LOCUS HSPA5-pGEn3-DEST 7917 bp DNA circular 26-JAN-2013 FEATURES Location/Qualifiers CDS 129..1108 /dnas_title="AmpR" /label=AmpR /vntifkey="4" rep_origin 1137..1804 /dnas_title="Col E1 origin" /label=Col E1 origin /vntifkey="33" misc_feature 1998..2127 /note="5'ITR" misc_feature 2208..2790 /note="CMV Promoter" misc_feature 2828..2960 /note="Artificial Intron" misc_recomb complement(4541..4569) /label=attB1 /note="attB1 site" /note="recognized as attB1" misc_feature 6545..7086 /note="WPRE" misc_recomb 6507..6529 /label=attB2 /note="attB2 site" /note="recognized as attB2" misc_feature complement(7093..7307) /note="BGH pA" misc_feature complement(7395..7524) /note="3'-ITR" misc_feature 2982..3056 /note="TCM signal sequence" misc_feature 3060..3083 /note="His tag" misc_feature 3084..3134 /note="AviTag" misc_feature 3135..3830 /note="Superfolder GFP" misc_feature 3837..4532 /note="Fc domain" misc_feature 2976..2981 /note="Kozak seq" misc_feature 4570..4592 /note="TEV site" source 4593..6500 /dnas_title="HSPA5-pGEn3-DEST" /organism="Homo sapiens" /mol_type="mRNA" /db_xref="taxon:9606" /clone="MGC:31939 IMAGE:5020098" /tissue_type="Muscle, rhabdomyosarcoma" /clone_lib="NIH_MGC_17" /lab_host="DH10B-R" /note="Vector: pOTB7" gene 4593..6500 /dnas_title="HSPA5" /gene="HSPA5" /gene_synonym="BIP" /gene_synonym="MIF2" /db_xref="GeneID:3309" /db_xref="HGNC:5238" /db_xref="MIM:138120" CDS 4593..6500 /dnas_title="heat shock 70kDa protein 5 (glucose-regulated protein, 78kDa)" /gene="HSPA5" /gene_synonym="BIP" /gene_synonym="MIF2" /codon_start=1 /product="heat shock 70kDa protein 5 (glucose-regulated protein, 78kDa)" /protein_id="AAH20235.1" /db_xref="GI:18044381" /db_xref="GeneID:3309" /db_xref="HGNC:5238" /db_xref="MIM:138120" /translation="MKLSLVAAMLLLLSAARAEEEDKKEDVGTVVGIDLGTTYSCVGV FKNGRVEIIANDQGNRITPSYVAFTPEGERLIGDAAKNQLTSNPENTVFDAKRLIGRT WNDPSVQQDIKFLPFKVVEKKTKPYIQVDIGGGQTKTFAPEEISAMVLTKMKETAEAY LGKKVTHAVVTVPAYFNDAQRQATKDAGTIAGLNVMRIINEPTAAAIAYGLDKREGEK NILVFDLGGGTFDVSLLTIDNGVFEVVATNGDTHLGGEDFDQRVMEHFIKLYKKKTGK DVRKDNRAVQKLRREVEKAKRALSSQHQARIEIESFYEGEDFSETLTRAKFEELNMDL FRSTMKPVQKVLEDSDLKKSDIDEIVLVGGSTRIPKIQQLVKEFFNGKEPSRGINPDE AVAYGAAVQAGVLSGDQDTGDLVLLDVCPLTLGIETVGGVMTKLIPRNTVVPTKKSQI FSTASDNQPTVTIKVYEGERPLTKDNHLLGTFDLTGIPPAPRGVPQIEVTFEIDVNGI LRVTAEDKGTGNKNKITITNDQNRLTPEEIERMVNDAEKFAEEDKKLKERIDTRNELE SYAYSLKNQIGDKEKLGGKLSSEDKETMEKAVEEKIEWLESHQDADIEDFKAKKKELE