact tctacaagaa aacttacaaa caaccccatc    240 

tccagacaaa ggaagagcaa cggaagaaac gcgagcaaga acgaaaggag aagaaagcaa    300 

aggttttggg aatgcgaagg ggcctcattt tggctgaaga ttaataattt tttaacatct    360 

tgtaaatatt cctgtattct caactttttt ccttttgtaa att                      403 

 
           
             16  
             890  
             DNA  
             Homo sapiens  
             
               misc_feature  
               (1)...(890)  
               n = A,T,C or G  
             
           
            16 

agcataagcg tntcactgac caagactcca gccagaaagt ctgcacatgt gaccgtgtct     60 

gggggcaccc aaaaaggcga ggctgtgctt gggacacaca aattaaagac catcacgggg    120 

aattctgctg ctgttattac cccattcaag ttgacaactg aggcaacgca gactccagtc    180 

tccaataaga aaccagtgtt tgatcttaaa gcaagtttgt ctcgtcccct caactatgaa    240 

ccacacaaag gaaagctaaa accatggggg caatctaaag aaaataatta tctaaatcaa    300 

catgtcaaca gaattaactt ctacaagaaa acttacaaac aaccccatct ccagacaaag    360 

gaagagcaac ggaagaaacg cgagcaagaa cgaaaggaga agaaagcaaa ggttttggga    420 

atgcgaaggg gcctcatttt ggctgaagat taataatttt ttaacatctt gtaaatattc    480 

ctgtattctc aacttttttc cttttgtaaa tttttttttt tttgctgtca tccccacttt    540 

agtcacgaga tctttttctg ctaactgttc atagtctgtg gtagtgtcca tgggttcttc    600 

atgtgctatg atctctgaaa agacgttatc accttaaagc tcaaattctt tgggatggtt    660 

tttacttaag tccattaaca attcaggttt ctaacgagac ccatcctaaa attctgtttc    720 

tagattttta atgtcaagtt cccaagttyc ccctgctggt tctaatatta acagaactgc    780 

agtcttctgc tagccaatag catttacctg atggcagcta gttatgccag ctttagggag    840 

aatttgaaca ttttccagga atgggggaag ctgggaaaga aaggccacct               890 

 
           
             17  
             371  
             DNA  
             Homo sapiens  
           
            17 

ttggctcagc aggacaatat ggtgggaaat gacaaagtaa ctcctgtggc cctaggtcag     60 

gttctcttga ggaaaacaaa aaggctggaa tgatacagct cttcgtaaac caggtgcctc    120 

cagtgcctgc ggttattccc aagtccacat tttgcagaca gggccctaaa atgtctagct    180 

aggaagttcc tgagcctgtt tttttaaaat tctacacaca cacatgcaca cacacacgca    240 

cgtgtgcaca catgcggata tatacatcct caccttttct tgagattact gctcagaaga    300 

aggcacattt ggtttggtct gcttaccagg tgctgaagtg ggagcggccg caagcttawt    360 

tccttttagt g                                                         371 

 
           
             18  
             376  
             DNA  
             Homo sapiens  
           
            18 

attctttggc tcagcaggac aatatggtgg gaaatgacaa agtaactcct gtggccctag     60 

gtcaggttct cttgaggaaa acaaaaaggc tggaatgata cagctcttcg taaaccaggt    120 

gcctccagtg cctgcggtta ttcccaagtc cacattttgc agacagggcc ctaaaatgtc    180 

tagctaggaa gttcctgagc ctgttttttt aaaattctac acacacacat gcacacacac    240 

acgcacgtgt gcacacatgc ggatatatac atcctcacct tttcttgaga ttactgctca    300 

gaagaaggca catttggttt ggtctgctta ccaggtgctg aagtgggagc ggccgcaagc    360 

ttawttcctt ttagtg                                                    376 

 
           
             19  
             512  
             DNA  
             Homo sapiens  
             
               misc_feature  
               (1)...(512)  
               n = A,T,C or G  
             
           
            19 

ccatgtgata ctgtatgaac ctangtagnt tggaagaaaa agtagggttt ttgtatacta     60 

gcttttgtat ttgaattaat tatcattcca gctttttata tactatattt catttatgaa    120 

gaaattgatt ttcttttggg agncactttt aatctgtaan tttaaaatac aagtctgaat    180 

atttatagtt gattcttaac tgtgcatana cctagatata ccattatccc ttttatacct    240 

aanaagggca tgctaataat taccactgtc aaagaggcaa aggnggtgat ttttgnntat    300 

gaagttaagc ctcagnggag gctcatttgt tagtttttag cngganctaa ngntaaactc    360 

agggtnccct gagctatatg cacactcaga cctctttgct ttacccagng gcgttngtga    420 

gttgctcagc agtacaaact gcccttacct gacagagccc tgnctttgac ctgctcagcc    480 

ctgtgcgcta atcctctagt agcccaatca na                                  512 

 
           
             20  
             3410  
             DNA  
             Homo sapiens  
           
            20 

gcaccaggcg cccagtggag ccgtttggga gaattgcctg cgccacgcag cggggccgga     60 

caggcggtaa ggatctgatt aggctttcga acttgagttt gactgatgtc ttctgtgtgg    120 

tgtccgctaa atcccacagc atataggatc agtcgcattg gttataaggt ttgcttctgg    180 

ctgggtgcgg tggctcatgc ctgtaatcca acattgggag gccaaggcag gcggaccacc    240 

tgaagtcggg agcttgagtc cagccactgt ctgggtactg ccagccatcg ggcccaggtc    300 

tctggggttg tcttaccgca gtgagtacca cgcggtacta cagagaccgg ctgcccgtgt    360 

gcccggcagg tggagccgcc gcatcagcgg cctcggggaa tggaagcgga gaacgcgggc    420 

agctattccc ttcagcaagc tcaagctttt tatacgtttc catttcaaca actgatggct    480 

gaagctccta atatggcagt tgtgaatgaa cagcaaatgc cagaagaagt tccagcccca    540 

gctcctgctc aggaaccagt gcaagaggct ccaaaaggaa gaaaaagaaa acccagaaca    600 

acagaaccaa aacaaccagt ggaacccaaa aaacctgttg agtcaaaaaa atctggcaag    660 

tctgcaaaac caaaagaaaa acaagaaaaa attacagaca catttaaagt aaaaagaaaa    720 

gtagaccgtt ttaatggtgt ttcagaagct gaacttctga ccaagactct ccccgatatt    780 

ttgaccttca atctggacat tgtcattatt ggcataaacc cgggactaat ggctgcttac    840 

aaagggcatc attaccctgg acctggaaac catttttgga agtgtttgtt tatgtcaggg    900 

ctcagtgagg tccagctgaa ccatatggat gatcacactc taccagggaa gtatggtatt    960 

ggatttacca acatggtgga aaggaccacg cccggcagca aagatctctc cagtaaagaa   1020 

tttcgtgaag gaggacgtat tctagtacag aaattacaga aatatcagcc acgaatagca   1080 

gtgtttaatg gaaaatgtat ttatgaaatt tttagtaaag aagtttttgg agtaaaggtt   1140 

aagaacttgg aatttgggct tcagccccat aagattccag acacagaaac tctctgctat   1200 

gttatgccat catccagtgc aagatgtgct cagtttcctc gagcccaaga caaagttcat   1260 

tactacataa aactgaagga cttaagagat cagttgaaag gcattgaacg aaatatggac   1320 

gttcaagagg tgcaatatac atttgaccta cagcttgccc aagaggatgc aaagaagatg   1380 

gctgttaagg aagaaaaata tgatccaggt tatgaggcag catatggtgg tgcttacgga   1440 

gaaaatccat gcagcagtga accttgtggc ttctcttcaa atgggctaat tgagagcgtg   1500 

gagttaagag gagaatcagc tttcagtggc attcctaatg ggcagtggat gacccagtca   1560 

tttacagacc aaattccttc ctttagtaat cactgtggaa cacaagaaca ggaagaagaa   1620 

agccatgctt aagaatggtg cttctcagct ctgcttaaat gctgcagttt taatgcagtt   1680 

gtcaacaagt agaacctcag tttgctaact gaagtgtttt attagtattt tactctagtg   1740 

gtgtaattgt aatgtagaac agttgtgtgg tagtgtgaac cgtatgaacc taagtagttt   1800 

ggaagaaaaa gtagggtttt tgtatactag cttttgtatt tgaattaatt atcattccag   1860 

ctttttatat actatatttc atttatgaag aaattgattt tcttttggga gtcactttta   1920 

atctgtaatt ttaaaataca agtctgaata tttatagttg attcttaact gtgcataaac   1980 

ctagatatac cattatccct tttataccta agaagggcat gctaataatt accactgtca   2040 

aagaggcaaa ggtgttgatt tttgtatata agttaagcct cagtggagtc tcatttgtta   2100 

gtttttagtg gtaactaagg gtaaactcag ggttccctga gctatatgca cactcagacc   2160 

tctttgcttt accagtggtg tttgtgagtt gctcagtagt aaaaactggc ccttacctga   2220 

cagagccctg gctttgacct gctcagccct gtgtgttaat cctctagtag ccaattaact   2280 

actctggggt ggcaggttcc agagaatcga gtagaccttt tgccactcat ctgtgtttta   2340 

cttgagacat gtaaatatga tagggaagga actgaatttc tccattcata tttataacca   2400 

ttctagtttt atcttccttg gctttaagag tgtgccatgg aaagtgataa gaaatgaact   2460 

tctaggctaa gcaaaaagat gctggagata tttgatactc tcatttaaac tggtgcttta   2520 

tgtacatgag atgtactaaa ataagtaata tagaattttt cttgctaggt aaatccagta   2580 

agccaataat tttaaagatt ctttatctgc atcattgctg tttgttacta taaattaaat   2640 

gaacctcatg gaaaggttga ggtgtatacc tttgtgattt tctaatgagt tttccatggt   2700 

gctacaaata atccagacta ccaggtctgg tagatattaa agctgggtac taagaaatgt   2760 

tatttgcatc ctctcagtta ctcctgaata ttctgatttc atacgtaccc agggagcatg   2820 

ctgttttgtc aatcaatata aaatatttat gaggtctccc ccacccccag gaggttatat   2880 

gattgctctt ctctttataa taagagaaac aaattcttat tgtgaatctt aacatgcttt   2940 

ttagctgtgg ctatgatgga ttttattttt tcctaggtca agctgtgtaa aagtcattta   3000 

tgttatttaa atgatgtact gtactgctgt ttacatggac gttttgtgcg ggtgctttga   3060 

agtgccttgc atcagggatt aggagcaatt aaattatttt ttcacgggac tgtgtaaagc   3120 

atgtaactag gtattgcttt ggtatataac tattgtagct ttacaagaga ttgttttatt   3180 

tgaatgggga aaataccctt taaattatga cggacatcca ctagagatgg gtttgaggat   3240 

tttccaagcg tgtaataatg atgtttttcc taacatgaca gatgagtagt aaatgttgat   3300 

atatcctata catgacagtg tgagactttt tcattaaata atattgaaag attttaaaat   3360 

tcatttgaaa gtctgatggc ttttacaata aaagatatta agaattgtta              3410 

 
           
             21  
             627  
             DNA  
             Homo sapiens  
           
            21 

ggccaagaat tcggccgagg ggtgccgcgg ccatggagaa gcttagctcc atcaaatctc     60 

aaacaattta tgagattatt gataattctc aaggattcta cgtttgtcca gtggagcccc    120 

aaaatagaag caagatgaat attccattcc gcattggcaa tgccaaagga gatgatgctt    180 

tagaaaaaag atttcttgat aaagctcttg aactcaatat gttgtccttg aaagggcata    240 

ggtctgtggg aggcatccgg gcctctctgt ataatgctgt cacaattgaa gacgttcaga    300 

agctggccgc cttcatgaaa aaatttttgg agatgcatca gctatgaaca catcctaacc    360 

aggatatact ctgttcttga acaacataca aagtttaaag taacttgggg atggctacaa    420 

aaagttaaca cagtattttt ctcaaatgaa catgtttatt gcagattctt cttttttgaa    480 

agaacaacag caaaacatcc acaactctgt aaagctggtg ggacctaatg tcaccttaat    540 

tctgacttga actggaagca ttttaagaaa tcttgttgct tttctaacaa attcccgcgt    600 

attttgcctt tgctgctctt tttctag                                        627 

 
           
             22  
             1065  
             DNA  
             Homo sapiens  
           
            22 

ccttggctga ctcaccgccc tcgccgccgc accatggacg cccccaggca ggtggtcaac     60 

tttgggcctg gtcccgccaa gctgccgcac tcagtgttgt tagagataca aaaggaatta    120 

ttagactaca aaggagttgg cattagtgtt cttgaaatga gtcacaggtc atcagatttt    180 

gccaagatta ttaacaatac agagaatctt gtgcgggaat tgctagctgt tccagacaac    240 

tataaggtga tttttctgca aggaggtggg tgcggccagt tcagtgctgt ccccttaaac    300 

ctcattggct tgaaagcagg aaggtgtgcg gactatgtgg tgacaggagc ttggtcagct    360 

aaggccgcag aagaagccaa gaagtttggg actataaata tcgttcaccc taaacttggg    420 

agttatacaa aaattccaga tccaagcacc tggaacctca acccagatgc ctcctacgtg    480 

tattattgcg caaatgagac ggtgcatggt gtggagtttg actttatacc cgatgtcaag    540 

ggagcagtac tggtttgtga catgtcctca aacttcctgt ccaagccagt ggatgtttcc    600 

aagtttggtg tgatttttgc tggtgcccag aagaatgttg gctctgctgg ggtcaccgtg    660 

gtgattgtcc gtgatgacct gctggggttt gccctccgag agtgcccctc ggtcctggaa    720 

tacaaggtgc aggctggaaa cagctccttg tacaacacgc ctccatgttt cagcatctac    780 

gtcatgggct tggttctgga gtggattaaa aacaatggag gtgccgcggc catggagaag    840 

cttagctcca tcaaatctca aacaatttat gagattattg ataattctca aggattctac    900 

gtgtctgtgg gaggcatccg ggcctctctg tataatgctg tcacaattga agacgttcag    960 

aagctggccg ccttcatgaa aaaatttttg gagatgcatc agctatgaac acatcctaac   1020 

caggatatac tctgttcttg aacaacatac aaagtttaaa gtaac                   1065 

 
           
             23  
             578  
             DNA  
             Homo sapiens  
             
               misc_feature  
               (1)...(578)  
               n = A,T,C or G  
             
           
            23 

gcctcgggcc aagaattcgg cacgaggcca agttaaggaa cttgaagcta atgtacttgc     60 

tacagcccct gacaaaaaaa gcagaaattg ctagaagaaa acgttagtgc tttcaaaaca    120 

gaatangang ctgnggctga gaaagctggt aaagtagaag ctgaggttaa acgcttacac    180 

aataccatcg tagaaatcaa taatcataaa ctcaaggccc aacaagacaa acttgataaa    240 

ataaataagc aattagatga atgtgcttct gctattacta aagcccaagt agcaatcaag    300 

actgctgaca gaaaccttca aaaggcacaa gactctgtct tgcgtacaga gaaagaaata    360 

aaagatactg agaaagaggt ggatgaccta acagcagagc tgaaaagtct tgaggacaaa    420 

gcagcagagg tcgtaaagaa tacaaatgct gcagagcagt tcttttcggt gtttaggaat    480 

ccttaccaga gatccagaaa gaacatcgca atctgcttca agaattaaaa gttattcaag    540 

aaaatgaaca tgctcttcaa aaagatgcct tagtatta                            578 

 
           
             24  
             3799  
             DNA  
             Homo sapiens  
           
            24 

atagtaaacc agaacttcaa atcctatgct ggggagaaaa ttctgggacc tttccataag     60 

cgcttttcct gtattatcgg gccaaatggc agtggcaaat ccaatgttat tgattctatg    120 

ctttttgtgt ttggctatcg agcacaaaaa ataagatcta aaaaactctc agtattaata    180 

cataattctg atgaacacaa ggacattcag agttgtacag tagaagttca ttttcaaaag    240 

ataattgata aggaagggga tgattatgaa gtcattccta acagtaattt ctatgtatcc    300 

agaacggcct gcagagataa tacttctgtc tatcacataa gtggaaagaa aaagacattt    360 

aaggatgttg gaaatcttct tcgaagccat ggaattgact tggaccataa tagattttta    420 

attttacagg gtgaagttga acaaattgct atgatgaaac caaaaggcca gactgaacac    480 

gatgagggta tgcttgaata tttagaagat ataattggtt gtggacggct aaatgaacct    540 

attaaagtct tgtgtcaaag agttgaaata ttaaatgaac acagaggaga gaagttaaac    600 

agggtaaaga tggtggaaaa ggaaaaggat gccttagaag gagagaaaaa catagctatc    660 

gaatttctta ccttggaaaa tgaaatattt agaaaaaaga atcatgtttg tcaatattat    720 

atttatgagt tgcagaaacg aattgctgaa atggaaactc aaaaggaaaa aattcatgaa    780 

gataccaaag aaattaatga gaagagcaat atactatcaa atgaaatgaa agctaagaat    840 

aaagatgtaa aagatacaga aaagaaactg aataaaatta caaaatttat tgaggagaat    900 

aaagaaaaat ttacacacgt agatttggaa gatgttcaag ttagagaaaa gttaaaacat    960 

gccacgagta aagccaaaaa actggagaaa caacttcaaa aagataaaga aaaggttgaa   1020 

gaatttaaaa gtatacctgc caagagtaac aatatcatta atgaaacaac aaccagaaac   1080 

aatgccctcg agaaggaaaa agagaaagaa gaaaaaaaat taaaggaagt tatggatagc   1140 

cttaaacagg aaacacaagg gcttcagaaa gaaaaagaaa gtcgagagaa agaacttatg   1200 

ggtttcagca aatcggtaaa tgaagcacgt tcaaagatgg atgtagccca gtcagaactt   1260 

gatatctatc tcagtcgtca taatactgca gtgtctcaat taactaaggc taaggaagct   1320 

ctaattgcag cttctgagac tctcaaagaa aggaaagctg caatcagaga tatagaagga   1380 

aaactccctc aaactgaaca agaattaaag gagaaagaaa aagaacttca aaaacttaca   1440 

caagaagaaa caaactttaa aagtttggtt catgatctct ttcaaaaagt tgaagaagca   1500 

aagagctcat tagcaatgaa ttcgagtagg gggaaagtcc ttgatgcaat aattcaagaa   1560 

aaaaaatctg gcaggattcc aggaatatat ggaagattgg gggacttagg agccattgat   1620 

gaaaaatacg acgtggctat atcatcctgt tgtcatgcac tggactacat tgttgttgat   1680 

tctattgata tagcccaaga atgtgtaaac ttccttaaaa gacaaaatat tggagttgca   1740 

acctttatag gtttagataa gatggctgta tgggcgaaaa agatgaccga aattcaaact   1800 

cctgaaaata ctcctcgttt atttgattta gtaaaagtaa aagatgagaa aattcgccaa   1860 

gctttttatt ttgctttacg agatacctta gtagctgaca acttggatca agccacaaga   1920 

gtagcatatc aaaaagatag aagatggaga gtggtaactt tacagggaca aatcatagaa   1980 

cagtcaggta caatgactgg tggtggaagc aaagtaatga aaggaagaat gggttcctca   2040 

cttgttattg aaatctctga agaagaggta aacaaaatgg aatcacagtt gcaaaacgac   2100 

tctaaaaaag caatgcaaat ccaagaacag aaagtacaac ttgaagaaag agtagttaag   2160 

ttacggcata gtgaacgaga aatgaggaac acactagaaa aatttactgc aagcatccag   2220 

cgtttaatag agcaagaaga atatttgaat gtccaagtta aggaacttga agctaatgta   2280 

cttgctacag cccctgacaa aaaaaagcag aaattgctag aagaaaacgt tagtgctttc   2340 

aaaacagaat atgatgctgt ggctgagaaa gctggtaaag tagaagctga ggttaaacgc   2400 

ttacacaata ccatcgtaga aatcaataat cataaactca aggcccaaca agacaaactt   2460 

gataaaataa ataagcaatt agatgaatgt gcttctgcta ttactaaagc ccaagtagca   2520 

atcaagactg ctgacagaaa ccttcaaaag gcacaagact ctgtcttgcg tacagagaaa   2580 

gaaataaaag atactgagaa agaggtggat gacctaacag cagagctgaa aagtcttgag   2640 

gacaaagcag cagaggtcgt aaagaataca aatgctgcag aggaatcctt accagagatc   2700 

cagaaagaac atcgcaatct gcttcaagaa ttaaaagtta ttcaagaaaa tgaacatgct   2760 

cttcaaaaag atgcacttag tattaagttg aaacttgaac aaatagatgg tcacattgct   2820 

gaacataatt ctaaaataaa atattggcac aaagagattt caaaaatatc actgcatcct   2880 

atagaagata atcctattga agagatttcg gttctaagcc cagaggatct tgaagcgatc   2940 

aagaatccag attctataac aaatcaaatt gcacttttgg aagcccggtg tcatgaaatg   3000 

aaaccaaacc tcggtgccat cgcagagtat aaaaagaagg aagaattgta tttgcaacgg   3060 

gtagcagaat tggacaaaat tacttatgaa agagacagtt ttagacaggc atatgaagat   3120 

cttcggaaac aaaggcttaa tgaatttatg gcaggttttt atataataac aaataaatta   3180 

aaggaaaatt accaaatgct tactttggga ggggacgccg aactcgagct tgtagacagc   3240 

ttggatcctt tctctgaagg aatcatgttc agtgttcgac cacctaagaa aagttggaaa   3300 

aagatcttca acctttcggg aggagagaaa acacttagtt cattggcttt agtatttgct   3360 

cttcaccact acaagcccac tcccctttac ttcatggatg agattgatgc agcccttgat   3420 

tttaaaaatg tgtccattgt tgcattttat atatatgaac aaacaaaaaa tgcacagttc   3480 

ataataattt ctcttcgaaa taatatgttt gagatttcgg atagacttat tggaatttac   3540 

aagacataca acataacaaa aagtgttgct gtaaatccaa aagaaattgc atctaaggga   3600 

ctttgttgaa ctttatctga agtctcaagt tgattcaggt attactgatt tttttctatt   3660 

tgtaaaggat tatgagttgt ataaaataca tactccctaa actagatcat gaaactggtt   3720 

tctgttttat gcagttgtca tttgtaaagt ctaataaaat attctctata attgcttcta   3780 

gattacaaaa atatgacaa                                                3799 

 
           
