LOCUS V028605 6142 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V028605 VERSION V028605 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 931..1635 /label="TFPI2(NM_006528)" /note="TFPI2(NM_006528)" /gene="TFPI2" CDS 1645..1662 /codon_start=1 /product="6xHis affinity tag" /transl_table=1 /translation="HHHHHH" /label="6xHis" polyA_signal 1709..1933 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 1979..2407 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2421..2750 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 2817..3611 /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 3785..3906 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(3955..3971) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 3979..3995 /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(4003..4033) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4048..4069 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4357..4942) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5113..5973) /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(5974..6078) /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 cactagtcca gtgtggtgga attcgccacc atggaccccg ctcgccccct ggggctgtcg 961 attctgctgc ttttcctgac ggaggctgca ctgggcgatg ctgctcagga gccaacagga 1021 aataacgcgg agatctgtct cctgccccta gactacggac cctgccgggc cctacttctc 1081 cgttactact acgacaggta cacgcagagc tgccgccagt tcctgtacgg gggctgcgag 1141 ggcaacgcca acaatttcta cacctgggag gcttgcgacg atgcttgctg gaggatagaa 1201 aaagttccca aagtttgccg gctgcaagtg agtgtggacg accagtgtga ggggtccaca 1261 gaaaagtatt tctttaatct aagttccatg acatgtgaaa aattcttttc cggtgggtgt 1321 caccggaacc ggattgagaa caggtttcca gatgaagcta cttgtatggg cttctgcgca 1381 ccaaagaaaa ttccatcatt ttgctacagt ccaaaagatg agggactgtg ctctgccaat 1441 gtgactcgct attattttaa tccaagatac agaacctgtg atgctttcac ctatactggc 1501 tgtggaggga atgacaataa ctttgttagc agggaggatt gcaaacgtgc atgtgcaaaa 1561 gctttgaaaa agaaaaagaa gatgccaaag cttcgctttg ccagtagaat ccggaaaatt 1621 cggaagaagc aatttgcggc cgcacatcac catcatcacc attgactcga gtctagaggg 1681 cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt 1741 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa 1801 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg 1861 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg 1921 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac 1981 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct 2041 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg 2101 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt 2161 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca 2221 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga 2281 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa 2341 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac 2401 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag 2461 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc 2521 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag 2581 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc 2641 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc 2701 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg 2761 gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga 2821 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct 2881 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc 2941 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg 3001 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg 3061 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc 3121 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc 3181 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg 3241 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc 3301 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg 3361 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc 3421 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag 3481 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg 3541 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg 3601 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc 3661 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt 3721 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca 3781 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt 3841 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt 3901 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata 3961 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag 4021 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg 4081 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca 4141 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc 4201 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg 4261 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa 4321 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga 4381 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag 4441 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct 4501 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg 4561 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc 4621 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt 4681 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta 4741 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac 4801 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc 4861 ttgatccggc aaacaaacca ccgctggtag cggttttttt gtttgcaagc agcagattac 4921 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca 4981 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac 5041 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac 5101 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt 5161 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt 5221 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt 5281 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc 5341 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa 5401 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg 5461 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt 5521 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc 5581 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt 5641 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg 5701 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac 5761 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc 5821 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt 5881 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg 5941 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag 6001 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa 6061 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg 6121 agatctcccg atcccctatg gt //