EIVQPIISKLYGSAGPPPTGEEDTAEKDEL" misc_feature 4593..6500 /note="HSPA5 coding region" vector join(6515..7917,1..4559) /source="pGEn3-DEST" /type="Destination Vector" clone 4560..6514 /source="HSPA5-Nterm-TEVfusion-pDONR221" /type="Entry Clone" ORIGIN 1 GTGGCACTTT TCGGGGAAAT GTGCGCGGAA CCCCTATTTG TTTATTTTTC TAAATACATT 61 CAAATATGTA TCCGCTCATG AGACAATAAC CCTGATAAAT GCTTCAATAA TATTGAAAAA 121 GGAAGAGTAT GAGTATTCAA CATTTCCGTG TCGCCCTTAT TCCCTTTTTT GCGGCATTTT 181 GCCTTCCTGT TTTTGCTCAC CCAGAAACGC TGGTGAAAGT AAAAGATGCT GAAGATCAGT 241 TGGGTGCACG AGTGGGTTAC ATCGAACTGG ATCTCAACAG CGGTAAGATC CTTGAGAGTT 301 TTCGCCCCGA AGAACGTTTT CCAATGATGA GCACTTTTAA AGTTCTGCTA TGTGGCGCGG 361 TATTATCCCG TATTGACGCC GGGCAAGAGC AACTCGGTCG CCGCATACAC TATTCTCAGA 421 ATGACTTGGT TGAGTACTCA CCAGTCACAG AAAAGCATCT TACGGATGGC ATGACAGTAA 481 GAGAATTATG CAGTGCTGCC ATAACCATGA GTGATAACAC TGCGGCCAAC TTACTTCTGA 541 CAACGATCGG AGGACCGAAG GAGCTAACCG CTTTTTTGCA CAACATGGGG GATCATGTAA 601 CTCGCCTTGA TCGTTGGGAA CCGGAGCTGA ATGAAGCCAT ACCAAACGAC GAGCGTGACA 661 CCACGATGCC TGTAGCAATG GCAACAACGT TGCGCAAACT ATTAACTGGC GAACTACTTA 721 CTCTAGCTTC CCGGCAACAA TTAATAGACT GGATGGAGGC GGATAAAGTT GCAGGACCAC 781 TTCTGCGCTC GGCCCTTCCG GCTGGCTGGT TTATTGCTGA TAAATCTGGA GCCGGTGAGC 841 GTGGTTCTCG CGGTATCATT GCAGCACTGG GGCCAGATGG TAAGCCCTCC CGTATCGTAG 901 TTATCTACAC GACGGGGAGT CAGGCAACTA TGGATGAACG AAATAGACAG ATCGCTGAGA 961 TAGGTGCCTC ACTGATTAAG CATTGGTAAC TGTCAGACCA AGTTTACTCA TATATACTTT 1021 AGATTGATTT AAAACTTCAT TTTTAATTTA AAAGGATCTA GGTGAAGATC CTTTTTGATA 1081 ATCTCATGAC CAAAATCCCT TAACGTGAGT TTTCGTTCCA CTGAGCGTCA GACCCCGTAG 1141 AAAAGATCAA AGGATCTTCT TGAGATCCTT TTTTTCTGCG CGTAATCTGC TGCTTGCAAA 1201 CAAAAAAACC ACCGCTACCA GCGGTGGTTT GTTTGCCGGA TCAAGAGCTA CCAACTCTTT 1261 TTCCGAAGGT AACTGGCTTC AGCAGAGCGC AGATACCAAA TACTGTTCTT CTAGTGTAGC 1321 CGTAGTTAGG CCACCACTTC AAGAACTCTG TAGCACCGCC TACATACCTC GCTCTGCTAA 1381 TCCTGTTACC AGTGGCTGCT GCCAGTGGCG ATAAGTCGTG TCTTACCGGG TTGGACTCAA 1441 GACGATAGTT ACCGGATAAG GCGCAGCGGT CGGGCTGAAC GGGGGGTTCG TGCACACAGC 1501 CCAGCTTGGA GCGAACGACC TACACCGAAC TGAGATACCT ACAGCGTGAG CTATGAGAAA 1561 GCGCCACGCT TCCCGAAGGG AGAAAGGCGG ACAGGTATCC GGTAAGCGGC AGGGTCGGAA 1621 CAGGAGAGCG CACGAGGGAG CTTCCAGGGG GAAACGCCTG GTATCTTTAT AGTCCTGTCG 1681 GGTTTCGCCA CCTCTGACTT GAGCGTCGAT TTTTGTGATG CTCGTCAGGG GGGCGGAGCC 1741 TATGGAAAAA CGCCAGCAAC GCGGCCTTTT TACGGTTCCT GGCCTTTTGC TGGCCTTTTG 1801 CTCATTAGGC ACCCCAGGCT TTACCCGAAC GACCGAGCGC