             25  
             429  
             DNA  
             Homo sapiens  
             
               misc_feature  
               (1)...(429)  
               n = A,T,C or G  
             
           
            25 

atgggaacaa agaagtattt taaaattata actactcatt ctttctttag ccttagttaa     60 

tttgagcaga agccacaaca agcaaaccac aataaattta gaattggcag aaatccacat    120 

taactcctct tcccaagttt ccacactact accatttaca gttgtaggtt tgtaatgtat    180 

aattatgtaa tgcagaaact agctttgact tgtgtaacga tgcactgtca aagtaagcaa    240 

agtaagaatt gaaattccac attcccagaa tttaacactc agctgctcct ctagtaataa    300 

gttcctgggg ataatacatt aaccaacatt ggttgaaaca tacctgagta atcatatcag    360 

gatgcatgtt aagctgataa aacaataaga tcccaaaatg cagtagctca aaaaaaaaaa    420 

aaaaaaggn                                                            429 

 
           
             26  
             788  
             DNA  
             Homo sapiens  
             
               misc_feature  
               (1)...(788)  
               n = A,T,C or G  
             
           
            26 

nccttttttt tttttttttt gagctactgc attttgggat cttattgttt tatcagctta     60 

acatgcatcc tgatatgatt actcaggtat gtttcaacca atgttggtta atgtattatc    120 

cccaggaact tattactaga ggagcagctg agtgttaaat tctgggaatg tggaatttca    180 

attcttactt tgcttacttt gacagtgcat cgttacacaa gtcaaagcta gtttctgcat    240 

tacataatta tacattacaa acctacaact gtaaatggta gtagtgtgga aacttgggaa    300 

gaggagttaa tgtggatttc tgccaattct aaatttattg tggtttgctt gttgtggctt    360 

ctgctcaaat taactaaggc taaagaaaga atgagtagtt ataattttaa aatacttctt    420 

tgttcccata tagcaccctt tacgcgctga gatgaaaaaa cactttttgt tgagactaag    480 

agcttattac tcttcccaag attctctggc aattcagatt ccccaacttc catatcagcc    540 

attttcttct aataaaggaa ctactgatat tcttgggcaa attattacct cctctggctc    600 

agttgttttg accatgggct aatgagccca gggcctgggg tttgattccc acgcatgcca    660 

attagctttg cttgcctcca ccaacccagg ctgccctatt aaagcctgcc gcctgtccga    720 

agatgccacc acacatcttg ccttatgagt cattggtcat aaaaggggcc agctaatgag    780 

tagggaaa                                                             788 

 
           
             27  
             687  
             DNA  
             Homo sapiens  
             
               misc_feature  
               (1)...(687)  
               n = A,T,C or G  
             
           
            27 

acatggtttg tgctttactc ttaaacatct ttaaagtgct attattctat atctgttgga     60 

tgagtcatta tttttgaaat gataatccta gcatgaactc tgatctatgg tgttggattc    120 

tgtttcttaa ataactttaa aattaactgt tttcccttga gatttccttc tcctatgtag    180 

gtatttgagc tattgttcta agtttacctg taagtataaa ccttgggaga atctaagtaa    240 

acatatttct aaaagcatag ttaccttcct attttctggc tcttaccttc ttggagtatt    300 

taaatgccca tttgccaaaa gcagacctga acatcaagcc tgttaattct tcaaagaatt    360 

taggtatttg tttcaccgaa atgaagtgac ttattagcca ttcagcgtat tagtattaca    420 

gaggctcttg cccagccaca tccattcatt gatttttatg gctactcttc ccagttacat    480 

tttatgcatc tgtaagcttt ccttccttag caaaattgca ttcaaaaatg tgtaaaaatg    540 

agtaaataca gaatatcact acagagactt gnatcctcan ggttaatgga tttcacattg    600 

ngaaataaac agcaaanggt cttaagtttt caagtgaaaa ctttttgggt aatcacaaaa    660 

atacctggac acataccacg ctttaaa                                        687 

 
           
             28  
             1529  
             DNA  
             Homo sapiens  
             
               misc_feature  
               (1)...(1529)  
               n = A,T,C or G  
             
           
            28 

gagatcatcg atttaggtgg ctgcntaagt attactgatg tgtccttaca tgcattagga     60 

aaaaactrcm cmttwtwgca gtgtgtcgac ttttcagcta ctcaggtatc tgacagtggt    120 

gtgattgcac ttgttagtgg accttgtgcg aagaaattag aggagattca tatgggacat    180 

tgtgtaaatc tgactgatgg ggctgtcgaa gctgtcctta cttactgtcc tcaaatacgt    240 

atattactct tccatggatg ccccttgata acagatcatt cccgagaagt gttggagcaa    300 

ttagtaggcc caaacaaact aaagcaagtg acatggactg tttattgatg cttttttgaa    360 

gatgatcaat gctaggaaag cttatcaaaa ctactttccc aggaaaccat ctatagagat    420 

ttgcattcta cttaatgtta acactatttt taattatttt attgtcttaa gttataactc    480 

tcagagaatt agctaagtct tggtatatac atggtttgtg ctttactctt aaacatcttt    540 

aaagtgctat tattctawaw mtgttggatg agtcattatt tttgaaatga taatcctagc    600 

atgaactctg atctatggtg ttggattctg tttcttaaat aactttaaaa ttaactgttt    660 

tcccttgaga tttccttctc ctatgtaggt atttgagcta ttgttctaag tttacctgta    720 

agtataaacc ttgggagaat ctaagtaaac atatttctaa aagcatagtt accttcctat    780 

tttctggctc ttaccttctt ggagtattta aatgcccatt tgccaaaagc agacctgaac    840 

atcaagcctg gttaattctt caaagaattt aggkgattkg tttcmccgga aatgragtga    900 

cttattagcc attcagcggt attagkawta cagaggctct tgcccagcca catccantyc    960 

attgattttt awggctactc ttcccagtta cattttatgc atctgtaagc tttccttcct   1020 

tagcaaaatt gcattcaaaa atgtgtaaaa atgagtaaat acagaatatc actacagaga   1080 

cttgtatcct caggtttatt gatttcacat tgtgaaataa acagcaaagg tcttagtttt   1140 

caagtgaaaa ctttttggta atcacaaaat tacctgacac ataccacgct ttaaaccaac   1200 

ccccaaattt agcatattca ttttgccatg agccagtctt gagattttct taaaagattt   1260 

cttattttgc ctctgatgta gtgaaaaacg gggtaagtat gctaactttc ttgtatatgt   1320 

tggggggtac ttattcaact ccatttcttg tccttacaag atttataaat gtggtatgtt   1380 

tatagtgtgg atatatatgt tgccactgca aaggtggtgc atatgtatat atgtgcaaaa   1440 

tgggtaaggc ctgttctaac tatgaaattt ttctaaagac aaattcaata aaatttaata   1500 

ctgaatattt aamcaagtca aaaaaaaaa                                     1529 

 
           
             29  
             697  
             DNA  
             Homo sapiens  
             
               misc_feature  
               (1)...(697)  
               n = A,T,C or G  
             
           
            29 

aaaaaagaaa gaaagacaag aaaaagaaaa aaaaaagaaa cacctttgtc tttgtacacg     60 

tcacgngggc tcccaggaaa atgttccttc tctttttgtt ggcatgggca ctgtgggatc    120 

tggngcattc cggtcgacac tctcgtttat ttggactgta agtctgacct ctatgaataa    180 

ttacttcagc ccctgattgc tcccgtgcca agctccttgg ccaaactttc accttagctt    240 

ctggtaagtc ttgggccaag ctaagcagca tctatcaatc atcccttcag ctcctgattg    300 

gtcctgggcc aaaggcctgg gccaagctga gccacacgtt tttcaagaca gcctgtgaac    360 

taggcacatt tccttccctt cccagtcctt aaaaaccctg gacccagcct cgtagagggc    420 

accactttca gacacctatc tctgctggca aagagctttc ttctcttgct tcttaaactt    480 

tcactccaac ctcacctttg ngtttacact ccttaatctc cttagaggta gaacaaagaa    540 

ctctggatgg tatctcagac tacgagagac tggtacatct tggngcactg ctgagactat    600 

gacacttggg ttctttgagg ttggactaaa tattttacat ggagggaaat aatacaggct    660 

ttcnttttga ctggcntaat ttacttaacn aaaaagg                             697 

 
           
             30  
             1165  
             DNA  
             Homo sapiens  
             
               misc_feature  
               (1)...(1165)  
               n = A,T,C or G  
             
           
            30 

aatgctaagt ccaaagtggt taagtgacct gcccaagctc tacaatgccc tcctgaactc     60 

ggatgtcttc atttcctgtg ccagactctt aaaaaaaata aaaataaata aaaaaagaaa    120 

gtacatctaa aaaagaaaga aagacaagaa aaagaaaaaa aaaagaaaca cctttgtctt    180 

tgtacagtca gtgggctccc aggaaaatgt tccttctctt tttgttggca tgggcactgt    240 

gggatctggt gcattccggt cgacactctc gtttatttgg actgtaagtc tgacctctat    300 

gaataattac ttcagcccct gattgctccc gtgccaagct ccttggccaa actttcacct    360 

tagcttctgr taagtcttgg gccaagctaa gcagcatcta tcaatcatcc cttcagctcc    420 

tgattgrtcc ygggccaaag gcctgggcca aagctgagcc acacgttttt caagacagcc    480 

tgtgaactag gcacatatcc ttcccttccc agtccataaa aaccctggac ccagcctcgt    540 

agaggcacca ctttcagaca cctatctctg ctggcaaaga gctttcttct cttgcttctt    600 

aaactttcac tccaacctca cctttgtgtt yacrctcctt aatctcctta gaggtagaac    660 

aaagaactct ggatgttatc tcagactacg agagactgtt acatcttggt gcactgctga    720 

gactaygaca cttggtttct ttgagtttga ctaaatattt tacatgagtg taattawtac    780 

agctttcctt tttgactgtc ttattttact taacagaatg ttttgaagga tttgtccyta    840 

ttgttagtac ttttcaagat ttccttattt ttaaggstgr atgctatccc acgtggattg    900 

tacgtgccct gtttgctgaa tctactcatc cttaagggta catttgcttc caggtaacat    960 

gtttgtgact aatactacaa atgtgcatat atctattcca tgttctgctt tggtctgttt   1020 

ggggatattt ttccatacac tggattcagt accatggtgg taatcccctt gctnttggtt   1080 

gncctcaatc cgggtggatg gnacggtccc ccccaaaatt aattggccca cggaccaagg   1140 

tggtcaanga aggcctcnac cccct                                         1165 

 
           
             31  
             557  
             DNA  
             Homo sapiens  
             
               misc_feature  
               (1)...(557)  
               n = A,T,C or G  
             
           
            31 

cgcttagggc cctcgcgggg ggcttgtggg tcctcctccc cctcccactg acaactgccc     60 

caactgctct tcccgccccg gtcacagtga aaatgtagac ggggtcgttg tccgtacgac    120 

tgtgcgccag ggctcgggga ggggcgccct ccgcgtgagc gcccccctgg gaatattgaa    180 

cataatcacc tctcattcca gactatgtta ggtcttaatg gtgggaggac gcccgagtgc    240 

tcggcccgtt tcaccccgag gaggaaggac actgggtcat gacgccatca gagggcgcca    300 

gagcagggac cggacgcgag ttggagatgt tggactcgct gttggccttg ggcggctggt    360 

gctgcttcgg gattccgtgg agtgggaggg gcgcagtctc ttgaaggcgc ctgtccaaga    420 

aagagagaga agccagagat agcctgatcc tgccttncag ttcagttctg aaaaacagca    480 

ggctcttctg cggnctaggc canggcaggc taccagccac atcttctatg agccagatgc    540 

ttatgatgac ctggacc                                                   557 

 
           
             32  
             527  
             DNA  
             Homo sapiens  
             
               misc_feature  
               (1)...(527)  
               n = A,T,C or G  
             
           
            32 

atccagggag aggagtctat ctcctcaagn ttgacaactc ctactctttg tggcggncaa     60 

aatcagtcta ctacagagtc tattatacta gataaaaatg tnggtacaaa gtctggagtc    120 

tagggttggg cagaagatga catttaattt ggaaatttct ttttactttt gtggagcatt    180 

agagtcacag tttaccttat tgatattggt ctgatggntt gtgaactctt gctgggaatc    240 

aaaatttcct tgagactctt tagcattcat actttggggn taaaggagat tnctcagact    300 

catccagccc ttgggtgctg accagcagag tcactagngg atgctgaagt tacatgagct    360 

acatgttaaa tatttaaagt ctccaaaata aaacacccca acgttgacct tacccggctt    420 

gatggttagc ccctttgctg gctgctccat gtgccttatg agagcccgta agttacaggt    480 

gtcctctaat ttgaaatcca taagntaaca ngtctatatc agntgcn                  527 

 
           
             33  
             934  
             DNA  
             Homo sapiens  
           
            33 

gtaggccagc gatgacgacg aggaggaaga aggaaacatc ggttgtgaag agaaagccaa     60 

aaagaatgcc aacaagcctt tgctggatga gattgtgcct gtgtccgacg ggactgtcat    120 

gaggatgtgt atgctggcag ccatcaatat ccaagggaga ggagtctatc tcctcaagtt    180 

tgacaactcc tactctttgt ggcggtcaaa atcagtctac tacrgagtct attatactag    240 

ataaaaatgt tgttacaaag tctggagtct wgggttgggc agaagatgac atttaatttg    300 

gaaatttctt tttacttttg tggagcatta gagtcacagt ttaccttatt gatattggtc    360 

tgatggtttg tgaactcttg ctgggaatca aaatttcctt gagactcttt agcattcata    420 

ctttggggtt aaaggagatt cctcagactc atccagccct tgggtgctga ccagcagagt    480 

cactagtgga tgctgaagtt acatgagcta catgttaaat atttaaagtc tccaaaataa    540 

aacaccccaa cgttgacctt acccggctga tggttagccc cttgctgcct gctccatgtg    600 

tcttatgaga gcccgtagtt acagtgtcct ctaatttgaa atccataagt taacaagtct    660 

atatcaggtg cagctggctt tgattaaagg ccatttttaa aacttaaaaa ctcaacacct    720 

cacagattat aatagaaaaa mgaaatgggc ctcagtttga tctccgttca gaatgaccca    780 

gattgtttct gctttggggt gcagctgttt aagttcagag ttatattaca gagaattatt    840 

ttyctggaga taatctttaa acctagaatg kttcaaaacc waattggata attggaagta    900 

tccaagatac gtagaacacc cccggagaat tttc                                934 

 
           
             34  
             758  
             DNA  
             Homo sapiens  
             
               misc_feature  
               (1)...(758)  
               n = A,T,C or G  
             
           
            34 

ggctttatag cccatcctca ttgcttactg ccacccctca gctggggtcc aaggcagtac     60 

tattcagttt attcaccaga cctgcctcca gacatctact tctttcaaaa attagtgttt    120 

tccatcaagg agcatgttcc agagcatttc ccagagatgt cccaaagaac actgtccggt    180 

gctgtggcgt acagtggcaa cagcattaga ctaagtggaa catcccagca ggctgcttta    240 

gaatccgctc atttgactag atacgatgta attggctgtc tttaaaaaac gcgcacacac    300 

acacaatctg ataggcatat ctcatgccca ttcaatatgg aatgttcttc gcttgctgaa    360 

tttaagcctg tattttaagg ttttgtggtt cctcggccac aatgggtgat gtcactgata    420 

gaacgaagct gagtttccaa gggtttgggg ctgtgcaaga gtaaacacta gagcttgagt    480 

tgttatccag ctggcaagca cggaagtctt tgaagaatgt aatgtaaaaa gggaaaagaa    540 

tgtaaagctt tttgtaccaa atgagagttg gagcccagcc aacaaatgct tttccctgtg    600 

taaaagtctc tctggaaggg acattccatc tccatggtgc actctgaggg gcactgtcaa    660 

ctagagattg gccccatcca ggtgggagga acccctttgg gatggngagt atncaatctg    720 

ctgngcattt tgacaggatc tctgaatggc taggtaat                            758 

 
           
             35  
             1534  
             DNA  
             Homo sapiens  
             
               misc_feature  
               (1)...(1534)  
               n = A,T,C or G  
             
           
            35 

ngaggtaaaa ggcaaggcag catttaataa gtacctgttg tatcctttta agtgtttgtt     60 

gtggtaatcc tcacaaagac cgggactgat ggaaactcct tgctattaaa ctttttttct    120 

tgaggaattt tgcttttcaa gtgcatatac actattaata ttttttaccc aagaggagca    180 

ttctaagcta atttatgcag tgtgactgta ttaagcatta agcttccttc agagctggcc    240 

tatcggagat gctactgccc tctctacaga tgtgtctgaa atgcctgccc aaggatggcc    300 

cttagccagt taacagcttt atagcccatc ctcattgctt actgccaccc ctcagctggg    360 

gtccaaggca gtactattca gtttattcac cagacctgcc tccagacatc tacttctttc    420 

aaaaattagt gttttccatc aaggagcatg ttccagagca tttcccagag atgtcccaaa    480 

gaacactgtc cggtgctgtg gcgtacagtg gcaacagcat tagactaagt ggaacatccc    540 

agcaggctgc tttagaatcc gctcatttga ctagatacga tgtaattggc tgtctttaaa    600 

aaacgcggca cacacacaca atctgatagg gcatatctca tgcccattca atatggaatg    660 

ttcttcgctt gctgaattta agcctgtatt ttaaggtttt gtggttcctc ggccacaatg    720 

gggtgatgtc actgatagaa cgaagctgag tttccaaggg tttggggctg tgcaaggagt    780 

aaacactaga gcttgagttg ttatccagct ggcaagcacg gaagtctttg aagaatgtaa    840 

tgtaaaaagg gaaaagaatg taaagctttt tgtaccaaat gagagttgga gcccagccaa    900 

caaatgcttt tccctgtgta aaagtctctc tggaagggac attccatctc catggtgcac    960 

tctgaggggc actgtcaact agagattggc cccatccagg tgggaggaac ccctttggrr   1020 

tggtgagtat ccaatctgct gtgcatttga caggatctct gaatggctag gtaatggatc   1080 

ccaagcaggc tcacaaattt aaatgagggc tttgtgtgca gaaagaggaa taagtacaga   1140 

ttattttcct accactagat ttttggggag agtcaccatg gaatgttgac aattacttaa   1200 

aatattttaa gctcccttgc tgaattcctg tcctgtccct gaggaatcag atggtcatac   1260 

agccataggc acccacccga aatttcccta ggagttggag taatgctaga attgaagacc   1320 

ttctgagtaa agggcttctc tgccttctca gaggcaggag aattttgcac tggttgtgtt   1380 

aaatgtataa aaagctatat gttcaccagt ttactcattt ccaatgtgta gatgaataaa   1440 

atgtagtgta caaattattt gaaaatccca gaaggaaggt acttttcaaa tacagtattt   1500 

tttttaacaa ataaacttac gatttttaca gcaa                               1534 

 
           
             36  
             125  
             PRT  
             Homo sapiens  
             
               variant  
               (1)...(125)  
               Xaa = Any amino acid  
             
           
            36 

Leu Ser Ser Arg Gly Met Lys Ala Val Leu Leu Ala Asp Thr Glu Ile 
                  5                  10                  15 

Asp Leu Phe Ser Thr Asp Ile Pro Pro Thr Asn Ala Val Asp Phe Thr 
             20                  25                  30 

Gly Arg Cys Tyr Phe Thr Lys Ile Cys Lys Cys Lys Leu Lys Asp Ile 
         35                  40                  45 

Ala Cys Leu Lys Cys Gly Asn Ile Val Xaa Tyr His Val Ile Val Pro 
     50                  55                  60 

Cys Ser Ser Cys Leu Leu Ser Cys Asn Asn Arg His Phe Trp Met Phe 
 65                  70                  75                  80 

His Ser Gln Ala Val Tyr Asp Ile Asn Arg Leu Asp Ser Thr Gly Val 
                 85                  90                  95 

Asn Val Leu Leu Arg Gly Asn Leu Pro Glu Ile Glu Glu Ser Thr Asp 
            100                 105                 110 

Glu Asp Val Leu Asn Ile Ser Ala Glu Glu Cys Ile Arg 
        115                 120                 125 

 
           
             37  
             448  
             PRT  
             Homo sapiens  
             
               VARIANT  
               (1)...(448)  
               Xaa = any amino acid  
             
           
            37 

Met Ser Arg Arg Pro Cys Ser Cys Ala Leu Arg Pro Pro Arg Cys Ser 
                  5                  10                  15 

Cys Ser Ala Ser Pro Ser Ala Val Thr Ala Ala Gly Arg Pro Arg Pro 
             20                  25                  30 

Ser Asp Ser Cys Lys Glu Glu Ser Ser Thr Leu Ser Val Lys Met Lys 
         35                  40                  45 

Cys Asp Phe Asn Cys Asn His Val His Ser Gly Leu Lys Leu Val Lys 
     50                  55                  60 

