LOCUS V043960 6326 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V043960 VERSION V043960 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers enhancer 202..581 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 582..785 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" promoter 830..848 /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" CDS 908..1819 /label="TFPI(NM_006287)" /note="TFPI(NM_006287)" /gene="TFPI" CDS 1829..1846 /codon_start=1 /product="6xHis affinity tag" /transl_table=1 /translation="HHHHHH" /label="6xHis" polyA_signal 1893..2117 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2163..2591 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2605..2934 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3001..3795 /codon_start=1 /gene="aph(3')-II (or nptII)" /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin®)" /product="aminoglycoside phosphotransferase from Tn5" /transl_table=1 /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*" /label="NeoR/KanR" polyA_signal 3969..4090 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4139..4155) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4163..4179 /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG)." /label="lac operator" promoter complement(4187..4217) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4232..4253 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4541..5126) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5297..6157) /codon_start=1 /gene="bla" /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /product="β-lactamase" /transl_table=1 /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA SLIKHW*" /label="AmpR" promoter complement(6158..6262) /gene="bla" /label="AmpR promoter" ORIGIN 1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt 61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct 121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg 181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta 241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg 301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc 361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa 421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca 481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta 541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt 601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg 661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca 721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca 781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc 841 actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg agctcggatc 901 cgccaccatg atttacacaa tgaagaaagt acatgcactt tgggcttctg tatgcctgct 961 gcttaatctt gcccctgccc ctcttaatgc tgattctgag gaagatgaag aacacacaat 1021 tatcacagat acggagttgc caccactgaa acttatgcat tcattttgtg cattcaaggc 1081 ggatgatggc ccatgtaaag caatcatgaa aagatttttc ttcaatattt tcactcgaca 1141 gtgcgaagaa tttatatatg ggggatgtga aggaaatcag aatcgatttg aaagtctgga 1201 agagtgcaaa aaaatgtgta caagagataa tgcaaacagg attataaaga caacattgca 1261 acaagaaaag ccagatttct gctttttgga agaagatcct ggaatatgtc gaggttatat 1321 taccaggtat ttttataaca atcagacaaa acagtgtgaa cgtttcaagt atggtggatg 1381 cctgggcaat atgaacaatt ttgagacact ggaagaatgc aagaacattt gtgaagatgg 1441 tccgaatggt ttccaggtgg ataattatgg aacccagctc aatgctgtga ataactccct 1501 gactccgcaa tcaaccaagg ttcccagcct ttttgaattt cacggtccct catggtgtct 1561 cactccagca gacagaggat tgtgtcgtgc caatgagaac agattctact acaattcagt 1621 cattgggaaa tgccgcccat ttaagtacag tggatgtggg ggaaatgaaa acaattttac 1681 ttccaaacaa gaatgtctga gggcatgtaa aaaaggtttc atccaaagaa tatcaaaagg 1741 aggcctaatt aaaaccaaaa gaaaaagaaa gaagcagaga gtgaaaatag catatgaaga 1801 aatttttgtt aaaaatatgg cggccgcaca tcaccatcat caccattgac tcgagtctag 1861 agggcccgtt taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt 1921 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc 1981 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg 2041 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga 2101 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc 2161 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac 2221 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc 2281 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt 2341 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg 2401 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag 2461 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt 2521 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt 2581 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc 2641 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag 2701 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc 2761 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct 2821 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct 2881 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc 2941 ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc 3001 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc 3061 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca 3121 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg 3181 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg 3241 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag 3301 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg 3361 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc 3421 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa 3481 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac 3541 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat 3601 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac 3661 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc 3721 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt 3781 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc 3841 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg 3901 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg 3961 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa 4021 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca 4081 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt 4141 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg 4201 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt 4261 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg 4321 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg 4381 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa 4441 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc 4501 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc 4561 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat 4621 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc 4681 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct 4741 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg 4801 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc 4861 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga 4921 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa 4981 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta 5041 gctcttgatc cggcaaacaa accaccgctg gtagcggttt ttttgtttgc aagcagcaga 5101 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg 5161 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct 5221 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt 5281 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc 5341 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg 5401 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag 5461 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt 5521 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag 5581 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt 5641 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca 5701 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg 5761 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat 5821 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta 5881 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca 5941 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct 6001 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat 6061 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa 6121 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt 6181 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa 6241 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat 6301 cgggagatct cccgatcccc tatggt //