AGCGAGTCAG TGAGCGAGGA 1861 AGCGGAGAGC GCCCAATACG CAAGGAAACA GCTATGACCA TGTTAATGCA GCTGGCACGA 1921 CAGGTTTCCC GACTGGAAAG CGGGCAGTGA GCGGAAGGCC CATGAGGCCA GTTAATTAAG 1981 AGGTACCTTA ATTAAGGCTG CGCGCTCGCT CGCTCACTGA GGCCGCCCGG GCAAAGCCCG 2041 GGCGTCGGGC GACCTTTGGT CGCCCGGCCT CAGTGAGCGA GCGAGCGCGC AGAGAGGGAG 2101 TGGCCAACTC CATCACTAGG GGTTCCTTGT AGTTAATGAT TAACCCGCCA TGCTACTTAT 2161 CTACGTAGCC ATGCTCTAGG AAGATCGCAA TTCGCCCTTA AGCTAGCTAG TTATTAATAG 2221 TAATCAATTA CGGGGTCATT AGTTCATAGC CCATATATGG AGTTCCGCGT TACATAACTT 2281 ACGGTAAATG GCCCGCCTGG CTGACCGCCC AACGACCCCC GCCCATTGAC GTCAATAATG 2341 ACGTATGTTC CCATAGTAAC GCCAATAGGG ACTTTCCATT GACGTCAATG GGTGGAGTAT 2401 TTACGGTAAA CTGCCCACTT GGCAGTACAT CAAGTGTATC ATATGCCAAG TACGCCCCCT 2461 ATTGACGTCA ATGACGGTAA ATGGCCCGCC TGGCATTATG CCCAGTACAT GACCTTATGG 2521 GACTTTCCTA CTTGGCAGTA CATCTACGTA TTAGTCATCG CTATTACCAT GGTGATGCGG 2581 TTTTGGCAGT ACATCAATGG GCGTGGATAG CGGTTTGACT CACGGGGATT TCCAAGTCTC 2641 CACCCCATTG ACGTCAATGG GAGTTTGTTT TGGCACCAAA ATCAACGGGA CTTTCCAAAA 2701 TGTCGTAACA ACTCCGCCCC ATTGACGCAA ATGGGCGGTA GGCGTGTACG GTGGGAGGTC 2761 TATATAAGCA GAGCTGGTTT AGTGAACCGT CAGATCCTGC AGAAGTTGGT CGTGAGGCAC 2821 TGGGCAGGTA AGTATCAAGG TTACAAGACA GGTTTAAGGA GACCAATAGA AACTGGGCTT 2881 GTCGAGACAG AGAAGACTCT TGCGTTTCTG ATAGGCACCT ATTGGTCTTA CTGACATCCA 2941 CTTTGCCTTT CTCTCCACAG GTGTCCAgcg gccgcGCCAC cATGAGACTG CTGACCGCCC 3001 TGTTCGCCTA CTTCATCGTG GCCCTGATCC TGGCCTTCAG CGTGTCCGCC AAGAGCATGC 3061 ACCACCACCA TCACCATCAT CACATGAGCG GCCTGAACGA CATCTTCGAG GCCCAGAAAA 3121 TCGAGTGGCA CGAGATGAGC AAGGGCGAGG AACTGTTCAC CGGCGTGGTG CCCATCCTGG 3181 TGGAACTGGA CGGCGACGTG AACGGCCACA AGTTCTCTGT GCGGGGCGAG GGCGAGGGGG 3241 ACGCCACAAA TGGCAAGCTG ACCCTGAAGT TCATCTGCAC CACCGGAAAG CTGCCCGTGC 3301 CCTGGCCTAC CCTGGTCACA ACCCTGACCT ACGGCGTGCA GTGCTTCAGC AGATACCCCG 3361 ACCACATGAA GCGGCACGAT TTCTTCAAGA GCGCCATGCC CGAGGGCTAC GTGCAAGAAC 3421 GGACCATCAG CTTCAAGGAC GACGGCACCT ACAAGACCAG AGCCGAAGTG AAGTTCGAGG 3481 GCGACACCCT GGTCAACCGG ATCGAGCTGA AGGGCATCGA CTTCAAAGAG GACGGCAACA 3541 TCCTGGGCCA CAAGCTCGAG TACAACTTCA ACAGCCACAA CGTGTACATC ACCGCCGACA 3601 AGCAGAAGAA CGGCATCAAG GCCAACTTCA AGATCCGGCA CAACGTGGAA GATGGCAGCG 3661 TGCAGCTGGC CGACCACTAC CAGCAGAACA CCCCCATCGG CGACGGCCCC GTGCTGCTGC 3721 CCGACAATCA CTACCTGAGC ACCCAGAGCG TGCTGAGCAA GGACCCCAAC GAGAAGCGGG 3781 ACCACATGGT GCTGCTGGAA TTTGTGACCG CCGCTGGCAT CACCCACGGC GAATTCgagc 3841 ccaaatcttg tgacaaaact