Pro Asp Asp Ile Gly Arg Leu Val Ser Tyr Thr Pro Ala Tyr Leu Glu 
 65                  70                  75                  80 

Gly Ser Cys Lys Asp Cys Ile Lys Asp Tyr Glu Arg Leu Ser Cys Ile 
                 85                  90                  95 

Gly Ser Pro Ile Val Ser Pro Arg Ile Val Gln Leu Glu Thr Glu Ser 
            100                 105                 110 

Lys Arg Leu His Asn Lys Glu Asn Gln His Val Gln Gln Thr Leu Asn 
        115                 120                 125 

Ser Thr Asn Glu Ile Glu Ala Leu Glu Thr Ser Arg Leu Tyr Glu Asp 
    130                 135                 140 

Ser Gly Tyr Ser Ser Phe Ser Leu Gln Ser Gly Leu Ser Glu His Glu 
145                 150                 155                 160 

Glu Gly Ser Leu Leu Glu Glu Asn Phe Gly Asp Ser Leu Gln Ser Cys 
                165                 170                 175 

Leu Leu Gln Ile Gln Ser Pro Asp Gln Tyr Pro Asn Lys Asn Leu Leu 
            180                 185                 190 

Pro Val Leu His Phe Glu Lys Val Val Cys Ser Thr Leu Lys Lys Asn 
        195                 200                 205 

Ala Lys Arg Asn Pro Lys Val Asp Arg Glu Met Leu Lys Glu Ile Ile 
    210                 215                 220 

Ala Arg Gly Asn Phe Arg Leu Gln Asn Ile Ile Gly Arg Lys Met Gly 
225                 230                 235                 240 

Leu Glu Cys Val Asp Ile Leu Ser Glu Leu Phe Arg Arg Gly Leu Arg 
                245                 250                 255 

His Val Leu Ala Thr Ile Leu Ala Gln Leu Ser Asp Met Asp Leu Ile 
            260                 265                 270 

Asn Val Ser Lys Val Ser Thr Thr Trp Lys Lys Ile Leu Glu Asp Asp 
        275                 280                 285 

Lys Gly Ala Phe Gln Leu Tyr Ser Lys Ala Ile Gln Arg Val Thr Glu 
    290                 295                 300 

Asn Asn Asn Lys Phe Ser Pro His Ala Ser Thr Arg Glu Tyr Val Met 
305                 310                 315                 320 

Phe Arg Thr Pro Leu Ala Ser Val Gln Lys Ser Ala Ala Gln Thr Ser 
                325                 330                 335 

Leu Lys Lys Asp Ala Gln Thr Lys Leu Ser Asn Gln Gly Asp Gln Lys 
            340                 345                 350 

Gly Ser Thr Tyr Ser Arg His Asn Glu Phe Ser Glu Val Ala Lys Thr 
        355                 360                 365 

Leu Lys Lys Asn Glu Ser Leu Lys Ala Cys Ile Arg Cys Asn Ser Pro 
    370                 375                 380 

Ala Lys Tyr Asp Cys Tyr Leu Gln Arg Ala Thr Cys Lys Arg Glu Gly 
385                 390                 395                 400 

Cys Gly Phe Asp Tyr Cys Thr Lys Cys Leu Cys Asn Tyr His Thr Thr 
                405                 410                 415 

Lys Asp Cys Ser Asp Gly Lys Leu Leu Lys Ala Ser Cys Lys Ile Gly 
            420                 425                 430 

Pro Leu Pro Gly Thr Lys Lys Ser Lys Lys Asn Leu Arg Arg Leu Xaa 
        435                 440                 445 

 
           
             38  
             1050  
             PRT  
             Homo sapiens  
           
            38 

Met Ala Ala Val Lys Lys Glu Gly Gly Ala Leu Ser Glu Ala Met Ser 
                  5                  10                  15 

Leu Glu Gly Asp Glu Trp Glu Leu Ser Lys Glu Asn Val Gln Pro Leu 
             20                  25                  30 

Arg Gln Gly Arg Ile Met Ser Thr Leu Gln Gly Ala Leu Ala Gln Glu 
         35                  40                  45 

Ser Ala Cys Asn Asn Thr Leu Gln Gln Gln Lys Arg Ala Phe Glu Tyr 
     50                  55                  60 

Glu Ile Arg Phe Tyr Thr Gly Asn Asp Pro Leu Asp Val Trp Asp Arg 
 65                  70                  75                  80 

Tyr Ile Ser Trp Thr Glu Gln Asn Tyr Pro Gln Gly Gly Lys Glu Ser 
                 85                  90                  95 

Asn Met Ser Thr Leu Leu Glu Arg Ala Val Glu Ala Leu Gln Gly Glu 
            100                 105                 110 

Lys Arg Tyr Tyr Ser Asp Pro Arg Phe Leu Asn Leu Trp Leu Lys Leu 
        115                 120                 125 

Gly Arg Leu Cys Asn Glu Pro Leu Asp Met Tyr Ser Tyr Leu His Asn 
    130                 135                 140 

Gln Gly Ile Gly Val Ser Leu Ala Gln Phe Tyr Ile Ser Trp Ala Glu 
145                 150                 155                 160 

Glu Tyr Glu Ala Arg Glu Asn Phe Arg Lys Ala Asp Ala Ile Phe Gln 
                165                 170                 175 

Glu Gly Ile Gln Gln Lys Ala Glu Pro Leu Glu Arg Leu Gln Ser Gln 
            180                 185                 190 

His Arg Gln Phe Gln Ala Arg Val Ser Arg Gln Thr Leu Leu Ala Leu 
        195                 200                 205 

Glu Lys Glu Glu Glu Glu Glu Val Phe Glu Ser Ser Val Pro Gln Arg 
    210                 215                 220 

Ser Thr Leu Ala Glu Leu Lys Ser Lys Gly Lys Lys Thr Ala Arg Ala 
225                 230                 235                 240 

Pro Ile Ile Arg Val Gly Gly Ala Leu Lys Ala Pro Ser Gln Asn Arg 
                245                 250                 255 

Gly Leu Gln Asn Pro Phe Pro Gln Gln Met Gln Asn Asn Ser Arg Ile 
            260                 265                 270 

Thr Val Phe Asp Glu Asn Ala Asp Glu Ala Ser Thr Ala Glu Leu Ser 
        275                 280                 285 

Lys Pro Thr Val Gln Pro Trp Ile Ala Pro Pro Met Pro Arg Ala Lys 
    290                 295                 300 

Glu Asn Glu Leu Gln Ala Gly Pro Trp Asn Thr Gly Arg Ser Leu Glu 
305                 310                 315                 320 

His Arg Pro Arg Gly Asn Thr Ala Ser Leu Ile Ala Val Pro Ala Val 
                325                 330                 335 

Leu Pro Ser Phe Thr Pro Tyr Val Glu Glu Thr Ala Gln Gln Pro Val 
            340                 345                 350 

Met Thr Pro Cys Lys Ile Glu Pro Ser Ile Asn His Ile Leu Ser Thr 
        355                 360                 365 

Arg Lys Pro Gly Lys Glu Glu Gly Asp Pro Leu Gln Arg Val Gln Ser 
    370                 375                 380 

His Gln Gln Ala Ser Glu Glu Lys Lys Glu Lys Met Met Tyr Cys Lys 
385                 390                 395                 400 

Glu Lys Ile Tyr Ala Gly Val Gly Glu Phe Ser Phe Glu Glu Ile Arg 
                405                 410                 415 

Ala Glu Val Phe Arg Lys Lys Leu Lys Glu Gln Arg Glu Ala Glu Leu 
            420                 425                 430 

Leu Thr Ser Ala Glu Lys Arg Ala Glu Met Gln Lys Gln Ile Glu Glu 
        435                 440                 445 

Met Glu Lys Lys Leu Lys Glu Ile Gln Thr Thr Gln Gln Glu Arg Thr 
    450                 455                 460 

Gly Asp Gln Gln Glu Glu Thr Met Pro Thr Lys Glu Thr Thr Lys Leu 
465                 470                 475                 480 

Gln Ile Ala Ser Glu Ser Gln Lys Ile Pro Gly Met Thr Leu Ser Ser 
                485                 490                 495 

Ser Val Cys Gln Val Asn Cys Cys Ala Arg Glu Thr Ser Leu Ala Glu 
            500                 505                 510 

Asn Ile Trp Gln Glu Gln Pro His Ser Lys Gly Pro Ser Val Pro Phe 
        515                 520                 525 

Ser Ile Phe Asp Glu Phe Leu Leu Ser Glu Lys Lys Asn Lys Ser Pro 
    530                 535                 540 

Pro Ala Asp Pro Pro Arg Val Leu Ala Gln Arg Arg Pro Leu Ala Val 
545                 550                 555                 560 

Leu Lys Thr Ser Glu Ser Ile Thr Ser Asn Glu Asp Val Ser Pro Asp 
                565                 570                 575 

Val Cys Asp Glu Phe Thr Gly Ile Glu Pro Leu Ser Glu Asp Ala Ile 
            580                 585                 590 

Ile Thr Gly Phe Arg Asn Val Thr Ile Cys Pro Asn Pro Glu Asp Thr 
        595                 600                 605 

Cys Asp Phe Ala Arg Ala Ala Arg Phe Val Ser Thr Pro Phe His Glu 
    610                 615                 620 

Ile Met Ser Leu Lys Asp Leu Pro Ser Asp Pro Glu Arg Leu Leu Pro 
625                 630                 635                 640 

Glu Glu Asp Leu Asp Val Lys Thr Ser Glu Asp Gln Gln Thr Ala Cys 
                645                 650                 655 

Gly Thr Ile Tyr Ser Gln Thr Leu Ser Ile Lys Lys Leu Ser Pro Ile 
            660                 665                 670 

Ile Glu Asp Ser Arg Glu Ala Thr His Ser Ser Gly Phe Ser Gly Ser 
        675                 680                 685 

Ser Ala Ser Val Ala Ser Thr Ser Ser Ile Lys Cys Leu Gln Ile Pro 
    690                 695                 700 

Glu Lys Leu Glu Leu Thr Asn Glu Thr Ser Glu Asn Pro Thr Gln Ser 
705                 710                 715                 720 

Pro Trp Cys Ser Gln Tyr Arg Arg Gln Leu Leu Lys Ser Leu Pro Glu 
                725                 730                 735 

Leu Ser Ala Ser Ala Glu Leu Cys Ile Glu Asp Arg Pro Met Pro Lys 
            740                 745                 750 

Leu Glu Ile Glu Lys Glu Ile Glu Leu Gly Asn Glu Asp Tyr Cys Ile 
        755                 760                 765 

Lys Arg Glu Tyr Leu Ile Cys Glu Asp Tyr Lys Leu Phe Trp Val Ala 
    770                 775                 780 

Pro Arg Asn Phe Ala Glu Leu Thr Val Ile Lys Val Ser Ser Gln Pro 
785                 790                 795                 800 

Val Pro Trp Asp Phe Tyr Ile Asn Leu Lys Leu Lys Glu Arg Leu Asn 
                805                 810                 815 

Glu Asp Phe Asp His Phe Cys Ser Cys Tyr Gln Tyr Gln Asp Gly Cys 
            820                 825                 830 

Ile Val Trp His Gln Tyr Ile Asn Cys Phe Thr Leu Gln Asp Leu Leu 
        835                 840                 845 

Gln His Ser Glu Tyr Ile Thr His Glu Ile Thr Val Leu Ile Ile Tyr 
    850                 855                 860 

Asn Leu Leu Thr Ile Val Glu Met Leu His Lys Ala Glu Ile Val His 
865                 870                 875                 880 

Gly Asp Leu Ser Pro Arg Cys Leu Ile Leu Arg Asn Arg Ile His Asp 
                885                 890                 895 

Pro Tyr Asp Cys Asn Lys Asn Asn Gln Ala Leu Lys Ile Val Asp Phe 
            900                 905                 910 

Ser Tyr Ser Val Asp Leu Arg Val Gln Leu Asp Val Phe Thr Leu Ser 
        915                 920                 925 

Gly Phe Arg Thr Val Gln Ile Leu Glu Gly Gln Lys Ile Leu Ala Asn 
    930                 935                 940 

Cys Ser Ser Pro Tyr Gln Val Asp Leu Phe Gly Ile Ala Asp Leu Ala 
945                 950                 955                 960 

His Leu Leu Leu Phe Lys Glu His Leu Gln Val Phe Trp Asp Gly Ser 
                965                 970                 975 

Phe Trp Lys Leu Ser Gln Asn Ile Ser Glu Leu Lys Asp Gly Glu Leu 
            980                 985                 990 

Trp Asn Lys Phe Phe Val Arg Ile Leu Asn Ala Asn Asp Glu Ala Thr 
        995                 1000                1005 

Val Ser Val Leu Gly Glu Leu Ala Ala Glu Met Asn Gly Val Phe Asp 
    1010                1015                1020 

Thr Thr Phe Gln Ser His Leu Asn Lys Ala Leu Trp Lys Val Gly Lys 
1025                1030                1035                1040 

Leu Thr Ser Pro Gly Ala Leu Leu Phe Gln 
                1045                1050 

 
           
             39  
             258  
             PRT  
             Homo sapiens  
           
            39 

Gly Lys Leu Thr Gly Ile Ser Asp Pro Val Thr Val Lys Thr Ser Gly 
                  5                  10                  15 

Ser Arg Phe Gly Ser Trp Met Thr Asp Pro Leu Ala Pro Glu Gly Asp 
             20                  25                  30 

Asn Arg Val Trp Tyr Met Asp Gly Tyr His Asn Asn Arg Phe Val Arg 
         35                  40                  45 

Glu Tyr Lys Ser Met Val Asp Phe Met Asn Thr Asp Asn Phe Thr Ser 
     50                  55                  60 

His Arg Leu Pro His Pro Trp Ser Gly Thr Gly Gln Val Val Tyr Asn 
 65                  70                  75                  80 

Gly Ser Ile Tyr Phe Asn Lys Phe Gln Ser His Ile Ile Ile Arg Phe 
                 85                  90                  95 

Asp Leu Lys Thr Glu Thr Ile Leu Lys Thr Arg Ser Leu Asp Tyr Ala 
            100                 105                 110 

Gly Tyr Asn Asn Met Tyr His Tyr Ala Trp Gly Gly His Ser Asp Ile 
        115                 120                 125 

Asp Leu Met Val Asp Glu Ser Gly Leu Trp Ala Val Tyr Ala Thr Asn 
    130                 135                 140 

Gln Asn Ala Gly Asn Ile Val Val Ser Arg Leu Asp Pro Val Ser Leu 
145                 150                 155                 160 

Gln Thr Leu Gln Thr Trp Asn Thr Ser Tyr Pro Lys Arg Ser Ala Gly 
                165                 170                 175 

Glu Ala Phe Ile Ile Cys Gly Thr Leu Tyr Val Thr Asn Gly Tyr Ser 
            180                 185                 190 

Gly Gly Thr Lys Val His Tyr Ala Tyr Gln Thr Asn Ala Ser Thr Tyr 
        195                 200                 205 

Glu Tyr Ile Asp Ile Pro Phe Gln Asn Lys Tyr Ser His Ile Ser Met 
    210                 215                 220 

Leu Asp Tyr Asn Pro Lys Asp Arg Ala Leu Tyr Ala Trp Asn Asn Gly 
225                 230                 235                 240 

His Gln Ile Leu Tyr Asn Val Thr Leu Phe His Val Ile Arg Ser Asp 
                245                 250                 255 

Glu Leu 

 
           
             40  
             324  
             PRT  
             Homo sapiens  
           
            40 

Met Asp Ala Pro Arg Gln Val Val Asn Phe Gly Pro Gly Pro Ala Lys 
                  5                  10                  15 

Leu Pro His Ser Val Leu Leu Glu Ile Gln Lys Glu Leu Leu Asp Tyr 
             20                  25                  30 

Lys Gly Val Gly Ile Ser Val Leu Glu Met Ser His Arg Ser Ser Asp 
         35                  40                  45 

Phe Ala Lys Ile Ile Asn Asn Thr Glu Asn Leu Val Arg Glu Leu Leu 
     50                  55                  60 

Ala Val Pro Asp Asn Tyr Lys Val Ile Phe Leu Gln Gly Gly Gly Cys 
 65                  70                  75                  80 

Gly Gln Phe Ser Ala Val Pro Leu Asn Leu Ile Gly Leu Lys Ala Gly 
                 85                  90                  95 

Arg Cys Ala Asp Tyr Val Val Thr Gly Ala Trp Ser Ala Lys Ala Ala 
            100                 105                 110 

Glu Glu Ala Lys Lys Phe Gly Thr Ile Asn Ile Val His Pro Lys Leu 
        115                 120                 125 

Gly Ser Tyr Thr Lys Ile Pro Asp Pro Ser Thr Trp Asn Leu Asn Pro 
    130                 135                 140 

Asp Ala Ser Tyr Val Tyr Tyr Cys Ala Asn Glu Thr Val His Gly Val 
145                 150                 155                 160 

Glu Phe Asp Phe Ile Pro Asp Val Lys Gly Ala Val Leu Val Cys Asp 
                165                 170                 175 

Met Ser Ser Asn Phe Leu Ser Lys Pro Val Asp Val Ser Lys Phe Gly 
            180                 185                 190 

Val Ile Phe Ala Gly Ala Gln Lys Asn Val Gly Ser Ala Gly Val Thr 
        195                 200                 205 

Val Val Ile Val Arg Asp Asp Leu Leu Gly Phe Ala Leu Arg Glu Cys 
    210                 215                 220 

Pro Ser Val Leu Glu Tyr Lys Val Gln Ala Gly Asn Ser Ser Leu Tyr 
225                 230                 235                 240 

Asn Thr Pro Pro Cys Phe Ser Ile Tyr Val Met Gly Leu Val Leu Glu 
                245                 250                 255 

Trp Ile Lys Asn Asn Gly Gly Ala Ala Ala Met Glu Lys Leu Ser Ser 
            260                 265                 270 

Ile Lys Ser Gln Thr Ile Tyr Glu Ile Ile Asp Asn Ser Gln Gly Phe 
        275                 280                 285 

Tyr Val Ser Val Gly Gly Ile Arg Ala Ser Leu Tyr Asn Ala Val Thr 
    290                 295                 300 

Ile Glu Asp Val Gln Lys Leu Ala Ala Phe Met Lys Lys Phe Leu Glu 
305                 310                 315                 320 

Met His Gln Leu 

 
           
             41  
             410  
             PRT  
             Homo sapiens  
           
            41 

Met Glu Ala Glu Asn Ala Gly Ser Tyr Ser Leu Gln Gln Ala Gln Ala 
                  5                  10                  15 

Phe Tyr Thr Phe Pro Phe Gln Gln Leu Met Ala Glu Ala Pro Asn Met 
             20                  25                  30 

Ala Val Val Asn Glu Gln Gln Met Pro Glu Glu Val Pro Ala Pro Ala 
         35                  40                  45 

Pro Ala Gln Glu Pro Val Gln Glu Ala Pro Lys Gly Arg Lys Arg Lys 
     50                  55                  60 

Pro Arg Thr Thr Glu Pro Lys Gln Pro Val Glu Pro Lys Lys Pro Val 
 65                  70                  75                  80 

Glu Ser Lys Lys Ser Gly Lys Ser Ala Lys Pro Lys Glu Lys Gln Glu 
                 85                  90                  95 

Lys Ile Thr Asp Thr Phe Lys Val Lys Arg Lys Val Asp Arg Phe Asn 
            100                 105                 110 

Gly Val Ser Glu Ala Glu Leu Leu Thr Lys Thr Leu Pro Asp Ile Leu 
        115                 120                 125 

Thr Phe Asn Leu Asp Ile Val Ile Ile Gly Ile Asn Pro Gly Leu Met 
    130                 135                 140 

Ala Ala Tyr Lys Gly His His Tyr Pro Gly Pro Gly Asn His Phe Trp 
145                 150                 155                 160 

Lys Cys Leu Phe Met Ser Gly Leu Ser Glu Val Gln Leu Asn His Met 
                165                 170                 175 

Asp Asp His Thr Leu Pro Gly Lys Tyr Gly Ile Gly Phe Thr Asn Met 
            180                 185                 190 

Val Glu Arg Thr Thr Pro Gly Ser Lys Asp Leu Ser Ser Lys Glu Phe 
        195                 200                 205 

Arg Glu Gly Gly Arg Ile Leu Val Gln Lys Leu Gln Lys Tyr Gln Pro 
    210                 215                 220 

Arg Ile Ala Val Phe Asn Gly Lys Cys Ile Tyr Glu Ile Phe Ser Lys 
225                 230                 235                 240 

Glu Val Phe Gly Val Lys Val Lys Asn Leu Glu Phe Gly Leu Gln Pro 
                245                 250                 255 

His Lys Ile Pro Asp Thr Glu Thr Leu Cys Tyr Val Met Pro Ser Ser 
            260                 265                 270 

Ser Ala Arg Cys Ala Gln Phe Pro Arg Ala Gln Asp Lys Val His Tyr 
        275                 280                 285 

Tyr Ile Lys Leu Lys Asp Leu Arg Asp Gln Leu Lys Gly Ile Glu Arg 
    290                 295                 300 

Asn Met Asp Val Gln Glu Val Gln Tyr Thr Phe Asp Leu Gln Leu Ala 
305                 310                 315                 320 

Gln Glu Asp Ala Lys Lys Met Ala Val Lys Glu Glu Lys Tyr Asp Pro 
                325                 330                 335 

Gly Tyr Glu Ala Ala Tyr Gly Gly Ala Tyr Gly Glu Asn Pro Cys Ser 
            340                 345                 350 

Ser Glu Pro Cys Gly Phe Ser Ser Asn Gly Leu Ile Glu Ser Val Glu 
        355                 360                 365 

Leu Arg Gly Glu Ser Ala Phe Ser Gly Ile Pro Asn Gly Gln Trp Met 
    370                 375                 380 

Thr Gln Ser Phe Thr Asp Gln Ile Pro Ser Phe Ser Asn His Cys Gly 
385                 390                 395                 400 

Thr Gln Glu Gln Glu Glu Glu Ser His Ala 
                405                 410 

 
           
             42  
             484  
             DNA  
             Homo sapiens  
           
            42 

ttcacgtaag actttttggt ttgatcatct ttgttgaggt aggactatca gttccctcta     60 

aatgtatatg ttgatttatg agtaattgtt atttattctt tatttattta tattaattat    120 

gaagattatg atattatttg attgcagatt tttttggcgc gctgccccct ccccaccctg    180 

ccactcttga cattccactg tgcgttttag aagagagcct ttttctaaag ggatctgctt    240 

aaagttttaa cttttatacc tatctgagtg aattacagac aacctatcat ttattctgct    300 

tcgagggtcc ccagggccct tgtacaaccg acagctctta cttttaaatg caatctcttt    360 

tctacataca ttattttctt aattgttagc tatttataga aagcttcaat agaactgttt    420 

caactgtata actatttact attcaaataa aatattttca aagtcaaaaa aaaaaaaaaa    480 

aaag                                                                 484 

 
           
             43  
             700  
             DNA  
             Homo sapiens  
           
            43 

ctcaccagta attccactcc catgaaactt tggtcattgt tatgcattaa gtggggctta     60 

tctttggttt ggagttcatt tgaactcttg aaccttagtt tagtgaagat gaactgtctg    120 

ttcttaggta gaaacggtgt ttatttaaaa atcagtttta aaaaatgagc taccatatgt    180 

gctgtctatt ataaatggga caccaaacaa aattttctat tacagttgtg tacttgcaaa    240 

cattttgcta tacagtactt catagatgca tacaaatgag ctcacttatt acaaagacaa    300 

acgtttaatt tgctaaatat tttaacaagt ttgttatata ttttatttaa tttaaaagaa    360 

atctcttacc aacctacata tttattacta taatttgcta tgacttcagg ttaatttatt    420 

tgtgtttgca tagtttgagc aggatgtttt gtgaagtatg tttgtattta tttgcctact    480 

ttgtacttga tgtgttttgt aatgtgcact gaatttgttt tcttttcaac tatgttaatg    540 

atcaatactg taaattgggt cttttgtaaa caaaaaggca atgatgtatg catttttttt    600 

aatttgaggt agtttgtttg tatactgttt ctccaaacac ttaatatttc ttacatcaaa    660 

gcaacaaaat tgtgttcagt gctgtacatt tggtgtatgg                          700 

 
           
             44  
             672  
             DNA  
             Homo sapiens  
             
               misc_feature  
               (1)...(672)  
               n = A,T,C or G  
             
           
            44 

tttttgttta cataattgta aggaacagta attctagaaa cactagaaga aaaargcata     60 

gcaatgtcca cagttaaaaa aaaaagkgca cattactcgg tcacaatcac agtcattact    120 

tgaaaaacta tatgtaacaa gtagataaga aatatcactg atgcctcaaa ctcattgtca    180 

aaaactgaat gacataaatt ttacatgaaa taaggcaaat tcaggaatgc acaaagaatt    240 

tgtaatccaa ccaaatctaa acaacagaaa aaagttgtat aagaagcatg aactaaagta    300 

cttctcccta aatatttaaa aaataggctt gtctcagtgc acaaagaaaa catcactcat    360 

gtgtatccca cactataaaa taagaaagaa gggtaaagta tgggggatag gagggcacag    420 

ttcattgtaa gttgcagctg catccgctga gagttcctta cattattttt agctagaact    480 

gaaaattata caaatcatat caggagatgt aatggtcttt ttggaaacta tttctgaaag    540 

aaatgaaaag aaaactacac acaagagtgc aaattttcag attgtcactt gcaacctctt    600 

aacattcagt catctacatc caggtgctgc tagagggatg cctggagaca gcagcggcaa    660 

tcaggaacga gc                                                        672 

 
           
             45  
             480  
             DNA  
             Homo sapiens  
           
            45 

tcagttccat gtatacaatt accagatgcc accgcagtgc cctgttgggg agcaaaggag     60 

aaatctgtgg accgaagcat acaaatggtg gtatcttgtc tgtttaatcc agagaagaga    120 

ctgataaatt ccgttgttac tcaagatgac tgcttcaagg gtaaaagagt gcatcgcttt    180 

agaagaagtt tggcagtatt taaatctgtt ggatcctctc agctatctag tttcatggga    240 

agttgctggt tttgaatatt aagctaaaag ttttccacta ttacagaaat tctgaatttt    300 

ggtaaatcac actgaaactt tctgtataac ttgtattatt agactctcta gttttatctt    360 

aacactgaaa ctgttcttca ttagatgttt atttagaacc tggttctgtg tttaatatat    420 

agtttaaagt aacaaataat cgagactgaa agaatgttaa gatttatctg caaggatttt    480 

 
           
             46  
             427  
             DNA  
             Homo sapiens  
             
               misc_feature  
               (1)...(427)  
               n = A,T,C or G  
             
           
            46 

tttttaaaaa taagtgtcct actattgtat tatatattga tacgaaactg ttaaagctat     60 

tttgaaaata tgagttctta gctttaatca tgaagtctga agtttgcttt cagtaattat    120 

tttaaaagtt gttttggttc attgctttat aatatttatt attgaatgcc aaacctgttc    180 

ttttttttac tgtgtccaat attctttcaa gcaaatgcaa tggctggaat ataattcaga    240 

attaactgaa acccagccag aagagggacc acctgtaaag caagtccttt caagtttcac    300 

tgcacatccc aaaccatgtt acaaaaagag caactgctat attcacatta tgatattttt    360 

ctatcttaaa tttgtcaaaa taaagtatga gtctaactat taaaaaaaaa aaaaccctck    420 

tsccaaa                                                              427 

 
           
             47  
             581  
             DNA  
             Homo sapiens  
           
            47 

tcttttgaaa aataaaggat ctaatgtctc cctaataagt cttctttcct tccaactaaa     60 

tgacctacac ggacttttat tttcttgatc aaagaggtgt ttattaagga cttctggata    120 

actatacttt tactctattt ttaaagatca caaagtaatt ttaaatgtga acaggttccc    180 

ataccatgaa tgctggcctc accttctcta tcatccacat tttgaaatgc aaagaaagct    240 

cccttgtaag ccatacttcc ttccccactc ccatcctagg atacttgccc agtgctcatt    300 

aggcatttct tattcagata gtccaaattt aggttattat gcttaatttg acacattaac    360 

taaatgccca gttttaaaat atatccatca attcacgctg aaatgtgctt ctttgtgcta    420 

tcaaatggaa tagaatacac ttatttttta aacaatccca gaatactgtg tgtagacttt    480 

tgttgtgctc aaataaatgt ttacttatct tacaaagctc aaatactgga ttgtaaccat    540 

gtgatgaagt tatctatgtt gtacctaaca tgcaaattat c                        581 

 
           
             48  
             491  
             DNA  
             Homo sapiens  
             
               misc_feature  
               (1)...(491)  
               n = A,T,C or G  
             
           
            48 

ccgggccccc cctcgagggy ttcaatggtc agatggaaca gttgaaaggc gcggtcgaaa     60 

ccctcgccat cacgatcgcg caatctggca ttctggaatt cgtcacaacg atcgtcaccg    120 

ccttgggcaa ctttgtcgat aagctcgccg aggtcagccc ggaaactctg aagtgggtca    180 

cgatcatcgg tggggtggcg gcggtgctag gtccggtggc gatcggcatc ggcgccgtgg    240 

tctctgcgct gggcgccttt ctccctgtca tcgtgcctgt tgcgagcgcc atcggcgctg    300 

tcgtttcggt catcacggcc ggtgccatcc cagccctggc cgggcttgtt gttgccctat    360 

cgcctgtgct cgtgccgctg gcggcggtgg ctgctgcagt cggcgccgtt tatctggtgt    420 

ggaagaactg ggacatgatc gggcccattc tcgccaagct ttataacgga gtgaagacgt    480 

ggctggtcga t                                                         491 

 
           
             49  
             1929  
             DNA  
             Homo sapiens  
             
               misc_feature  
               (1)...(1929)  
               n = A,T,C or G  
             
           
            49 

ttaggctagt agaggctggt gttaatcggc cgagggccgc tgtcaggttg gagtcgccga     60 

cccgttcgcg ctggcgcagc acaaatgctc gcgcatcgtg cgtgtggagt accgctgtcc    120 

cgagtgcgcc aaggtcttca gctgcccggc caacctggcc tcgcaccgcc gctggcacaa    180 

accgcggccc gcgcccgccg ccgcccgcgc gccggagcca gaagcagcag ccaggctgag    240 

gcgcgggagg cacccggcgg cggcagcgac cgggacacgc cgagccccgg cggcgtgtcc    300 

gagtcgggct ccgaggacgg gctctacgag tgccatcact gcgccaagaa gttccgccgc    360 

caggcctacc tacgcaagca cctgctggcg caccaccagg cgctgcaggc caagggcgcg    420 

ccgctagcgc ccccggccga ggacctactg gccttgtacc ccgggcccga cgagaaggcg    480 

ccccaggagg cggccggcga cggcgagggg gccggcgtgc ttgggcctga gtgcgtccgs    540 

cgagtgccac cctgtgccca gtgtgcggag agtcgttcgc cagcaaggsc gctcaggagc    600 

rccrcctgcg ccstgctgca cgccgsccag gtgttcccct gcaagtactg sctcttggca    660 

ccttctacag ctcgcccggc cttacgcggc acatcaacaa gtgccaccca tccgaaaaca    720 

gacaggtgat cctcctgcag gtgcccgtgc gcccggcctg ctagagcgcg ccctccaccc    780 

cggcccccga actgtgcctt cgcttggaga cccacaaaga gagtgcgccc tgcacgcccc    840 

gaacccgagt ccgcgctggg ggagcctcgc ccccgccccc accgggtgaa agtgtcgtct    900 

ccgcttctct cggtgtggcg tgacggtaac cccatactct ccttttgact ccttttggaa    960 

cccccacttt tacgttgtgt ccctccgcct cccccatggc gcaacaggag tcagtctctt   1020 

tctgtacaag ggagaaaagc tgtacgcgtt tgtctcgtgg ttggaagcct ccccttggcg   1080 

gggagaagct ttttttcttg ctagtattcg ctgtgttcat ggtctagaaa tgcggtctgg   1140 

tctcgcctcg cctaccaatc tctgctctct atgtatgtag cgtacgggtt gttttgggtg   1200 

aatcttgagg aataaatgcc tttatatttc acaggctgta aattgaactt cccacacgat   1260 

tagctttatt atggcttgtg aactgctgga gtctggcttt acctttttgt atgtgaacaa   1320 

atcaaattgc ttaaaaaaga gttttcttta gtatagccac aaatgccttg aactgttgtc   1380 

tgggattgtt ttgtgggggg agggaaggga gtgttccgaa gatgctgtag taactgcctc   1440 

agtgtttcac gtaagacttt ttggtttgat catctttgtt gaggtaggac tatcagttcc   1500 

ctctaaatgt atatgttgat ttatgagtaa ttgttattta ttctttattt atttatatta   1560 

attatgaaga ttatgatatt atttgattgc agattttttt ggcgcgctgc cccctcccca   1620 

ccctgccact cttgacattc cactgtgcgt tttagaagag agcctttttc taaagggatc   1680 

tgcttaaagt tttaactttt atacctatct gagtgaatta cagacaacct atcatttatt   1740 

ctgcttcgag ggtccccagg gcccttgtac aaccgacagc tcttactttt aaatgcaatc   1800 

tcttttctac atacattatt ttcttaattg ttagctattt atagaaagct tcaatagaac   1860 

tgtttcaact gtataactat ttactattca aataaaatat tttcaaagtc aaaaaaaaaa   1920 

aaaaaaaag                                                           1929 

 
           
             50  
             6183  
             DNA  
             Homo sapiens  
           
            50 

ctttttgtag ggagaagggc aggatgtttt taactgaatg tgacctcagg ggaatactag     60 

agaaaataat aaaatttctg aatggggcag cgtggagaaa tcctaagaga aatagcataa    120 

gagcattttg gaacacatcc aggaaaagat aactttcgac acacctgtag acgttcgcca    180 

ggtaaaggag tgatggaaac tctccagttc agatccagta gcttttaggg aaggaactac    240 

agttgctgac ttaagttgaa gaagcatcta tttaatgtct ggtcaaatcc tacaagaaac    300 

acagaaatct atgattaaaa agctgagcac tttgatatac tgcaaagggt agagaaggca    360 

ggacggtaga aattttctgc aagaaagaat gaatttcagg atttatcact aaataagaca    420 

aagtcattta tttagtcccc ctgacacagc agggcaaact gagttgacat acaagttacc    480 

tggagaaaaa gagagcaatt ccaggacttc ctcttcagcc taaaagaagg taccagatct    540 

gtgcactggg gcgatgtgga agagacctgc ttattgcccc tgatgtaagc tccagtaaga    600 

aaagacgtca agtacaagta ctaggaaatc actttataca tctgtttata ggaatgacct    660 

caggactttg tgttcatgtt atagatggat gcagaggctg aagataaaac gctgcgtact    720 

cgctctaaag gaaccgaggt gccaatggat tcactaatcc aggagctcag tgttgcctat    780 

gattgctcca tggcaaagaa gagaacagct gaagatcagg ctttgggggt tccagtcaac    840 

aaaaggaaat ccctgctaat gaagccccga cactacagcc caaaagcaga ctgccaagaa    900 

gaccgcagtg acaggacaga ggacgatggc cccttggaaa cacatggtca ctctaccgca    960 

gaggaaatca tgataaaacc tatggatgaa agtcttcttt caactgcaca agaaaactcc   1020 

agtaggaagg aagacagata ctcttgttat caagagctca tggtcaagtc tttaatgcac   1080 

ttggggaaat ttgaaaaaaa tgtatctgtt cagactgtaa gtgaaaattt aaatgacagt   1140 

ggcatccagt ctttaaaagc agagagcgat gaagcagacg agtgctttct gattcattct   1200 

gatgatggaa gagacaagat tgatgattct cagccaccct tctgctcctc tgatgacaat   1260 

gaaagtaact ctgaaagtgc agaaaatggc tgggacagtg gctccaactt ctcagaagaa   1320 

accaaaccac ctagagtccc aaagtatgtt ttaacagatc ataaaaaaga cctattggaa   1380 

gttcctgaaa taaaaactga aggtgacaaa tttatccctt gtgagaacag gtgtgattct   1440 

gaaacagaaa ggaaagaccc gcagaatgct ctcgcagaac ccctggatgg caatgcccag   1500 

ccctcattcc ctgacgttga ggaggaagat agcgagagcc tggcagtaat gacggaagag   1560 

ggtagtgacc tggaaaaggc caaggggaat ttaagtttgc tggagcaggc aattgctctg   1620 

caggctgagc gaggttgtgt tttccataac acctacaaag agctggatag gttcctgctg   1680 

gagcacctag caggggaaag gaggcaaacc aaagttatcg acatgggtgg aagacaaatc   1740 

tttaacaata aacattcacc aaggcctgaa aagagggaga ccaagtgccc gatccctgga   1800 

tgtgatggca cgggacacgt gacagggctc tacccgcacc accgcagcct ttcggggtgc   1860 

ccccacaaag tgcgggttcc cctggaaatt cttgccatgc atgaaaatgt gctcaagtgt   1920 

cccacgccgg gatgcacagg aaggggtcat gtgaacagca accgcaacac ccacaggagt   1980 

ctttctggtt gtccaattgc tgcagctgaa aaattggcaa tgtcccagga taaaaatcag   2040 

cttgattctc cccaaactgg gcagtgtcct gaccaggccc acaggacaag tttggtgaag   2100 

caaattgaat tcaatttccc gtcacaagcc atcacctctc ccagagccac agtgtcaaaa   2160 

gaacaagaga agtttggaaa agtaccattt gattatgcca gttttgatgc ccaagttttc   2220 

ggtaaacgcc ctctcataca aacagtgcaa ggacgaaaaa caccaccatt tcctgaatca   2280 

aagcattttc caaatccagt gaaatttcct aatcgactgc ctagtgcagg cgcccacacc   2340 

cagagccctg gccgtgccag ctcttatagc tacggtcaat gtagtgaaga cacccacata   2400 

gcagcagctg ctgccatcct gaacctttcc acccgctgca gggaagccac agacatcctc   2460 

tccaacaagc cacagagtct gcatgccaag ggagccgaaa tagaagtgga tgaaaatggc   2520 

acattggact taagcatgaa aaaaaatcga atcctggaca agtctgcacc cctaacttcc   2580 

tctaacactt ctattccaac tccttcctct tccccattca aaacaagcag cattctggtc   2640 

aatgcagcat tctatcaggc tctttgtgac caagagggct gggacactcc tatcaactat   2700 

agcaaaactc acgggaagac agaggaggag aaagagaaag acccagtgag ctctctagaa   2760 

aatttagagg aaaaaaagtt tcctggagag gcctctatac caagccctaa acccaagctt   2820 

catgcaagag atctcaaaaa ggaactaatc acctgtccaa caccaggatg tgatggaagt   2880 

ggccacgtga caggaaacta tgcatctcat cgcagtgttt ctggatgtcc tttagcagat   2940 

aagactctaa aatccctcat ggctgccaac tctcaggagc ttaagtgtcc aaccccaggc   3000 

tgcgatggct cggggcacgt gactggaaac tatgcttccc acagaagctt gtccggatgc   3060 

cctcgtgcaa ggaaaggtgg tgtcaaaatg acccctacca aggaagaaaa agaagaccct   3120 

gaactgaaat gtcctgtgat agggtgtgat ggccaaggtc acatatcagg taaatacaca   3180 

tcacaccgca cagcttctgg ctgtcctctg gctgccaaga gacagaagga gaatcctctc   3240 

aatggagcct ccctctcctg gaaactgaac aaacaagagc taccacattg tcccttgcca   3300 

ggctgcaatg ggctgggcca tgtaaataat gtttttgtca cccaccgaag cttatctgga   3360 

tgtcctctca atgcacaagt tatcaaaaag ggcaaggttt ctgaagaact catgaccatc   3420 

aagctcaaag caactggggg aatagagagt gatgaagaaa ttaggcattt ggatgaagaa   3480 

ataaaggaac tgaatgaatc caaccttaaa attgaagcag atatgatgaa acttcagacc   3540 

cagatcacat ctatggagag caacttaaag acgatagagg aggagaacaa actcatagaa   3600 

cagaacaatg aaagtctgct gaaagagctg gcaggtctaa gccaagctct catttcaagc   3660 

cttgctgaca tccagcttcc acagatggga cctatcagtg agcagaattt tgaagcatat   3720 

gtaaatacac tcacagatat gtacagcaat ctggaacggg actattcccc ggaatgcaaa   3780 

gctctactgg aaagtatcaa acaggcagtg aagggtatcc atgtgtagga tcacagcgct   3840 

gccgggcaac agaagttacc aacagcagta aactccagat ggatctgtta gaggttcatg   3900 

tactgctaag gcgtggaggt tgccgtactg catttacaat ttgcaacatt gcactaattt   3960 

tattttcccc agctgatata aaaaggaaag aaaaactatg atagacttct tggattaaaa   4020 

gcaatgcagt caattattag atcttattta ttttcatatg tttttctttt atttcttcat   4080 

tgtactcttc ttttgtaaag tatatgtaaa ataaatgtga catttttata atttatttat   4140 

tactaatcaa agagtttttt atcttttaac tgcattttga agtctgccgt atttttacaa   4200 

gtgtgtttat taatttattt tccaatagga tttaaataga aatgctattc tcaagtcatc   4260 

tttcttgctg ggttttaatg aggaaacagg aaagggtgaa ggaaatcctt gtctaaggac   4320 

tgcactatag ttgagtttga tttttattgc acacttcttc ccccaccttt cactgatttt   4380 

tgtatttata aatgaatttg cggtaaggtg agctgcacgg aaggaataag aagacaaatg   4440 

gcgcccacta gtggggaatc cgcactcaca aaagcacagg atgctggaaa acagcctgct   4500 

cagaatttgt tagcaataat taaatatagc aatcagcaaa gtattcgact tggctggacg   4560 

gttttcgtta atatgaatta tttatttgaa atgttttaaa gaaacataag cctttttagt   4620 

gatgcagatt tgtctgtttg tttttcaagt catatcagat cgttggcaac tcgtatccca   4680 

agatgaaaaa taagacttgg tgtgaccagc caggctttcc tgccatatgt tggtacaata   4740 

tacaagtgac aatattggtg tagatttgta cttagcaaat acaaacacat ccaaatgaaa   4800 

aattttgtag ataccatatc ccctgaaata gcatttatct tactgggttg actggaaagg   4860 

aatggaaaat atagtaacac atgaaaaaat gctactccaa tctgaatgat tacttcaaac   4920 

actggcacct tgggtctcac ccaccatagg aaacaagaca acattcaatt tgatagaaat   4980 

cttgccacaa aacttcaaat gctacaaaat atacacacac actcacacac acaggcatac   5040 

tcacacacag acacacacac acacacacac acagactcat ccacacttca aattgagccc   5100 

acaatcttga atttctgaac ggatcagagt ttcatagttt ctatagtaaa ggcaatgtct   5160 

atttcaggga ttgtaaagta gttaagcatt gtttcaaaag tttttttata tttatttttt   5220 

ttaaggaaaa ggtatagaca accagctaaa ctgccttttt ggtgtgcaca cacatttcat   5280 

gtgcagacgt gcctctgtgt aaatgtacac atgaacttca tgtgggctta attttctgtg   5340 

ctataaacaa aagtgtttat tttttattaa cctcatggat atttagatgg aaagtgatgg   5400 

cattcacagg cttgatgtat tccactgtta ttactgttac ctgcacaaat gaaaaacaat   5460 

actcaacagt aattccactc ccatgaaact ttggtcattg ttatgcatta agtggggctt   5520 

atctttggtt tggagttcat ttgaactctt gaaccttagt ttagtgaaga tgaactgtct   5580 

gttcttaggt agaaacggtg tttatttaaa aatcagtttt aaaaaatgag ctaccatatg   5640 

tgctgtctat tataaatggg acaccaaaca aaattttcta ttacagttgt gtacttgcaa   5700 

acattttgct atacagtact tcatagatgc atacaaatga gctcacttat tacaaagaca   5760 

aacgtttaat ttgctaaata ttttaacaag tttgttatat attttattta atttaaaaga   5820 

aatctcttac caacctacat atttattact ataatttgct atgacttcag gttaatttat   5880 

ttgtgtttgc atagtttgag caggatgttt tgtgaagtat gtttgtattt atttgcctac   5940 

tttgtacttg atgtgttttg taatgtgcac tgaatttgtt ttcttttcaa ctatgttaat   6000 

gatcaatact gtaaattggg tcttttgtaa acaaaaaggc aatgatgtat gcattttttt   6060 

taatttgagg tagtttgttt gtatactgtt tctccaaaca cttaatattt cttacatcaa   6120 

agcaacaaaa ttgtgttcag tgctgtacat ttggtgtatg gtaggaaata aaaattgata   6180 

acg                                                                 6183 

 
           