cacacatgcc caccgtgccc agcacctgaa ctcctggggg 3901 gaccgtcagt cttcctcttc cccccaaaac ccaaggacac cctcatgatc tcccggaccc 3961 ctgaggtcac atgcgtggtg gtggacgtga gccacgaaga ccctgaggtc aagttcaact 4021 ggtacgtgga cggcgtggag gtgcataatg ccaagacaaa gccgcgggag gagcagtaca 4081 acagcacgta ccgtgtggtc agcgtcctca ccgtcctgca ccaggactgg ctgaatggca 4141 aggagtacaa gtgcaaggtc tccaacaaag ccctcccagc ccccatcgag aaaaccatct 4201 ccaaagccaa agggcagccc cgagaaccac aggtgtacac cctgccccca tcccgggatg 4261 agctgaccaa gaaccaggtc agcctgacct gcctggtcaa aggcttctat cccagcgaca 4321 tcgccgtgga gtgggagagc aatgggcagc cggagaacaa ctacaagacc acgcctcccg 4381 tgctggactc cgacggctcc ttcttcctct acagcaagct caccgtggac aagagcaggt 4441 ggcagcaggg gaacgtcttc tcatgctccg tgatgcatga ggctctgcac aaccactaca 4501 cgcagaagag cctctccctg tctccgggta aagaattcgc ATCAACAAGT TTGTACAAAA 4561 AAGCAGGCTc tgaaaacttg tactttcaag gcgaggagga ggacaagaag gaggacgtgg 4621 gcacggtggt cggcatcgac ctggggacca cctactcctg cgtcggcgtg ttcaagaacg 4681 gccgcgtgga gatcatcgcc aacgatcagg gcaaccgcat cacgccgtcc tatgtcgcct 4741 tcactcctga aggggaacgt ctgattggcg atgccgccaa gaaccagctc acctccaacc 4801 ccgagaacac ggtctttgac gccaagcggc tcatcggccg cacgtggaat gacccgtctg 4861 tgcagcagga catcaagttc ttgccgttca aggtggttga aaagaaaact aaaccataca 4921 ttcaagttga tattggaggt gggcaaacaa agacatttgc tcctgaagaa atttctgcca 4981 tggttctcac taaaatgaaa gaaaccgctg aggcttattt gggaaagaag gttacccatg 5041 cagttgttac tgtaccagcc tattttaatg atgcccaacg ccaagcaacc aaagacgctg 5101 gaactattgc tggcctaaat gttatgagga tcatcaacga gcctacggca gctgctattg 5161 cttatggcct ggataagagg gagggggaga agaacatcct ggtgtttgac ctgggtggcg 5221 gaaccttcga tgtgtctctt ctcaccattg acaatggtgt cttcgaagtt gtggccacta 5281 atggagatac tcatctgggt ggagaagact ttgaccagcg tgtcatggaa cacttcatca 5341 aactgtacaa aaagaagacg ggcaaagatg tcaggaaaga caatagagct gtgcagaaac 5401 tccggcgcga ggtagaaaag gccaaacggg ccctgtcttc tcagcatcaa gcaagaattg 5461 aaattgagtc cttctatgaa ggagaagact tttctgagac cctgactcgg gccaaatttg 5521 aagagctcaa catggatctg ttccggtcta ctatgaagcc cgtccagaaa gtgttggaag 5581 attctgattt gaagaagtct gatattgatg aaattgttct tgttggtggc tcgactcgaa 5641 ttccaaagat tcagcaactg gttaaagagt tcttcaatgg caaggaacca tcccgtggca 5701 taaacccaga tgaagctgta gcgtatggtg ctgctgtcca ggctggtgtg ctctctggtg 5761 atcaagatac aggtgacctg gtactgcttg atgtatgtcc ccttacactt ggtattgaaa 5821 ctgtgggagg tgtcatgacc aaactgattc caaggaacac agtggtgcct accaagaagt 5881 ctcagatctt