             51  
             1704  
             DNA  
             Homo sapiens  
             
               misc_feature  
               (1)...(1704)  
               n = A,T,C or G  
             
           
            51 

tccagaaaaa taaaagatat ataggagcca caagtgtctt ggggaccata taaaacaccg     60 

tgtttgggtg cctattagaa tataacgttg ggcctgctgc ctgttacgag tgtacaatgc    120 

cttctcgccg gtttgttcaa tatacccgcc cgcgccgtat ctttcgcaag gcagtttaca    180 

gccctacacc gcaggttacc cagaggtaat cgggagagct taaaataacc gttactcctg    240 

aaaaaaggta tgtaaagagc gaattttctc agtcatagtt gaataatcaa tgaagtagtc    300 

ttgcttccta atgtccttac ccattcttgg ataattcttt attagaatga atgttgagag    360 

cctgggggat cttaggatat tcttgagaaa taaatttgaa gtgccatttt gtgctaaacg    420 

taggtagaaa atggcgtttt agattttcaa aagtaaatgg ctaaaaatta agcattatac    480 

ccttcagaaa gtttataagg tttgaccatc atttttttaa cacagaaatc tgtttattaa    540 

accaaacaaa acagagaaaa ttataccagc cctcaatttt tgaattttca tttaaataag    600 

caaactctaa atccacatct taaaagatgt ttgtgcagct atgtatttcc aaaatactca    660 

tatttcaata agatttttca cattatattc accaacagta tcacaaaagt tttttttttg    720 

ttttttgttt acataattgt aaggaacagt aattctagaa acactagaag aaaaaagcat    780 

agcaatgtcc acagttacaa gaaaaagtgc acattactcg gtcacaatca cagtcattac    840 

ttgaaaaact atatgtaaca agtagataag aaatatcact gatgcctcaa actcattgtc    900 

aaaaactgaa tgacataaat tttacatgaa ataaggcaaa ttcaggaatg cacaaagaat    960 

ttgtaatcca accaaakcta aacaacagaa aaaagttgta taagaagcat gaactaaagt   1020 

acttctccct aaatatttaa aaaataggct tgtctcagtg cacaaagaaa acatcactca   1080 

tgtgtatccc acactataaa ataagaaaga agggtaaagt atgggggata ggagggcaca   1140 

gttcattgta agttgcagct gcatccgctg agagttcctt acattatttt tagctagaac   1200 

tgaaaattat acaaatcata tcaggagatg taatggtctt tttggaaact atttctgaaa   1260 

gaaatgaaaa gaaaactaca cacaagagtg caaattttca gattgtcact tgcaacctct   1320 

taacattcag tcatctacat ccaggtgctg ctagagggat gcctggagac agcagcggca   1380 

atcaggaacg agcagctcta agaaaccaag gtgtgatttt ttttcaacaa catgtcttgt   1440 

cattattaaa aaaaaaattc tgggatgaaa actgctatga taaagttgca gtgttgagtg   1500 

gggtttttga gatcagcatg agagcagaaa tgcaggcttc tcttggaagt agttcctgat   1560 

gtgacgattg aaagaacgta ggcaagggtt tttccagcat caagtgttat ttttgtagaa   1620 

agaatttgga aagaggagaa ggcaaaggga tgtggaaaag gtacttacag tagtttctca   1680 

aaacagtttt cttttaggac ctat                                          1704 

 
           
             52  
             1886  
             DNA  
             Homo sapiens  
             
               misc_feature  
               (1)...(1886)  
               n = A,T,C or G  
             
           
            52 

taaattccgt tgttactcaa gatgactgct tcaagggtaa aagagtgcat cgctttagaa     60 

gaagtttggc agtatttaaa tctgttggat cctctcagct atctagtttc atgggaagtt    120 

gctggttttg aatattaagc taaaagtttt ccactattac agaaattctg aattttggta    180 

aatcacactg aaactttctg tataacttgt attattagac tctctagttt tatcttaaca    240 

ctgaaactgt tcttcattag atgtttattt agaacctggt tctgtgttta atatatagtt    300 

taaagtaaca aataatcgag actgaaagaa tgttaagatt tatctgcaag gatttttaaa    360 

aaattgaaac ttgcatttta agtgtttaaa agcaaatact gactttcaaa aaagttttta    420 

aaacctgatt tgaaagctaa caattttgat agtctgaaca caagcatttc acttctccaa    480 

gaagtacctg tgaacagtac aatatttcag tattgagctt tgcatttatg atttatctag    540 

aaatttacct caaaagcaga atttttaaaa ctgcattttt aatcagtgga actcaatgta    600 

tagttagctt tattgaagtc ttatccaaac ccagtaaaac agattctaag caaacagtcc    660 

aatcagtgag tcataatgtt tattcaaagt attttatctt ttatctagaa nccacatatc    720 

tatgtccaat ttgatnggga tagtagttag gataactaaa attctgggcc taatttttta    780 

aagaatccaa gacaaactaa actttactgg gtatataacc ttctcaatga ggtaccattc    840 

ttttttataa aaaaaattgt tccttgaaat gctaaactta atggctgtat gtgaaatttg    900 

caaaatactg gtattaaaga acgctgcagc ttttttatgt cactcaaagg ttaatcggag    960 

tatctgaaag gaattgtttt tataaaaaca ttgaagtatt agttacttgc tataaataga   1020 

tttttatttt tgttttttag cctgttatat ttccttctgt aaaataaaat atgtccagaa   1080 

gaggcatgtt gtttctagat taggtagtgt cctcatttta tattgtgacc acacagctag   1140 

agcaccagag cccttttgct atactcacag tcttgttttc ccagcctctt ttactagtct   1200 

ttcaggaggt ttgctcttag aactggtgat gtaaagaatg gaagtagctg tatgagcagt   1260 

tccaaggcca agccgtggaa tggtagcaat gggatataat acccttctaa gggaaacatt   1320 

tgtatcagta tcatttgatc tgccatggac atgtgtttaa agtggctttc tggcccttct   1380 

ttcaatggct tcttccctaa aacgtggaga ctctaagtta atgtcgttac tatgggccat   1440 

attactaatg cccactgggg tctatgattt ctcaaaattt tcattcggaa tccgaaggat   1500 

acagtcttta aactttagaa ttcccaagaa ggctttatta cacctcagaa attgaaagca   1560 

ccatgacttt gtccattaaa aaattatcca tagttttttt agtgctttta acattccgac   1620 

atacatcatt ctgtgattaa atctccagat ctctgtaaat gatacctaca ttctaaagag   1680 

ttaattctaa ttattccgat atgaccttaa ggaaaagtaa aggaataaat ttttgtcttt   1740 

gttgaagtat ttaatagagt aaggtaaaga agatattaag tccctttcaa aatggaaaat   1800 

taattctaaa ctgagaaaaa tgttcctact acctattgct gatactgtct ttgcataaat   1860 

gaataaaaat aaactttttt tcttca                                        1886 

 
           
             53  
             877  
             DNA  
             Homo sapiens  
             
               misc_feature  
               (1)...(877)  
               n = A,T,C or G  
             
           
            53 

ttyggcacga ggaaatttct aacawtktwt yytttaatag ttagactcat actttatttt     60 

gacaaattta agatagaaaa atatcataat gtgaatatag cagttgctct ttttgtaaca    120 

tggtttggga tgtgcagtga aacttgaaag gacttgcttt acaggtggtc cctcttctgg    180 

ctgggtttca gttaattctg aattatattc cagccattgc atttgcttga aagaatattg    240 

gacacagtaa aaaaaagaac aggtttggca ttcaataata aatattataa agcaatgaac    300 

caaaacaact tttaaaataa ttactgaaag caaacttcag acttcatgat taaagctaag    360 

aactcatatt ttcaaaatag ctttaacagt ttctatcaat atataataca atartaggac    420 

acttattttt aaaaaacaag tgagtagaat cagagtaaat atgatatttc agatgactat    480 

aaacagtaaa catcaattca atatatttat atatcatttc agcaatatac tctktgccca    540 

gctggcgata aaaactgtag ttctatcatc aaaaaatgca tccctgaatg tcatctttga    600 

acttactaag tgctgtcatc atttctacac tccatctttg gagggggtgg cttagggact    660 

cttggtacat gcagatattt agttatggtt ataatgacaa aaagtaaatg tgccaggagt    720 

ctgaagcaga aacgttgcct tactttgtta agtagcttca cattcttttg tctctgtgat    780 

gcctcaggtg aagtcacact aaataattca cacaggtgct aattttgttg ctctgtgtca    840 

gtacctttca gcttctttct tttcttccct tccccac                             877 

 
           
             54  
             1364  
             DNA  
             Homo sapiens  
             
               misc_feature  
               (1)...(1364)  
               n = A,T,C or G  
             
           
            54 

tttttttttt tttttttgat tanattaagg ggctgccagc ccggagaaat acttaagata     60 

tgggtgagaa atccccagac ttttatacaa aagatttcca ctttcaaatc aatgtcagta    120 

gacattgata aaagtatagc agcatcctct actgaggtga tttcatttat tccctgcagc    180 

ccactgataa atatctcact tctcccaaat agtatgtgga ctcccagcta agcagaaaac    240 

tattgtcatt caactgaaga agaggaagat aaaagattgt cttgtttcca tcactgtatt    300 

acttgtgtaa catgattaca taattcttat cctaagagaa agctttcata tttaaaaaaa    360 

agtcttttca gataaaatct gcttgtgtct tgaataatat gaaatacaaa ctttcacttt    420 

attttattgt aaattatraa gagattattg tcttaaataa tatattgagt tagcttcaag    480 

cttcctaaaa tatgaagaga ttgttgtcta aagtcacata ttgacattga gctcagtggc    540 

ctgtttcatc acgtatgtgc tgctacctgt acagcagaca tgccgctcca gtgacattta    600 

taatgacaga agcagggtaa tggtcttgtg tttgacatga tcagttagga tcatagactt    660 

tccctgactc gtagatatta gccttgaatt gggggaaaag argactttga cacattttag    720 

ttattttaat aacagagatt tactcttttg aaaaataaag gtatctaatg tctccctaat    780 

aagtcttctt tccttccaac taaatgacct acacggactt ttattttctt gatcaaagag    840 

gtgtttatta aggacttctg gataactata cttttactct atttttaaag atcacaaagt    900 

aattttaaat gtgaacaggt tcccatacca tgaatgctgg cctcaccttc tctatcatcc    960 

acattttgaa atgcaaagaa agctcccttg taagccatac ttccttcccc actcccatcc   1020 

taggatactt gcccagtgct cattaggcat ttcttattca gatagtccaa atttaggtta   1080 

ttatgcttaa tttgacacat taactaaatg cccagtttta aaatatatcc atcaattcac   1140 

gctgaaatgt gcttctttgt gctatcaaat ggaatagaat acacttattt tttaaacaat   1200 

cccagaatac tgtgtgtaga cttttgttgt gctcaaataa atgtttactt atcttacaaa   1260 

gctcaaatac tggattgtaa ccatgtgatg aagttatcta tgttgtacct aacattgcaa   1320 

attaatcaat aaatctctgt tgtcaaaaaa aaaaaaaaaa aaaa                    1364 

 
           
             55  
             539  
             DNA  
             Homo sapiens  
             
               misc_feature  
               (1)...(539)  
               n = A,T,C or G  
             
           
            55 

ccgggccccc cctcgagggy ttcaatggtc agatggaaca gttgaaaggc gcggtcgaaa     60 

ccctcgccat cacgatcgcg caatctggca ttctggaatt cgtcacaacg atcgtcaccg    120 

ccttgggcaa ctttgtcgat aagctcgccg aggtcagccc ggaaactctg aagtgggtca    180 

cgatcatcgg tggggtggcg gcggtgctag gtccggtggc gatcggcatc ggcgccgtgg    240 

tctctgcgct gggcgccttt ctccctgtca tcgtgcctgt tgcgagcgcc atcggcgctg    300 

tcgtttcggt catcacggcc ggtgccatcc cagccctggc cgggcttgtt gttgccctat    360 

cgcctgtgct cgtgccgctg gcggcggtgg ctgctgcagt cggcgccgtt tatctggtgt    420 

ggaagaactg ggacatgatc gggcccattc tcgccaagct ttataacgga gtgaagacgt    480 

ggctggtcga taagctcggc aaggtgtggg aaactctcaa gagcaagata aaagccgta     539 

 
           
             56  
             510  
             PRT  
             Homo sapiens  
           
            56 

Met Pro Arg Gly Phe Leu Val Lys Arg Ser Lys Lys Ser Thr Pro Val 
                  5                  10                  15 

Ser Tyr Arg Val Arg Gly Gly Glu Asp Gly Asp Arg Ala Leu Leu Leu 
             20                  25                  30 

Ser Pro Ser Cys Gly Gly Ala Arg Ala Glu Pro Pro Ala Pro Ser Pro 
         35                  40                  45 

Val Pro Gly Pro Leu Pro Pro Pro Pro Pro Ala Glu Arg Ala His Ala 
     50                  55                  60 

Ala Leu Ala Ala Ala Leu Ala Cys Ala Pro Gly Pro Gln Pro Pro Pro 
 65                  70                  75                  80 

Gln Gly Pro Arg Ala Ala His Phe Gly Asn Pro Glu Ala Ala His Pro 
                 85                  90                  95 

Ala Pro Leu Tyr Ser Pro Thr Arg Pro Val Ser Arg Glu His Glu Lys 
            100                 105                 110 

His Lys Tyr Phe Glu Arg Ser Phe Asn Leu Gly Ser Pro Val Ser Ala 
        115                 120                 125 

Glu Ser Phe Pro Thr Pro Ala Ala Leu Leu Gly Gly Gly Gly Gly Gly 
    130                 135                 140 

Gly Ala Ser Gly Ala Gly Gly Gly Gly Thr Cys Gly Gly Asp Pro Leu 
145                 150                 155                 160 

Leu Phe Ala Pro Ala Glu Leu Lys Met Gly Thr Ala Phe Ser Ala Gly 
                165                 170                 175 

Ala Glu Ala Ala Arg Gly Pro Gly Pro Gly Pro Pro Leu Pro Pro Ala 
            180                 185                 190 

Ala Ala Leu Arg Pro Pro Gly Lys Arg Pro Pro Pro Pro Thr Ala Ala 
        195                 200                 205 

Glu Pro Pro Ala Lys Ala Val Lys Ala Pro Gly Ala Lys Lys Pro Lys 
    210                 215                 220 

Ala Ile Arg Lys Leu His Phe Glu Asp Glu Val Thr Thr Ser Pro Val 
225                 230                 235                 240 

Leu Gly Leu Lys Ile Lys Glu Gly Pro Val Glu Ala Pro Arg Gly Arg 
                245                 250                 255 

Ala Gly Gly Ala Ala Arg Pro Leu Gly Glu Phe Ile Cys Gln Leu Cys 
            260                 265                 270 

Lys Glu Glu Tyr Ala Asp Pro Phe Ala Leu Ala Gln His Lys Cys Ser 
        275                 280                 285 

Arg Ile Val Arg Val Glu Tyr Arg Cys Pro Glu Cys Ala Lys Val Phe 
    290                 295                 300 

Ser Cys Pro Ala Asn Leu Ala Ser His Arg Arg Trp His Lys Pro Arg 
305                 310                 315                 320 

Pro Ala Pro Ala Ala Ala Arg Ala Pro Glu Pro Glu Ala Ala Ala Arg 
                325                 330                 335 

Ala Glu Ala Arg Glu Ala Pro Gly Gly Gly Ser Asp Arg Asp Thr Pro 
            340                 345                 350 

Ser Pro Gly Gly Val Ser Glu Ser Gly Ser Glu Asp Gly Leu Tyr Glu 
        355                 360                 365 

Cys His His Cys Ala Lys Lys Phe Arg Arg Gln Ala Tyr Leu Arg Lys 
    370                 375                 380 

His Leu Leu Ala His His Gln Ala Leu Gln Ala Lys Gly Ala Pro Leu 
385                 390                 395                 400 

Ala Pro Pro Ala Glu Asp Leu Leu Ala Leu Tyr Pro Gly Pro Asp Glu 
                405                 410                 415 

Lys Ala Pro Gln Glu Ala Ala Gly Asp Gly Glu Gly Ala Gly Val Leu 
            420                 425                 430 

Gly Leu Ser Ala Ser Ala Glu Cys His Leu Cys Pro Val Cys Gly Glu 
        435                 440                 445 

Ser Phe Ala Ser Lys Gly Ala Gln Glu Arg His Leu Arg Leu Leu His 
    450                 455                 460 

Ala Ala Gln Val Phe Pro Cys Lys Tyr Cys Pro Ala Thr Phe Tyr Ser 
465                 470                 475                 480 

Ser Pro Gly Leu Thr Arg His Ile Asn Lys Cys His Pro Ser Glu Asn 
                485                 490                 495 

Arg Gln Val Ile Leu Leu Gln Val Pro Val Arg Pro Ala Cys 
            500                 505                 510 

 
           
             57  
             1047  
             PRT  
             Homo sapiens  
           
            57 

Met Asp Ala Glu Ala Glu Asp Lys Thr Leu Arg Thr Arg Ser Lys Gly 
                  5                  10                  15 

Thr Glu Val Pro Met Asp Ser Leu Ile Gln Glu Leu Ser Val Ala Tyr 
             20                  25                  30 

Asp Cys Ser Met Ala Lys Lys Arg Thr Ala Glu Asp Gln Ala Leu Gly 
         35                  40                  45 

Val Pro Val Asn Lys Arg Lys Ser Leu Leu Met Lys Pro Arg His Tyr 
     50                  55                  60 

Ser Pro Lys Ala Asp Cys Gln Glu Asp Arg Ser Asp Arg Thr Glu Asp 
 65                  70                  75                  80 

Asp Gly Pro Leu Glu Thr His Gly His Ser Thr Ala Glu Glu Ile Met 
                 85                  90                  95 

Ile Lys Pro Met Asp Glu Ser Leu Leu Ser Thr Ala Gln Glu Asn Ser 
            100                 105                 110 

Ser Arg Lys Glu Asp Arg Tyr Ser Cys Tyr Gln Glu Leu Met Val Lys 
        115                 120                 125 

Ser Leu Met His Leu Gly Lys Phe Glu Lys Asn Val Ser Val Gln Thr 
    130                 135                 140 

Val Ser Glu Asn Leu Asn Asp Ser Gly Ile Gln Ser Leu Lys Ala Glu 
145                 150                 155                 160 

Ser Asp Glu Ala Asp Glu Cys Phe Leu Ile His Ser Asp Asp Gly Arg 
                165                 170                 175 

Asp Lys Ile Asp Asp Ser Gln Pro Pro Phe Cys Ser Ser Asp Asp Asn 
            180                 185                 190 

Glu Ser Asn Ser Glu Ser Ala Glu Asn Gly Trp Asp Ser Gly Ser Asn 
        195                 200                 205 

Phe Ser Glu Glu Thr Lys Pro Pro Arg Val Pro Lys Tyr Val Leu Thr 
    210                 215                 220 

Asp His Lys Lys Asp Leu Leu Glu Val Pro Glu Ile Lys Thr Glu Gly 
225                 230                 235                 240 

Asp Lys Phe Ile Pro Cys Glu Asn Arg Cys Asp Ser Glu Thr Glu Arg 
                245                 250                 255 

Lys Asp Pro Gln Asn Ala Leu Ala Glu Pro Leu Asp Gly Asn Ala Gln 
            260                 265                 270 

Pro Ser Phe Pro Asp Val Glu Glu Glu Asp Ser Glu Ser Leu Ala Val 
        275                 280                 285 

Met Thr Glu Glu Gly Ser Asp Leu Glu Lys Ala Lys Gly Asn Leu Ser 
    290                 295                 300 

Leu Leu Glu Gln Ala Ile Ala Leu Gln Ala Glu Arg Gly Cys Val Phe 
305                 310                 315                 320 

His Asn Thr Tyr Lys Glu Leu Asp Arg Phe Leu Leu Glu His Leu Ala 
                325                 330                 335 

Gly Glu Arg Arg Gln Thr Lys Val Ile Asp Met Gly Gly Arg Gln Ile 
            340                 345                 350 

Phe Asn Asn Lys His Ser Pro Arg Pro Glu Lys Arg Glu Thr Lys Cys 
        355                 360                 365 

Pro Ile Pro Gly Cys Asp Gly Thr Gly His Val Thr Gly Leu Tyr Pro 
    370                 375                 380 

His His Arg Ser Leu Ser Gly Cys Pro His Lys Val Arg Val Pro Leu 
385                 390                 395                 400 

Glu Ile Leu Ala Met His Glu Asn Val Leu Lys Cys Pro Thr Pro Gly 
                405                 410                 415 

Cys Thr Gly Arg Gly His Val Asn Ser Asn Arg Asn Thr His Arg Ser 
            420                 425                 430 

Leu Ser Gly Cys Pro Ile Ala Ala Ala Glu Lys Leu Ala Met Ser Gln 
        435                 440                 445 

Asp Lys Asn Gln Leu Asp Ser Pro Gln Thr Gly Gln Cys Pro Asp Gln 
    450                 455                 460 

Ala His Arg Thr Ser Leu Val Lys Gln Ile Glu Phe Asn Phe Pro Ser 
465                 470                 475                 480 

Gln Ala Ile Thr Ser Pro Arg Ala Thr Val Ser Lys Glu Gln Glu Lys 
                485                 490                 495 

Phe Gly Lys Val Pro Phe Asp Tyr Ala Ser Phe Asp Ala Gln Val Phe 
            500                 505                 510 

Gly Lys Arg Pro Leu Ile Gln Thr Val Gln Gly Arg Lys Thr Pro Pro 
        515                 520                 525 

Phe Pro Glu Ser Lys His Phe Pro Asn Pro Val Lys Phe Pro Asn Arg 
    530                 535                 540 

Leu Pro Ser Ala Gly Ala His Thr Gln Ser Pro Gly Arg Ala Ser Ser 
545                 550                 555                 560 

Tyr Ser Tyr Gly Gln Cys Ser Glu Asp Thr His Ile Ala Ala Ala Ala 
                565                 570                 575 

Ala Ile Leu Asn Leu Ser Thr Arg Cys Arg Glu Ala Thr Asp Ile Leu 
            580                 585                 590 

Ser Asn Lys Pro Gln Ser Leu His Ala Lys Gly Ala Glu Ile Glu Val 
        595                 600                 605 

Asp Glu Asn Gly Thr Leu Asp Leu Ser Met Lys Lys Asn Arg Ile Leu 
    610                 615                 620 

Asp Lys Ser Ala Pro Leu Thr Ser Ser Asn Thr Ser Ile Pro Thr Pro 
625                 630                 635                 640 

Ser Ser Ser Pro Phe Lys Thr Ser Ser Ile Leu Val Asn Ala Ala Phe 
                645                 650                 655 

Tyr Gln Ala Leu Cys Asp Gln Glu Gly Trp Asp Thr Pro Ile Asn Tyr 
            660                 665                 670 

Ser Lys Thr His Gly Lys Thr Glu Glu Glu Lys Glu Lys Asp Pro Val 
        675                 680                 685 

Ser Ser Leu Glu Asn Leu Glu Glu Lys Lys Phe Pro Gly Glu Ala Ser 
    690                 695                 700 

Ile Pro Ser Pro Lys Pro Lys Leu His Ala Arg Asp Leu Lys Lys Glu 
705                 710                 715                 720 

Leu Ile Thr Cys Pro Thr Pro Gly Cys Asp Gly Ser Gly His Val Thr 
                725                 730                 735 

Gly Asn Tyr Ala Ser His Arg Ser Val Ser Gly Cys Pro Leu Ala Asp 
            740                 745                 750 

Lys Thr Leu Lys Ser Leu Met Ala Ala Asn Ser Gln Glu Leu Lys Cys 
        755                 760                 765 

Pro Thr Pro Gly Cys Asp Gly Ser Gly His Val Thr Gly Asn Tyr Ala 
    770                 775                 780 

Ser His Arg Ser Leu Ser Gly Cys Pro Arg Ala Arg Lys Gly Gly Val 
785                 790                 795                 800 

Lys Met Thr Pro Thr Lys Glu Glu Lys Glu Asp Pro Glu Leu Lys Cys 
                805                 810                 815 

Pro Val Ile Gly Cys Asp Gly Gln Gly His Ile Ser Gly Lys Tyr Thr 
            820                 825                 830 

Ser His Arg Thr Ala Ser Gly Cys Pro Leu Ala Ala Lys Arg Gln Lys 
        835                 840                 845 

Glu Asn Pro Leu Asn Gly Ala Ser Leu Ser Trp Lys Leu Asn Lys Gln 
    850                 855                 860 

Glu Leu Pro His Cys Pro Leu Pro Gly Cys Asn Gly Leu Gly His Val 
865                 870                 875                 880 

Asn Asn Val Phe Val Thr His Arg Ser Leu Ser Gly Cys Pro Leu Asn 
                885                 890                 895 

Ala Gln Val Ile Lys Lys Gly Lys Val Ser Glu Glu Leu Met Thr Ile 
            900                 905                 910 

Lys Leu Lys Ala Thr Gly Gly Ile Glu Ser Asp Glu Glu Ile Arg His 
        915                 920                 925 

Leu Asp Glu Glu Ile Lys Glu Leu Asn Glu Ser Asn Leu Lys Ile Glu 
    930                 935                 940 

Ala Asp Met Met Lys Leu Gln Thr Gln Ile Thr Ser Met Glu Ser Asn 
945                 950                 955                 960 

Leu Lys Thr Ile Glu Glu Glu Asn Lys Leu Ile Glu Gln Asn Asn Glu 
                965                 970                 975 

Ser Leu Leu Lys Glu Leu Ala Gly Leu Ser Gln Ala Leu Ile Ser Ser 
            980                 985                 990 

Leu Ala Asp Ile Gln Leu Pro Gln Met Gly Pro Ile Ser Glu Gln Asn 
        995                 1000                1005 

Phe Glu Ala Tyr Val Asn Thr Leu Thr Asp Met Tyr Ser Asn Leu Glu 
    1010                1015                1020 

Arg Asp Tyr Ser Pro Glu Cys Lys Ala Leu Leu Glu Ser Ile Lys Gln 
1025                1030                1035                1040 

Ala Val Lys Gly Ile His Val 
                1045 

 
           
             58  
             2165  
             DNA  
             Homo sapiens  
           
            58 

cgccaccgct gggtgcggcg aggccggcgc gatgcggcag ctgtgccggg gccgcgtgct     60 

gggcatctcg gtggccatcg cgcacggggt cttctcgggc tccctcaaca tcttgctcaa    120 

gttcctcatc agccgctacc agttctcctt cctgaccctg gtgcagtgcc tgaccagctc    180 

caccgcggcg ctgagcctgg agctgctgcg gcgcctcggg ctcatcgccg tgcccccctt    240 

cggtctgagc ctggcgcgct ccttcgcggg ggtcgcggtg ctctccacgc tgcagtccag    300 

cctcacgctc tggtccctgc gcggcctcag cctgcccatg tacgtggtct tcaagcgctg    360 

cctgcccctg gtcaccatgc tcatcggcgt cctggtgctc aagaacggcg cgccctcgcc    420 

aggggtgctg gcggcggtgc tcatcaccac ctgcggcgcc gccctggcag gagccggcga    480 

cctgacgggc gaccccatcg ggtacgtcac gggagtgctg gcggtgctgg tgcacgctgc    540 

ctacctggtg ctcatccaga aggccagcgc agacaccgag cacgggccgc tcaccgcgca    600 

gtacgtcatc gccgtctctg ccaccccgct gctggtcatc tgctccttcg ccagcaccga    660 

ctccatccac gcctggacct tcccgggctg gaaggacccg gccatggtct gcatcttcgt    720 

ggcctgcatc ctgatcggct gcgccatgaa cttcaccacg ctgcactgca cctacatcaa    780 

ttcggccgtg accacctctc tgttcattgc cggcgtggtg gtgaacaccc tgggctctat    840 

catttactgt gtggccaagt tcatggagac cagaaagcaa agcaactacg aggacctgga    900 

ggcccagcct cggggagagg aggcgcagct aagtggagac cagctgccgt tcgtgatgga    960 

ggagctgccc ggggagggag gaaatggccg gtcagaaggt ggggaggcag caggtggccc   1020 

cgctcaggag agcaggcaag aggtcagggg cagcccccga ggagtcccgc tggtggctgg   1080 

gagctctgaa gaagggagca ggaggtcgtt aaaagatgct tacctcgagg tatggaggtt   1140 

ggttagggga accaggtata tgaagaagga ttatttgata gaaaacgagg agttacccag   1200 

tccttgagaa ggaggtgcat gtacgtacct atgtgcatac acttatttta tatgttagaa   1260 

atgacgtgtt ttaatgagag gcctccccgt tttattcttt gaggagtggg gaagggaaga   1320 

aaagaaagaa gctgaaaggt actgacacag agcaacaaaa ttagcacctg tgtgaattat   1380 

ttagtgtgac ttcacctgag gcatcacaga gacaaaagaa tgtgaagcta cttaacaaag   1440 

taaggcaacg tttctgcttc agactcctgg cacatttact ttttgtcatt ataaccataa   1500 

ctaaatatct gcatgtacca agagtcccta agccaccccc tccaaagatg gagtgtagaa   1560 

atgatgacag cacttagtaa gttcaaagat gacattcagg gatgcatttt ttgatgatag   1620 

aactacagtt tttatcgcca gctgggcaaa gagtatattg ctgaaatgat atataaatat   1680 

attgaattga tgtttactgt ttatagtcat ctgaaatatc atatttactc tgattctact   1740 

cacttgtttt ttaaaaataa gtgtcctact attgtattat atattgatag aaactgttaa   1800 

agctattttg aaaatatgag ttcttagctt taatcatgaa gtctgaagtt tgctttcagt   1860 

aattatttta aaagttgttt tggttcattg ctttataata tttattattg aatgccaaac   1920 

ctgttctttt ttttactgtg tccaatattc tttcaagcaa atgcaatggc tggaatataa   1980 

ttcagaatta actgaaaccc agccagaaga gggaccacct gtaaagcaag tcctttcaag   2040 

tttcactgca catcccaaac catgttacaa aaagagcaac tgctatattc acattatgat   2100 

atttttctat cttaaatttg tcaaaataaa gtatgagtct aactattaaa aaaaaaaaaa   2160 

aaaaa                                                               2165 

 
           
             59  
             1176  
             DNA  
             Homo sapiens  
           
            59 

atgcggcagc tgtgccgggg ccgcgtgctg ggcatctcgg tggccatcgc gcacggggtc     60 

ttctcgggct ccctcaacat cttgctcaag ttcctcatca gccgctacca gttctccttc    120 

ctgaccctgg tgcagtgcct gaccagctcc accgcggcgc tgagcctgga gctgctgcgg    180 

cgcctcgggc tcatcgccgt gccccccttc ggtctgagcc tggcgcgctc cttcgcgggg    240 

gtcgcggtgc tctccacgct gcagtccagc ctcacgctct ggtccctgcg cggcctcagc    300 

ctgcccatgt acgtggtctt caagcgctgc ctgcccctgg tcaccatgct catcggcgtc    360 

ctggtgctca agaacggcgc gccctcgcca ggggtgctgg cggcggtgct catcaccacc    420 

tgcggcgccg ccctggcagg agccggcgac ctgacgggcg accccatcgg gtacgtcacg    480 

ggagtgctgg cggtgctggt gcacgctgcc tacctggtgc tcatccagaa ggccagcgca    540 

gacaccgagc acgggccgct caccgcgcag tacgtcatcg ccgtctctgc caccccgctg    600 

ctggtcatct gctccttcgc cagcaccgac tccatccacg cctggacctt cccgggctgg    660 

aaggacccgg ccatggtctg catcttcgtg gcctgcatcc tgatcggctg cgccatgaac    720 

ttcaccacgc tgcactgcac ctacatcaat tcggccgtga ccacctctct gttcattgcc    780 

ggcgtggtgg tgaacaccct gggctctatc atttactgtg tggccaagtt catggagacc    840 

agaaagcaaa gcaactacga ggacctggag gcccagcctc ggggagagga ggcgcagcta    900 

agtggagacc agctgccgtt cgtgatggag gagctgcccg gggagggagg aaatggccgg    960 

tcagaaggtg gggaggcagc aggtggcccc gctcaggaga gcaggcaaga ggtcaggggc   1020 

agcccccgag gagtcccgct ggtggctggg agctctgaag aagggagcag gaggtcgtta   1080 

aaagatgctt acctcgaggt atggaggttg gttaggggaa ccaggtatat gaagaaggat   1140 

tatttgatag aaaacgagga gttacccagt ccttga                             1176 

 
           
             60  
             1089  
             DNA  
             Homo sapiens  
           
            60 

cgccaccgct gggtgcggcg aggccggcgc gatgcggcag ctgtgccggg gccgcgtgct     60 

gggcatctcg gtggccatcg cgcacggggt cttctcgggc tccctcaaca tcttgctcaa    120 

gttcctcatc agccgctacc agttctcctt cctgaccctg gtgcagtgcc tgaccagctc    180 

caccgcggcg ctgagcctgg agctgctgcg gcgcctcggg ctcatcgccg tgcccccctt    240 

cggtctgagc ctggcgcgct ccttcgcggg ggtcgcggtg ctctccacgc tgcagtccag    300 

cctcacgctc tggtccctgc gcggcctcag cctgcccatg tacgtggtct tcaagcgctg    360 

cctgcccctg gtcaccatgc tcatcggcgt cctggtgctc aagaacggcg cgccctcgcc    420 

aggggtgctg gcggcggtgc tcatcaccac ctgcggcgcc gccctggcag gagccggcga    480 

cctgacgggc gaccccatcg ggtacgtcac gggagtgctg gcggtgctgg tgcacgctgc    540 

ctacctggtg ctcatccaga aggccagcgc agacaccgag cacgggccgc tcaccgcgca    600 

gtacgtcatc gccgtctctg ccaccccgct gctggtcatc tgctccttcg ccagcaccga    660 

ctccatccac gcctggacct tcccgggctg gaaggacccg gccatggtct gcatcttcgt    720 

ggcctgcatc ctgatcggct gcgccatgaa cttcaccacg ctgcactgca cctacatcaa    780 

ttcggccgtg accacctctc tgttcattgc cggcgtggtg gtgaacaccc tgggctctat    840 

catttactgt gtggccaagt tcatggagac cagaaagcaa agcaactacg aggacctgga    900 

ggcccagcct cggggagagg aggcgcagct aagtggagac cagctgccgt tcgtgatgga    960 

ggagctgccc ggggagggag gaaatggccg gtcagaaggt ggggaggcag caggtggccc   1020 

cgctcaggag agcaggcaag aggtcagggg cagcccccga ggagtcccgc tggtggctgg   1080 

gagctctga                                                           1089 

 
           
             61  
             362  
             PRT  
             Homo sapiens  
           
            61 

Arg His Arg Trp Val Arg Arg Gly Arg Arg Asp Ala Ala Ala Val Pro 
                  5                  10                  15 

Gly Pro Arg Ala Gly His Leu Gly Gly His Arg Ala Arg Gly Leu Leu 
             20                  25                  30 

Gly Leu Pro Gln His Leu Ala Gln Val Pro His Gln Pro Leu Pro Val 
         35                  40                  45 

Leu Leu Pro Asp Pro Gly Ala Val Pro Asp Gln Leu His Arg Gly Ala 
     50                  55                  60 

Glu Pro Gly Ala Ala Ala Ala Pro Arg Ala His Arg Arg Ala Pro Leu 
 65                  70                  75                  80 

Arg Ser Glu Pro Gly Ala Leu Leu Arg Gly Gly Arg Gly Ala Leu His 
                 85                  90                  95 

Ala Ala Val Gln Pro His Ala Leu Val Pro Ala Arg Pro Gln Pro Ala 
            100                 105                 110 

His Val Arg Gly Leu Gln Ala Leu Pro Ala Pro Gly His His Ala His 
        115                 120                 125 

Arg Arg Pro Gly Ala Gln Glu Arg Arg Ala Leu Ala Arg Gly Ala Gly 
    130                 135                 140 

Gly Gly Ala His His His Leu Arg Arg Arg Pro Gly Arg Ser Arg Arg 
145                 150                 155                 160 

Pro Asp Gly Arg Pro His Arg Val Arg His Gly Ser Ala Gly Gly Ala 
                165                 170                 175 

Gly Ala Arg Cys Leu Pro Gly Ala His Pro Glu Gly Gln Arg Arg His 
            180                 185                 190 

Arg Ala Arg Ala Ala His Arg Ala Val Arg His Arg Arg Leu Cys His 
        195                 200                 205 

Pro Ala Ala Gly His Leu Leu Leu Arg Gln His Arg Leu His Pro Arg 
    210                 215                 220 

Leu Asp Leu Pro Gly Leu Glu Gly Pro Gly His Gly Leu His Leu Arg 
225                 230                 235                 240 

Gly Leu His Pro Asp Arg Leu Arg His Glu Leu His His Ala Ala Leu 
                245                 250                 255 

His Leu His Gln Phe Gly Arg Asp His Leu Ser Val His Cys Arg Arg 
            260                 265                 270 

Gly Gly Glu His Pro Gly Leu Tyr His Leu Leu Cys Gly Gln Val His 
        275                 280                 285 

Gly Asp Gln Lys Ala Lys Gln Leu Arg Gly Pro Gly Gly Pro Ala Ser 
    290                 295                 300 

Gly Arg Gly Gly Ala Ala Lys Trp Arg Pro Ala Ala Val Arg Asp Gly 
305                 310                 315                 320 

Gly Ala Ala Arg Gly Gly Arg Lys Trp Pro Val Arg Arg Trp Gly Gly 
                325                 330                 335 

Ser Arg Trp Pro Arg Ser Gly Glu Gln Ala Arg Gly Gln Gly Gln Pro 
            340                 345                 350 

Pro Arg Ser Pro Ala Gly Gly Trp Glu Leu 
        355                 360 

 
           
             62  
             391  
             PRT  
             Homo sapiens  
           
            62 

Met Arg Gln Leu Cys Arg Gly Arg Val Leu Gly Ile Ser Val Ala Ile 
                  5                  10                  15 

Ala His Gly Val Phe Ser Gly Ser Leu Asn Ile Leu Leu Lys Phe Leu 
             20                  25                  30 

Ile Ser Arg Tyr Gln Phe Ser Phe Leu Thr Leu Val Gln Cys Leu Thr 
         35                  40                  45 

Ser Ser Thr Ala Ala Leu Ser Leu Glu Leu Leu Arg Arg Leu Gly Leu 
     50                  55                  60 

Ile Ala Val Pro Pro Phe Gly Leu Ser Leu Ala Arg Ser Phe Ala Gly 
 65                  70                  75                  80 

Val Ala Val Leu Ser Thr Leu Gln Ser Ser Leu Thr Leu Trp Ser Leu 
                 85                  90                  95 

Arg Gly Leu Ser Leu Pro Met Tyr Val Val Phe Lys Arg Cys Leu Pro 
            100                 105                 110 

Leu Val Thr Met Leu Ile Gly Val Leu Val Leu Lys Asn Gly Ala Pro 
        115                 120                 125 

Ser Pro Gly Val Leu Ala Ala Val Leu Ile Thr Thr Cys Gly Ala Ala 
    130                 135                 140 

Leu Ala Gly Ala Gly Asp Leu Thr Gly Asp Pro Ile Gly Tyr Val Thr 
145                 150                 155                 160 

Gly Val Leu Ala Val Leu Val His Ala Ala Tyr Leu Val Leu Ile Gln 
                165                 170                 175 

Lys Ala Ser Ala Asp Thr Glu His Gly Pro Leu Thr Ala Gln Tyr Val 
            180                 185                 190 

Ile Ala Val Ser Ala Thr Pro Leu Leu Val Ile Cys Ser Phe Ala Ser 
        195                 200                 205 

Thr Asp Ser Ile His Ala Trp Thr Phe Pro Gly Trp Lys Asp Pro Ala 
    210                 215                 220 

Met Val Cys Ile Phe Val Ala Cys Ile Leu Ile Gly Cys Ala Met Asn 
225                 230                 235                 240 

Phe Thr Thr Leu His Cys Thr Tyr Ile Asn Ser Ala Val Thr Thr Ser 
                245                 250                 255 

Leu Phe Ile Ala Gly Val Val Val Asn Thr Leu Gly Ser Ile Ile Tyr 
            260                 265                 270 

Cys Val Ala Lys Phe Met Glu Thr Arg Lys Gln Ser Asn Tyr Glu Asp 
        275                 280                 285 

Leu Glu Ala Gln Pro Arg Gly Glu Glu Ala Gln Leu Ser Gly Asp Gln 
    290                 295                 300 

Leu Pro Phe Val Met Glu Glu Leu Pro Gly Glu Gly Gly Asn Gly Arg 
305                 310                 315                 320 

Ser Glu Gly Gly Glu Ala Ala Gly Gly Pro Ala Gln Glu Ser Arg Gln 
                325                 330                 335 

Glu Val Arg Gly Ser Pro Arg Gly Val Pro Leu Val Ala Gly Ser Ser 
            340                 345                 350 

Glu Glu Gly Ser Arg Arg Ser Leu Lys Asp Ala Tyr Leu Glu Val Trp 
        355                 360                 365 

Arg Leu Val Arg Gly Thr Arg Tyr Met Lys Lys Asp Tyr Leu Ile Glu 
    370                 375                 380 

Asn Glu Glu Leu Pro Ser Pro 
385                 390 

 
           