ttctacagct tctgataatc aaccaactgt tacaatcaag gtctatgaag 5941 gtgaaagacc cctgacaaaa gacaatcatc ttctgggtac atttgatctg actggaattc 6001 ctcctgctcc tcgtggggtc ccacagattg aagtcacctt tgagatagat gtgaatggta 6061 ttcttcgagt gacagctgaa gacaagggta cagggaacaa aaataagatc acaatcacca 6121 atgaccagaa tcgcctgaca cctgaagaaa tcgaaaggat ggttaatgat gctgagaagt 6181 ttgctgagga agacaaaaag ctcaaggagc gcattgatac tagaaatgag ttggaaagct 6241 atgcctattc tctaaagaat cagattggag ataaagaaaa gctgggaggt aaactttcct 6301 ctgaagataa ggagaccatg gaaaaagctg tagaagaaaa gattgaatgg ctggaaagcc 6361 accaagatgc tgacattgaa gacttcaaag ctaagaagaa ggaactggaa gaaattgttc 6421 aaccaattat cagcaaactc tatggaagtg caggccctcc cccaactggt gaagaggata 6481 cagcagaaaa agatgagttg TAGGACCCAG CTTTCTTGTA CAAAGTGGTT GATAGCTTGG 6541 ATCCAATCAA CCTCTGGATT ACAAAATTTG TGAAAGATTG ACTGGTATTC TTAACTATGT 6601 TGCTCCTTTT ACGCTATGTG GATACGCTGC TTTAATGCCT TTGTATCATG CTATTGCTTC 6661 CCGTATGGCT TTCATTTTCT CCTCCTTGTA TAAATCCTGG TTGCTGTCTC TTTATGAGGA 6721 GTTGTGGCCC GTTGTCAGGC AACGTGGCGT GGTGTGCACT GTGTTTGCTG ACGCAACCCC 6781 CACTGGTTGG GGCATTGCCA CCACCTGTCA GCTCCTTTCC GGGACTTTCG CTTTCCCCCT 6841 CCCTATTGCC ACGGCGGAAC TCATCGCCGC CTGCCTTGCC CGCTGCTGGA CAGGGGCTCG 6901 GCTGTTGGGC ACTGACAATT CCGTGGTGTT GTCGGGGAAG CTGACGTCCT TTCCATGGCT 6961 GCTCGCCTGT GTTGCCACCT GGATTCTGCG CGGGACGTCC TTCTGCTACG TCCCTTCGGC 7021 CCTCAATCCA GCGGACCTTC CTTCCCGCGG CCTGCTGCCG GCTCTGCGGC CTCTTCCGCG 7081 TCTTCGAGAT CTGCCTCGAC TGTGCCTTCT AGTTGCCAGC CATCTGTTGT TTGCCCCTCC 7141 CCCGTGCCTT CCTTGACCCT GGAAGGTGCC ACTCCCACTG TCCTTTCCTA ATAAAATGAG 7201 GAAATTGCAT CGCATTGTCT GAGTAGGTGT CATTCTATTC TGGGGGGTGG GGTGGGGCAG 7261 GACAGCAAGG GGGAGGATTG GGAAGACAAT AGCAGGCATG CTGGGGACTC GAGTTAAGGG 7321 CGAATTGCCG ATTAGGATCT TCCTAGAGCA TGGCTACGTA GATAAGTAGC ATGGCGGGTT 7381 AATCATTAAC TACAAGGAAC CCCTAGTGAT GGAGTTGGCC ACTCCCTCTC TGCGCGCTCG 7441 CTCGCTCACT GAGGCCGGGC GACCAAAGGT CGCCCGACGC CCGGGCTTTG CCCGGGCGGC 7501 CTCAGTGAGC GAGCGAGCGC GCAGCCTTAA TTAAGAGCTC ATGGCGCGCC TAGGCCTTGA 7561 CGGCCTTCCG CCAATTCGCC CTATAGTGAG TCGTATTACG TCGCGCTCAC TGGCCGTCGT 7621 TTTACAACGT CGTGACTGGG AAAACCCTGG CGTTACCCAA CTTAATCGCC TTGCAGCACA 7681 TCCCCCTTTC GCCAGCTGGC GTAATAGCGA AGAGGCCCGC ACCGAAACGC CCTTCCCAAC 7741 AGTTGCGCAG CCTGAATGGC GAATGGGAGC GCCCTGTAGC GGCCACTCAA CCCTATCTCG 7801 GTCTATTCTT TTGATTTATA AGGGATTTTG CCGATTTCGG CCTATTGGTT AAAAAATGAG 7861 CTGATTTAAC AAAAATTTAA CGCGAATTTT AACAAAATAT TAACGCTTAC AATTTAG //