             63  
             442  
             DNA  
             Homo sapiens  
             
               misc_feature  
               220,391,428  
               n = A,T,C or G  
             
           
            63 

atagtaagca ctgatgtgtt tattcgatga aataggggtg ggggtgtagc agccctagtc     60 

ccacattgca tgggctggtg actgagttaa cagcaaagtg ggatgcaaaa ggttcctgat    120 

tggagacccc cggattcggg ttctggattt gctggccact tactctatga cttggggcat    180 

gtcactgtca tggcctcagt ttccccttct gcacagtgtn ttattggata gttccagctc    240 

tgacatgcta ggattatgtg atactgtcaa tcaagactag ggttggccta agcacatggt    300 

ctgaaaacac ctcgggctca tggacatatt ttctccgcat ggggagtggg cagctgctga    360 

gtggcaaggc tgccctccaa agctgtccat nccacgcccg gggtgctgtg ggtctccttt    420 

ccctcgtngc cgaattcttg gg                                             442 

 
           
             64  
             456  
             DNA  
             Homo sapiens  
           
            64 

cttcaaccat aaaaacaaag ggctctgatt gctttagggg ataagtgatt taatatccac     60 

aaacgtcccc actcccaaaa gtaactatat tctggatttc aacttttctt ctaattgtga    120 

atccttctgt tttttcttct taaggaggaa agttaaagga cactacaggt catcaaaaac    180 

aagttggcca aggactcatt acttgtctta tatttttact gccactaaac tgcctgtatt    240 

tctgtatgtc cttctatcca aacagacgtt cactgccact tgtaaagtga aggatgtaaa    300 

cgaggatata taactgtttc agtgaacaga ttttgtgaag tgccttctgt tttagcactt    360 

taagtttatc acattttgtt gacttctgac attccacttt cctaggttat aggaaagatc    420 

tgtttatgta gtttgttttt aaaatgtgcc aatgcc                              456 

 
           
             65  
             654  
             DNA  
             Homo sapiens  
           
            65 

aataaattcc agccttctct ttcttgctgc ttcctcagat attttcctcc tttcttctcc     60 

agtattcact ctcttctctg gagtttgatg ggcctgttta tgtttttgca gtggtttctt    120 

ttcgtgtaat tttttatctc catatttctt atatgctaaa ggtattccat atttagcggc    180 

aggctttgta attttctgag caggcataac agaaatcgag ttttgtcctg aagctggtct    240 

tttagctggt ataggctgtg atccaaactt cgaaaatgtt tttagacaaa attcttctgc    300 

aataagctga ggagagagaa acttttcaat gcgtttggct ataaaacctt tctccaatat    360 

ggagttgact gatggtctat ccctaggatt tcttttaaat aactgagaca ccaaactgcg    420 

gagatcatag gaataatgca aagacacagg tggaaaagat ccagatatta tcttcagtac    480 

caggtttttc atactgccag cttcaaaagc atgtttaagt gtacacagct cataaaggac    540 

acaccccaga gcccaaatgt ccttttatta ttgtaagttt gttttcacag atttcaggtg    600 

acaagtagat tggggcccct atcaagttcg gccccctctc cagtctttta gaac          654 

 
           
             66  
             592  
             DNA  
             Homo sapiens  
           
            66 

tttttttttt tttttttatt gggaataaat ttatcaaaaa acatgtcatc caattcccac     60 

aaatgagaca ttttaaatac agaatacact ctgttcatga atataaaatc cccaggtgaa    120 

agtcccttaa aacactatta tggttatgtt tcctagaata attttataac tttttcagag    180 

aattccttta aacttgttaa aataccttgt tgctagtgct cagaacatct aggttcagtc    240 

tttattttta agacagtatc tatcctaggc aaatgagagc ttgtttttat gtatttaaga    300 

gtttcctctt gtcatttcaa tgtcaaattg atttgactca atttcatgat ttcatctcgc    360 

tcaaggccat caaccggtca gagccagagc ccttcaaagg ctgtatgtga gtatatgagg    420 

gaaaactttc cacataattt tacatcattt ctatctcata gcagttttag ttttctcata    480 

gctatctcat agcagtttta gttttctcaa attctatgct gtttttgtac tactgcagct    540 

gaccaatcca aagccagttt acactcagca tgtgttattc tactttaaaa ta            592 

 
           
             67  
             469  
             DNA  
             Homo sapiens  
             
               misc_feature  
               245,298,314,339,424,440,465  
               n = A,T,C or G  
             
           
            67 

gatgccaaaa atgctttccc aagtggctaa cattctgtat tcccaccagc aatatatgag     60 

agattaagtt gcttttcaaa cccatttatg ctcagtattg tcaggttttg ttttgttctg    120 

ggttctttat ttgttggttt tcttttttat ttcagccatg ctaataggtg tgattgtggt    180 

tttaatttgc aattccctaa cttcataaat tagggaacac agaacacaca tatgacacag    240 

aaaantgcat ttgacctgat tttacttcct actattaaga aacagataaa attcatantg    300 

tccctggaac accntttttt tgttgcttta tttgtcatna catttaatct tttgttaagt    360 

ggaaatggtc tcttcagata atttttttcc attttaaatc aggttggttg acctatacat    420 

tgtngttttg agagttccan aaggtatccc gtattccaaa tcctncatt                469 

 
           
             68  
             510  
             DNA  
             Homo sapiens  
             
               misc_feature  
               424,462  
               n = A,T,C or G  
             
           
            68 

tttttcctga gaatttaatt ttatttgctg tagattcaaa atgaggaagt ggtaaatgca     60 

ttatttactc aaagcataaa gtcagcctta ggtaggagat gtaacaactc ctcaacttta    120 

cactatccag ttaaagccaa tttttaaaac cttttttttc cttatgatga cccttgagtc    180 

atagaaaact tttcatttta gaaaatgtta agcatgaaca caaaaagact acgataacag    240 

tgttataaac actcgtgtac ccaaggccca gctttaacat tcatcactta gcatgtttaa    300 

ggtagtgctt aggttgaaat ttatattgtg tgtatcagaa taaagagcag ttcttgcaga    360 

tagctagaat tacttcattt ttataggagt ttagagcata aactaacaag ggaatctagg    420 

cccnttatag taaatatcct aaaagcattt taattttaca gnattggaca gcggtatgcc    480 

atggacctat tcccatttgg tcaggggcaa                                     510 

 
           
             69  
             483  
             DNA  
             Homo sapiens  
           
            69 

tgcatcagtt aatgtaatca gcccacagga tggggattga atggaagtat gcccagtacc     60 

tttaagatat gaagctggtc tgaagtacac cttgaacaat atatgtacag ttcatcacac    120 

actgtattta tttgctggag tgtaaattct cggagaacag aatttaagac ttggggcaaa    180 

cagagtctct tttctcctcc aacttgaaaa caagaaatag attccccttc caacacagtc    240 

tgagtgagtt ctgtggagct atctgaaggg atgagcaatg ggccaggaag aacctgaggt    300 

gatggaagag gcagaaatac agtaggcgac atgctttctt gggaatgccg agcagaaaat    360 

gctgctggtc caccagcgag ctctgactac tttaatggaa ttgtgccatg tgtgtttcaa    420 

actgggatta aatggcaatt ttagggaacg agtacaggtc gcctacatgg ctccatcagt    480 

ttc                                                                  483 

 
           
             70  
             481  
             DNA  
             Homo sapiens  
           
            70 

gtactggaca gacgtgagcg aggaggccat caagcagacc tacctgaacc agacgggggc     60 

cgccgtgcag aacgtggtca tctccggcct ggtctctccc gacggcctcg cctgcgactg    120 

ggtgggcaag aagctgtact ggacggactc agagaccaac cgcatcgagg tggccaacct    180 

caatggcaca tcccggaagg tgctcttctg gcaggacctt gaccagccga gggccatcgc    240 

cttggacccc gctcacgggt acatgtactg gacagactgg ggtgagacgc cccggattga    300 

gcgggcaggg atggatggca gcacccggaa gatcattgtg gactcggaca tttactggcc    360 

caatggactg accatcgacc tggaggagca gaagctctac tgggctgacg ccaagctcag    420 

cttcatccac cgtgccaacc tggacggctc gttccggcag aaggtggtgg agggcagcct    480 

g                                                                    481 

 
           
             71  
             341  
             DNA  
             Homo sapiens  
           
            71 

cggccgcggc gaggctggag aagtagtgct ggccgggcga gtcgctccag caggccgggg     60 

acgcgggcgc ggcagggggc gtggggcccg gctctggtgg ggggtcctgg gcccgcacat    120 

agctgcgaag ggtgatgtcg gccgagcccc ctgactccag tgggatgggg tgtgtgtgga    180 

agtggcggag catgtcaagc acagactgga accacagatg ctgtacgtga cactggccgt    240 

ggccgttcag ggacaggcgc atgtgcttgg ccttgccctg gaagttgaag gtcagcacgt    300 

actccccagg ccgagtctca ctttggcggc ccctcgtgcc g                        341 

 
           
             72  
             283  
             DNA  
             Homo sapiens  
           
            72 

tttttagatc catccattta ttccttcagc caacattttc tgggattcct tgtgtgctag     60 

gcctcgtgcc accatctgga gatgcagaga ggcgggagac ccatgtggcc tttgaggggc    120 

tttcaggctc gtgggggttc aggcacagac accaccaatc tgaaccaggg gactgcagga    180 

tgctgggtta ggggagagag ggataggctg gctggcctag ggggtcctca ggaagtcttt    240 

gggggtaagg agagaactcc tgaaaggtaa ggagaagccg agg                      283 

 
           
             73  
             485  
             DNA  
             Homo sapiens  
           
            73 

ttttttttat ttttaggata ttttatttta atgcaaatga aatttctatc tatgtgaaac     60 

tggtaaaggg gagatatagg aactcctatt tttctctctg tcttcctctc tgtttcttct    120 

ttttttattt atttttggat tatagatgct cctctcagtt gcaagttgca atgctccaca    180 

tctctcagcc agcacctggc tctgttccag ggcttttagt gagtgctctc tgtcaaggca    240 

tgaataatac agcccctagg ctgttggcag actccaaatg aggcgtgcat acatcaggaa    300 

gcaagccctt gactttagct ccagaacagc ctccttctgt gtcttgcata tttgccactg    360 

acatgaccac tgccgtcaca gccaggggtg ggacagctga acagctcttg tatggctggt    420 

tccacgggaa ctcgaacccc tttggaccgc gtgcgatgcc gcttctcctc ggtgtgcaac    480 

tccat                                                                485 

 
           
             74  
             338  
             DNA  
             Homo sapiens  
           
            74 

ttttttgatt atttcagaga tttattgcaa gttaattgtc tgtgaagctg gatattcctt     60 

aacatgaagg taataaactt taacgttcca ctcaaaaaga caaaaaccaa acaacgaaaa    120 

ataagaaatt aaccagaaag ctatagcttg ttttcttact cagaaaaaaa gtataactga    180 

taaggtacaa tttctgtaac tggatatttt tcaaaattat aaggctttta gttctaaaag    240 

tataaagaac tgtgatgcac ttctagtcaa cctaatcttg ctagaagctt tatcaacact    300 

gacagtctca atactttctc ttttgctatt atatagtc                            338 

 
           
             75  
             334  
             DNA  
             Homo sapiens  
             
               misc_feature  
               265  
               n = A,T,C or G  
             
           
            75 

agcggccgcg gcggagcagc aacagttcta cctgctcctg ggaaacctgc tcagccccga     60 

caatgtggtc cggaaacagg cagaggaaac ctatgagaat atcccaggcc agtcaaagat    120 

cacattcctc ttacaagcca tcagaaatac aacagctgct gaagaggcta gacaaatggc    180 

cgccgttctc ctaagacgtc tcttgtcctc tgcatttgat ggaagtctat ccagcacttc    240 

cctcttgatg ttcagactgc catcnagagt gagctactca tgaattattc agatggaaac    300 

acaatctagc atgaggaaaa aaggtttgtg atat                                334 

 
           
             76  
             248  
             DNA  
             Homo sapiens  
             
               misc_feature  
               32,33  
               n = A,T,C or G  
             
           
            76 

gataggcata aacgtgttta ttaagtgaaa cnnatccttt aaaaataaaa aagggaagcc     60 

tgtatataaa tgaagttgtg gattcaacta gccagaattt attctgactt gcaccaaacc    120 

acacaaaatc ttttaaaagt ctagttagtc gtagtctaaa tggacactcc agagtctgtt    180 

cttgaattcc attgcaagag ctccaacttc ctactttcag aagggatggg gatcaagatg    240 

agggttgt                                                             248 

 
           
             77  
             515  
             DNA  
             Homo sapiens  
             
               misc_feature  
               395,476  
               n = A,T,C or G  
             
           
            77 

atgtagaaac agcatcaagc tgtttctctc taccgtcttt gatagaaata aaaataaaaa     60 

taaaaagttg aattgcagaa aagctaagag gtttttagtt tttgtttttt gttttccttc    120 

caccagtcaa ttattggaaa ggatttagtg agtctggttt attttagctt caatctgggt    180 

ttgtacacaa gcaaaaagca aatgttgaat tttcaggtag accttcatgc agacatgcaa    240 

aaccaactgt ctcggtggtg aggagccatg gggagctctc cgaagggctt tccaggcagt    300 

gggctaatgg gcaaaatgac tactcagtgg ccctgctgac cgatggtaac ggtgtgccaa    360 

ggatatctat cagcccatct gagaatatga aacanagtgc tgagattcta cttacctaag    420 

taacaaagaa accgtaagca acacgactga cagccagaag ggaacactgg aatggngggg    480 

tgaatggtgt cctgattagc accccccaat ctcgc                               515 

 
           
             78  
             532  
             DNA  
             Homo sapiens  
           
            78 

cctgttgtta tatagtttat tactgtcata gctaagaaaa ggcagtcgat ttcaacataa     60 

tccatatcta tgttcaaatt ctcaaactat aggatatcta tgtttcaaat tgtaatttat    120 

aacctggtaa gtattctaaa caaaatattg acaatccatt agctgaccta aaatcttatg    180 

aagctgtatc atcagtttaa caaatacaca cgactttagc aaaagtatat acagatagta    240 

tttataatac ttataataca ggcatggact aaaaaataca gataaaattg gagcaaatta    300 

aaagaggagt tgcattcaaa atattttttc catttgatat cattagaatt acaaaagcag    360 

taataaaaaa atctaatgtt aaggcaatga caaataacaa agataacagt tgcccaagga    420 

gcgaggggtt gggaggtgaa tgcacaatca aggaggggca caaaacagcc ttcaggttaa    480 

tttgttttat taagggggga gtcattggta gatagtcttt acatcttttt at            532 

 
           
             79  
             431  
             DNA  
             Homo sapiens  
           
            79 

gggataagca aaatgagtcc aacctttatt ctgataatag ccagtaaatt tgcaaagaga     60 

ggagacaaac tgtaattgta tacataaaaa cacctagtcc cactttaaaa ttttaatatc    120 

tatatatagt actgtattta atttttaaag atgaagacag caaaaatatt cacattaaaa    180 

tatcttacag aaatcattat tcttctattc aagaaaacca attatactaa gttaacaggg    240 

aaaatttaac agaggaaatt ctccttggga cacttattga actgaggatt tcacttcata    300 

gtttaaaaaa gtaaacaggt ctcaggtgtc tttttcatgg gtaggtcacc ttatcaatct    360 

gaattacagt tcatgggtaa agctaacttt ttttgtgtga aataagttaa taatgccaat    420 

tcagtttctt g                                                         431 

 
           
             80  
             431  
             DNA  
             Homo sapiens  
             
               misc_feature  
               361,431  
               n = A,T,C or G  
             
           
            80 

acaaaccttc cgggggttgc ctgagtggct gctctcggaa aagcggatcc taaataaagc     60 

gggagggtta tagggcgacg tcgaggagag gacaggtctc gagtcactgc tacagtttca    120 

ggtcactggg ctccgcagca gatcgtgttt tctcccgtgg ctcgagagct gcgctggttt    180 

ctcatgcaaa ctcagagccg agctaatgac atgagcaact tttactttta cacaagatga    240 

gcacgcgtgc cgaggcgctg ggcggcggct gtgtgagttg gtggcccaga cgaacagctt    300 

gtgcgagact ctgggcattt cggtttctag atacaagatt tgcttaaatg tcacagtcca    360 

nagaagtgga tttcagtcat tgtagctact ggatgcacac aaagtaaaaa aaaaaaaact    420 

tcacttgccg n                                                         431 

 
           
             81  
             471  
             DNA  
             Homo sapiens  
           
            81 

aaggtcagat attgtttaac acttgaaatt ccaaagagaa aaaatattcc caatgagtgc     60 

tctgtttcct atagagtaat tgctgaaata aaggaacaca gaaaacaagg cttctgccag    120 

ttgtcactta caaaaacata cagaggatca taatctagag acatggctaa ggcctcaggt    180 

ggtttcatgc tcaagattga tgttttgcca gagagctgag ttgtggagtc ctgtttcgga    240 

agggctgtga tggtggtgac ttcatcctca gctccttgct ttagggctcg ggcaagcttt    300 

tgaggtctgt aacttgttga agacttgtgg acagagaatg gctgatatct cttaattttg    360 

tacagttgag gaacctgcag attgaagaag gaataactct gcttgatttg aacttctgaa    420 

gacttaattg ggaccagtcc aaggccatca ggagccaact cgttggagtc c             471 

 
           
             82  
             450  
             DNA  
             Homo sapiens  
           
            82 

tgtcaatttt tgcaaatcaa agtgtatcat ttctccaatt ctactgatgc cagtttccaa     60 

gtccaattac tttttctacc ttctaatttt tcttaatttc taagccaata tgttaaaaac    120 

tattcttttg gctttcacaa tgttgcatta tcctaactgc ctctgatatc ttcaacaatt    180 

catttggtct ttaatgaaac tctttccatg taatgctctt tattaaatgt agatgtttcc    240 

ttaagaatga atctgcacca gccctttgct cttctccatg atttcaccta ctctcacaat    300 

ggtgatgggc attcccatgg ccctgacagc ttactgtatc tctttagcct gatctctccc    360 

tagaaatata atgttcatct gtgtttgtct gatgaggact gcctgatagc tgccaaatca    420 

acaaggataa aaccagaatt cacattccct                                     450 

 
           
             83  
             540  
             DNA  
             Homo sapiens  
           
            83 

ttatacaaaa gcatttaaca agcttaaaaa atgaaactca atgaaaaaaa aaagaaggtt     60 

tgaacacagt caaataacct gagaagtgac agatggaaaa gcaacagaat gcaagcacct    120 

tgtaaggtct gtaatctttg gatttactgt gaaaagtttc agaacatcat agactcttac    180 

tgccacattg tccatagacc ctggaaaata acagtgaaat tcatatgtat acacatatat    240 

atgaatacac actcatgcat gcacactgtc ttcacacacc cctcctcacc acttaaccgg    300 

agttacataa atgcttctca gatatgtcat tgcatttgtt tgttttctgc atctcaacta    360 

agttcagcgg cttgcgcctg tgacattaat tatgcaagat tcaaacaacc aagcaggcac    420 

attttggggg tgagttttaa gaaatctgtg acctgaaaga aattctgtgg ggactgtctg    480 

ggttatccag tttattccgt gattatattc tgtttttagg tcttgaccta tttttaagct    540 

 
           
             84  
             559  
             DNA  
             Homo sapiens  
             
               misc_feature  
               493,499,506,517,537,550,559  
               n = A,T,C or G  
             
           
            84 

gttgttgctg ctgtttttac tcggacaatg cttattttac agcggaattg acaaataaag     60 

ccttatttta cacatccgaa gaaacaccat cacaggaggt ttgtaggtcg gctgtgtgct    120 

ttccaaaaca gcaaaataga ttcttcccat ccaaccccct ttcctcttgt agagtagggt    180 

gtggctcgtg gggcttcgtc tctctgcagg cacagaaact ggcagacctg gtccctcctg    240 

agcgggccct gctcaaggga atggtgccag attttgaaca caggtaaaca ggctccttca    300 

taacaacact gtgcatttct gtgtcatttt gtttattgct cactgagttg ttgccacctc    360 

agctcttggt ggaaaacagt gggtgtccag aaattgctga cacaagaaga tggattgcct    420 

atggtccgtt agggacacag ggcagcccca gccagatccc actggtccat gcagggcatc    480 

gcagtagaaa ctnaacgtnc cacttngtaa caggctncaa gacaccaatt ccggcancat    540 

gggaaagaan taaaccttn                                                 559 

 
           
             85  
             2466  
             DNA  
             Homo sapiens  
           
            85 

agttggtccg agctgccgaa aggtctggtc gcagagacag gaacgtgtaa tcctcagcgt     60 

gctccagccc acagcttcgc tctactgctc ggcagggcag ctggcctctg ggcaccggcg    120 

gcccctctgc ctcgcggaaa agcctgatga agtcctccga tattgatcag gatttattca    180 

cagacagtta ctgcaaggtg tgcagtgcac agctgatctc cgaatcgcag cgtgtggccc    240 

actacgagag tcgaaaacat gcaagcaaag tccgactgta ttacatgctt caccccaggg    300 

atggagggtg tcctgccaag aggctccggt cagaaaatgg aagtgatgcc gacatggtgg    360 

ataagaacaa gtgctgcaca ctctgcaaca tgtcattcac ttcagcggtg gtggccgatt    420 

cccattatca aggcaaaatc cacgccaaga ggttaaaact cttgctagga gagaagaccc    480 

cattaaagac cacagcaaca cccctgagcc cacttaagcc cccacggatg gacactgctc    540 

cggtggtcgc atctccctat caaagaagag attcagacag atactgtggg ctctgtgcag    600 

cctggtttaa taaccctctg atggcccagc aacattatga tggcaagaaa cacaaaaaga    660 

atgcggcaag agttgctttg ttagaacaac tggggacaac cctggatatg ggggaactga    720 

gaggtctgag gcgcaattac agatgtacca tctgcagtgt ctccctaaac tcaatagaac    780 

agtatcatgc ccatctgaaa ggatctaaac accagaccaa cctgaagaat aagtagtgaa    840 

agcatcaatc aagacataag aacaaaacat tagcatttct ctgccgtgga gaattgctta    900 

tcaaccacca gaggaggctt ctttcttgaa caataaacat ttcttataag gattcacaga    960 

ttcacatacg actgatcttg atttttggaa atgaatgagg tttctttttt ctttttcctt   1020 

tttttaattt tggggtaagt tatgatattt ggatggattt ttaaattctt tcctgataac   1080 

atatttagca catgttctaa attataatcc tatagcaaac agttggagca ttattcaaac   1140 

tgaaagtgga aaaatttaaa tttccaattt attctagatt tcctcagagc ataattattc   1200 

tgttaaatcc tcaatgagtg tgatgtaaac cacctctatc cagaaatata cattcttttc   1260 

tcatcatgtt ggacacagtt gagggtgaca tgcacagaac tggaacagat cactattagt   1320 

ggaaaatacc aaatggacaa ataaatacca gtcgttttct ccgttctcca agcacaggag   1380 

ccaggtttac catctgaaca atgaagacga agggagtaaa taaaggaaga attctcatct   1440 

tttttcctga tcattcaaag aacagtttct caaggttaag ccaagtcctc cttgcaagtt   1500 

gccaaataat agcttaggaa aagaattagt ctgcctgcat gatgatcttc ttaggcaaaa   1560 

acgtcttcac agcccttgac cttggtgaat ttttttcccc aaaagcatcc aaaagaagaa   1620 

ttataaaccc cagaacgaga tggaaataaa caagtatttt ttttttatga tgtttggcct   1680 

gaactgtggg ctttaattgg gggatactga tcgtttggaa agaagtgaga aaattctgaa   1740 

gaaatggcgg ccttgggcta ggcggggtcc cctatttctt ctgtttctca ctgaagtcct   1800 

actgctgagc caagactcag tcactctgga aagagcatga ccgataaaga aaacagttcc   1860 

tttctgatgg ggagcgtctg agtgcagatc atgaggctct ttctctaggt ttaattcttt   1920 

tccatggtga ccggacttgg tgtcttgtag cctggttacg aagtgggacg ttgagcttct   1980 

actgacgatg ccctgcatgg accagctggg atctggctgg ggctgccctg tgtccctaac   2040 

gaccataggc aatccatctt cttgtgtcag caatttctgg acacccactg ttttccacca   2100 

agagctgagg tggcaacaac tcagtgagca ataaacaaaa tgacacagaa atgcacagtg   2160 

ttgttatgaa ggagcctgtt tacctgtgtt caaaatctgg caccattccc ttgagcaggg   2220 

cccgctcagg agggaccagg tctgccagtt tctgtgcctg cagagagacg aagccccacg   2280 

agccacaccc tactctacaa gaggaaaggg ggttggatgg gaagaatcta ttttgctgtt   2340 

ttggaaagca cacagccgac ctacaaacct cctgtgatgg tgtttcttcg gatgtgtaaa   2400 

ataaggcttt atttgtcaat tccgctgtaa aataagcatt gtccgagtaa aaacagcagc   2460 

aacaac                                                              2466 

 
           
             86  
             408  
             DNA  
             Homo sapiens  
           
            86 

ttttttggca tttaagtttt tcaccaattt attgctaaga ggaaacatat aataatatgc     60 

tatagggtca taaaacccac tttgcagcta tagaagcaag ttctgcctgt gcctgtgtat    120 

gtgtatgtat gacagtggac atgtaagtgt gaaactttaa acactattac agtaagaagt    180 

cttttgttga acttttgtta gtttgagagg ctgcaatgat ttttctcctt tcaaaatgct    240 

gaaatagaac tcatcatttt gcttttcaaa ttagcaacag gtagctggtt tggaaggctg    300 

gagattgatt tctctccagc tagcaagtcg tggggtcagg tcactgaagc atgtgggtga    360 

tatgctgaac caccaacttg gcaaatattg aactatttta agtgcatc                 408 

 
           
             87  
             431  
             DNA  
             Homo sapiens  
             
               misc_feature  
               361,431  
               n = A,T,C or G  
             
           
            87 

acaaaccttc cgggggttgc ctgagtggct gctctcggaa aagcggatcc taaataaagc     60 

gggagggtta tagggcgacg tcgaggagag gacaggtctc gagtcactgc tacagtttca    120 

ggtcactggg ctccgcagca gatcgtgttt tctcccgtgg ctcgagagct gcgctggttt    180 

ctcatgcaaa ctcagagccg agctaatgac atgagcaact tttactttta cacaagatga    240 

gcacgcgtgc cgaggcgctg ggcggcggct gtgtgagttg gtggcccaga cgaacagctt    300 

gtgcgagact ctgggcattt cggtttctag atacaagatt tgcttaaatg tcacagtcca    360 

nagaagtgga tttcagtcat tgtagctact ggatgcacac aaagtaaaaa aaaaaaaact    420 

tcacttgccg n                                                         431 

 
           
             88  
             385  
             DNA  
             Homo sapiens  
           
            88 

gaatattcag tccacaaatt ggcagacaat gagatttaag ccccctcctc caaactcaga     60 

cattggatgg agagtagaat ttcgacccat ggaggtgcaa ttaacagact ttgagaactc    120 

tgcctatgtg gtgtttgtgg tactgctcac cagagtgatc ctttcctaca aattggattt    180 

tctcattcca ctgtcaaagg ttgatgagaa catgaaggta gcacagaaaa gagatgctgt    240 

cttgcaggga atgttttatt tcaggaaaga tatttgcaaa ggtggcaatg cagtggtgga    300 

tggttgtggc aaggcccaga acagcacgga gctcgctgca gaggagtaca ccctcatgag    360 

catagacacc atcatcaatg ggaac                                          385 

 
           
             89  
             272  
             DNA  
             Homo sapiens  
           
            89 

tctttaaaat acatacgaat gtaaagagaa aatggccaaa acctcaaaac tacgattgtt     60 

gaaaacaata ttaaaaggac acaatctaaa atcatgctac aaaaatagtg ttatcttgtt    120 

taactaaatg tacatctttt tttccaattc catgattgac aagagtgctt atgcgacgca    180 

tggaaggcac cagaggtgaa gtgattattt gccttaaaat atacaaagaa ttgcctactt    240 

tgaaaaagaa atagtcatac ttgtaaatga at                                  272 

 
           
             90  
             504  
             DNA  
             Homo sapiens  
           
            90 

gaagcagttt attaccttaa agcatttagc aaacctaatg tctgacctaa tttcaaccaa     60 

atgtctttat tttaccaata atcttcaaaa ctcttgattt cccaaagcct actaaagtca    120 

tgctgtcaca ggccattaga cagcatgagc agggcaggaa agggctcttc tcccacccac    180 

caggaatgtt gggtgatggc tcagcagtta tcacattgcc tctctaaaag tcatacattg    240 

gcacctaggg tcagggagac gccatttcct gatggtccac acctattgca ctaaagtgtt    300 

aattgaatgc agatgccagg gagatgcaac ttcccaggca aatgcattaa gagacaaaac    360 

ggcagagtat gacctttccg tggcactcca tgggaaaagg gaagaaagcc ttgggtgggc    420 

atgtgtacaa cttcctaaac acactgcatg tgctcacctc ccaaggatag ggagggcact    480 

gtgcatgcgg gcagctcacc ctaa                                           504 

 
           
             91  
             467  
             DNA  
             Homo sapiens  
           
            91 

tttttttttt ttttttttgc tttctcaaca aatagtttac tcggtggaac ctaacagaac     60 

taatatttct ttctgtccgt aaataaaaat agatcatgct tgaatgtgct actttgcccg    120 

aactccccaa gtcttcccgc atcttcagtt cctccccctc caacctggtg tttatcagga    180 

gaggggaaag agcatttctt gcctggcagg aactcaagac ctagaagaaa gagggcctac    240 

cctgccaagg aaacgacctt ccccttcctc gcctctgctc ctcttcccgt ttcctgtctt    300 

ttccttcttt tctcctgggg tttccttctc ccgttaacta tggggacaga cacagctatt    360 

cacaagtccg tctgggcagc acactccgag gtaaggcacg aaggtcagga gacaggttcc    420 

cgtgccccaa atcctggaga agatgagtta aagctcttcg cttcgat                  467 

 
           
             92  
             229  
             PRT  
             Homo sapiens  
           
            92 

Met Lys Ser Ser Asp Ile Asp Gln Asp Leu Phe Thr Asp Ser Tyr Cys 
                  5                  10                  15 

Lys Val Cys Ser Ala Gln Leu Ile Ser Glu Ser Gln Arg Val Ala His 
             20                  25                  30 

Tyr Glu Ser Arg Lys His Ala Ser Lys Val Arg Leu Tyr Tyr Met Leu 
         35                  40                  45 

His Pro Arg Asp Gly Gly Cys Pro Ala Lys Arg Leu Arg Ser Glu Asn 
     50                  55                  60 

Gly Ser Asp Ala Asp Met Val Asp Lys Asn Lys Cys Cys Thr Leu Cys 
 65                  70                  75                  80 

Asn Met Ser Phe Thr Ser Ala Val Val Ala Asp Ser His Tyr Gln Gly 
                 85                  90                  95 

Lys Ile His Ala Lys Arg Leu Lys Leu Leu Leu Gly Glu Lys Thr Pro 
            100                 105                 110 

Leu Lys Thr Thr Ala Thr Pro Leu Ser Pro Leu Lys Pro Pro Arg Met 
        115                 120                 125 

Asp Thr Ala Pro Val Val Ala Ser Pro Tyr Gln Arg Arg Asp Ser Asp 
    130                 135                 140 

Arg Tyr Cys Gly Leu Cys Ala Ala Trp Phe Asn Asn Pro Leu Met Ala 
145                 150                 155                 160 

Gln Gln His Tyr Asp Gly Lys Lys His Lys Lys Asn Ala Ala Arg Val 
                165                 170                 175 

Ala Leu Leu Glu Gln Leu Gly Thr Thr Leu Asp Met Gly Glu Leu Arg 
            180                 185                 190 

Gly Leu Arg Arg Asn Tyr Arg Cys Thr Ile Cys Ser Val Ser Leu Asn 
        195                 200                 205 

Ser Ile Glu Gln Tyr His Ala His Leu Lys Gly Ser Lys His Gln Thr 
    210                 215                 220 

Asn Leu Lys Asn Lys 
225 

 
           
             93  
             2327  
             DNA  
             Homo sapiens  
           
            93 

gggagcgaaa accaacgtgt tcggtgacag accccagcgc cgactgagcc tctaaagcga     60 

cttcagctct gccccaccaa caccaccgcg cgcccgggaa cagccgctcc gggaagaaac    120 

ctgaggggac tgcggggggc acgagggaca gctgagggaa gggaggacgc gagagaaaca    180 

gcgcaagcac gctgagggcc gggggttgcc aggagagggg cccgcggacc cgcagagcgg    240 

aggaaggtcc gggagaaaag gggcgggacg gaggagaatc cgggatcgcc tggcagaaaa    300 

agagaaggga gtttctgaat cctgggaaga ggaggcgtgg gtagggatgc ttagcccgag    360 

atccgacagc agggaaccgg agcgctccgg gggaggggct taatgctggg gaagggatgt    420 

cttaaaagag gagaagcttt aaattagacg atcggagaag gctgagggaa ttgctatgaa    480 

ggggcgggag ctgaagtgta gaggactcct ttagacagca gaaagggaaa gccgttgaga    540 

agttcccttc aaactccacc tgcctcctct ccaattcaaa ctccactccc ttctccaaaa    600 

gttaaaagga aagccaagtt tgccacgctc ccctgttcct actcaataaa tacttcttct    660 

actccgccac cgggaaaaca gaaaaaaaaa actaatttcc ttcccaatat taggacttag    720 

aaaagctcta ggtcccgcaa cttgaatttt agcctagggg aatcaaaata gtaggagcat    780 

tactcttgtt tcctttttca aaatcccaca cctcatcctt cctgcgacgc catgtctacc    840 

aacatttgta gtttcaagga caggtgcgtg tccatcctgt gttgcaaatt ctgtaaacaa    900 

gtgctcagct ctaggggaat gaaggctgtt ttgctggctg atactgaaat agaccttttc    960 

tctacagaca tccctcctac caacgcagtg gacttcactg gaagatgcta tttcaccaaa   1020 

atctgcaaat gtaaactgaa ggacatcgca tgtttaaaat gtgggaacat tgtaggttat   1080 

catgtgattg ttccatgtag ttcctgtctt ctttcctgca acaacggaca cttctggatg   1140 

tttcacagcc aggcagttta tgatattaac agactagact ccacaggtgt aaacgtccta   1200 

ctttggggca acttgccaga gatagaagag agtacagatg aagatgtgtt aaatatctca   1260 

gcagaggagt gtattagata aatggaatta tgatatatat gatatacaaa cttttttcta   1320 

tttaaaaata tattaatgga tcaactttaa aattgttagt tgccagtgat cttttttgga   1380 

aaacaaaaat ggggcatttg ttgatttatt tattttccgt ctctaattag ttacctcagt   1440 

ttgattgaag ccagtggagt tgtgcttttc ctctacttct acttcctctc ccccaccttt   1500 

ttctgcccag tgtaggtgta ttcttaaatt cagacgggaa gattctttca catatcactc   1560 

agttacctcc caatctgggg gagtttttct tacaacttga taccagatac cattaatttt   1620 

acattcctga ataaaggcct agtacccacg catatttcaa ccatgcatat atcaagttca   1680 

actgagtttt aataggggat taaaaaaaca agctgttagg tttccatggg cactggttct   1740 

cataggttct attggtgata actgctttaa catggagcaa gagtttgtga atcaggaaat   1800 

agaataaatt aaaatttaaa atatatagag gaatcctctt gattgctcag catgatgtta   1860 

gataaatgag tttgtcagaa aatatcagta tacgctgttt accaatgtta tttatttaca   1920 

ttcttctaaa gccattatgg atattgtatt atgagagcta aacctaaata agttatcctg   1980 

ttccctagga ccttctctgt aaatagtgaa ttttagacga gtagtctgtc ctaaatctta   2040 

aatagaaaaa aaaactaaag cgatttgctt aagccattgt acattataaa gagctgtttt   2100 

gttttgcttt gctttgcttt gttttgtttt ttttaaagct gcattcagag ccacaaagga   2160 

ataggaaagt agggtagtgt tggattctgg ttttatgtaa ctctaaaata aatgtatctc   2220 

tttaatatct cagttgtagg gattttgtca ataccaaagc agactgagtt gtggttttgt   2280 

aaataaagtt ttttctaaaa atgaaaaaaa aagaaaaaaa aaaaaaa                 2327 

 
           
             94  
             2370  
             DNA  
             Homo sapiens  
             
               misc_feature  
               741,1195,1683,2360  
               n = A,T,C or G  
             
           
            94 

gggagcgaaa accaacgtgt tcggtgacag accccagcgc cgactgagcc tctaaagcga     60 

cttcagctct gccccaccaa caccaccgcg cgcccgggaa cagccgctcc gggaagaaac    120 

ctgaggggac tgcggggggc acgagggaca gctgagggaa gggaggacgc gagagaaaca    180 

gcgcaagcac gctgagggcc gggggttgcc aggagagggg cccgcggacc cgcagagcgg    240 

aggaaggtcc gggagaaaag gggcgggacg gaggagaatc cgggatcgcc tggcagaaaa    300 

agagaaggga gtttctgaat cctgggaaga ggaggcgtgg gtagggatgc ttagcccgag    360 

atccgacagc agggaaccgg agcgctccgg gggaggggct taatgctggg gaagggatgt    420 

cttaaaagag gagaagcttt aaattagacg atcggagaag gctgagggaa ttgctatgaa    480 

ggggcgggag ctgaagtgta gaggactcct ttagacagca gaaagggaaa gccgttgaga    540 

agttcccttc aaactccacc tgcctcctct ccaattcaaa ctccactccc ttctccaaaa    600 

gttaaaagga aagccaagtt tgccacgctc ccctgttcct actcaataaa tacttcttct    660 

actccgccac cgggaaaaca gaaaaaaaaa actaatttcc ttcccaatat taggacttag    720 

aaaagctcta ggtcccgcaa yttgaatttt agcctagggg aatcaaaata gtaggagcat    780 

tactcttgtt tcctttttca aaatcccaca cctcatcctt cctgcgacgc catgtctacc    840 

aacatttgta gtttcaagga caggtgcgtg tccatcctgt gttgcaaatt ctgtaaacaa    900 

gtgctcagct ctaggggaat gaaggctgtt ttgctggctg atactgaaat agaccttttc    960 

tctacagaca tccctcctac caacgcagtg gacttcactg gaagatgcta tttcaccaaa   1020 

atctgcaaat gtaaactgaa ggacatcgca tgtttaaaat gtgggaacat tgtaggttat   1080 

catgtgattg ttccatgtag ttcctgtctt ctttcctgca acaacggaca cttctggatg   1140 

tttcacagcc aggcagttta tgatattaac agactagact ccacaggtgt aaacrtccta   1200 

ctttggggca acttgccaga gatagaagag agtacagatg aagatgtgtt aaatatctca   1260 

gcagaggagt gtattagata aatggaatta tgatatatat gatatacaaa cttttttcta   1320 

tttaaaaata tattaatgga tcaactttaa aattgttagt tgccagtgat cttttttgga   1380 

aaacaaaaat ggggcatttg ttgatttatt tattttccgt ctctaattag ttacctcagt   1440 

ttgattgaag ccagtggagt tgtgcttttc ctctacttct acttcctctc ccccaccttt   1500 

ttctgcccag tgtaggtgta ttcttaaatt cagacgggaa gattctttca catatcactc   1560 

agttacctcc caatctgggg gagtttttct tacaacttga taccagatac cattaatttt   1620 

acattcctga ataaaggcct agtacccacg catatttcaa ccatgcatat atcaagttca   1680 

acygagtttt aataggggat taaaaaaaca agctgttagg tttccatggg cactggttct   1740 

cataggttct attggtgata actgctttaa catggagcaa gagtttgtga atcaggaaat   1800 

agaataaatt aaaatttaaa atatatagag gaatcctctt gattgctcag catgatgtta   1860 

gataaatgag tttgtcagaa aatatcagta tacgctgttt accaatgtta tttatttaca   1920 

ttcttctaaa gccattatgg atattgtatt atgagagcta aacctaaata agttatcctg   1980 

ttccctagga ccttctctgt aaatagtgaa ttttagacga gtagtctgtc ctaaatctta   2040 

aatagaaaaa aaaactaaag cgatttgctt aagccattgt acattataaa gagctgtttt   2100 

gttttgcttt gctttgcttt gttttgtttt ttttaaagct gcattcagag ccacaaagga   2160 

ataggaaagt agggtagtgt tggattctgg ttttatgtaa ctctacccta ctttcctatt   2220 

cctttgtgtc ctgtaacttt ttttacctat caatatgagt tgctgtgctt cagtgtgtat   2280 

tttttaagtt gctgggcatt acacttacca attaaagaat tttggaaatt caaaaaaaaa   2340 

aaaaaaaaaa aaaaaaaaam aaaaaaaaaa                                    2370 

 
           
             95  
             450  
             DNA  
             Homo sapiens  
           
            95 

atgtctacca acatttgtag tttcaaggac aggtgcgtgt ccatcctgtg ttgcaaattc     60 

tgtaaacaag tgctcagctc taggggaatg aaggctgttt tgctggctga tactgaaata    120 

gaccttttct ctacagacat ccctcctacc aacgcagtgg acttcactgg aagatgctat    180 

ttcaccaaaa tctgcaaatg taaactgaag gacatcgcat gtttaaaatg tgggaacatt    240 

gtaggttatc atgtgattgt tccatgtagt tcctgtcttc tttcctgcaa caacggacac    300 

ttctggatgt ttcacagcca ggcagtttat gatattaaca gactagactc cacaggtgta    360 

aacgtcctac tttggggcaa cttgccagag atagaagaga gtacagatga agatgtgtta    420 

aatatctcag cagaggagtg tattagataa                                     450 

 
           
             96  
             149  
             PRT  
             Homo sapiens  
           
            96 

Met Ser Thr Asn Ile Cys Ser Phe Lys Asp Arg Cys Val Ser Ile Leu 
                  5                  10                  15 

Cys Cys Lys Phe Cys Lys Gln Val Leu Ser Ser Arg Gly Met Lys Ala 
             20                  25                  30 

Val Leu Leu Ala Asp Thr Glu Ile Asp Leu Phe Ser Thr Asp Ile Pro 
         35                  40                  45 

Pro Thr Asn Ala Val Asp Phe Thr Gly Arg Cys Tyr Phe Thr Lys Ile 
     50                  55                  60 

Cys Lys Cys Lys Leu Lys Asp Ile Ala Cys Leu Lys Cys Gly Asn Ile 
 65                  70                  75                  80 

Val Gly Tyr His Val Ile Val Pro Cys Ser Ser Cys Leu Leu Ser Cys 
                 85                  90                  95 

Asn Asn Gly His Phe Trp Met Phe His Ser Gln Ala Val Tyr Asp Ile 
            100                 105                 110 

Asn Arg Leu Asp Ser Thr Gly Val Asn Val Leu Leu Trp Gly Asn Leu 
        115                 120                 125 

Pro Glu Ile Glu Glu Ser Thr Asp Glu Asp Val Leu Asn Ile Ser Ala 
    130                 135                 140 

Glu Glu Cys Ile Arg 
145