LOCUS V043934 6828 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V043934 VERSION V043934 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers enhancer 50..429 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 430..633 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" CDS 706..1554 /label="VDAC1(NM_003374)" /note="VDAC1(NM_003374)" /gene="VDAC1" CDS 1561..1584 /codon_start=1 /product="8xHis affinity tag" /transl_table=1 /translation="HHHHHHHH" /label="8xHis" misc_feature 1604..2192 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 2258..2306 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 2508..2936 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2950..3279 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3346..4140 /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 4316..4437 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4486..4502) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4510..4526 /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(4534..4564) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4579..4600 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4888..5476) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5647..6507) /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(6508..6612) /gene="bla" /label="AmpR promoter" ORIGIN 1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta 61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag 121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc 181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga 241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat 301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc 361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct 421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca 481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat 541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg 601 cgtgtacggt gggaggtcta tataagcaga gctctggcta gcgtttaaac ttaagcttgg 661 taccgagctc ggatccacta gtccagtgtg gtggaattcg ccaccatggc tgtgccaccc 721 acgtatgccg atcttggcaa atctgccagg gatgtcttca ccaagggcta tggatttggc 781 ttaataaagc ttgatttgaa aacaaaatct gagaatggat tggaatttac aagctcaggc 841 tcagccaaca ctgagaccac caaagtgacg ggcagtctgg aaaccaagta cagatggact 901 gagtacggcc tgacgtttac agagaaatgg aataccgaca atacactagg caccgagatt 961 actgtggaag atcagcttgc acgtggactg aagctgacct tcgattcatc cttctcacct 1021 aacactggga aaaaaaatgc taaaatcaag acagggtaca agcgggagca cattaacctg 1081 ggctgcgaca tggatttcga cattgctggg ccttccatcc ggggtgctct ggtgctaggt 1141 tacgagggct ggctggccgg ctaccagatg aattttgaga ctgcaaaatc ccgagtgacc 1201 cagagcaact ttgcagttgg ctacaagact gatgaattcc agcttcacac taatgtgaat 1261 gacgggacag agtttggcgg ctccatttac cagaaagtga acaagaagtt ggagaccgct 1321 gtcaatcttg cctggacagc aggaaacagt aacacgcgct tcggaatagc agccaagtat 1381 cagattgacc ctgacgcctg cttctcggct aaagtgaaca actccagcct gataggttta 1441 ggatacactc agactctaaa gccaggtatt aaactgacac tgtcagctct tctggatggc 1501 aagaacgtca atgctggtgg ccacaagctt ggtctaggac tggaatttca agcactcgag 1561 caccatcacc atcaccatca ccattagggc ccgatatctc gacaatcaac ctctggatta 1621 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg 1681 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc 1741 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca 1801 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac 1861 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact 1921 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc 1981 cgtggtgttg tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg 2041 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc 2101 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac 2161 gagtcggatc tccctttggg ccgcctcccc gcctggaaac gggggaggct aactgaaaca 2221 cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa 2281 acgcacgggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc 2341 tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc 2401 accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg 2461 ccctgccata gcagatctgc gcagctgggg ctctaggggg tatccccacg cgccctgtag 2521 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag 2581 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt 2641 tccccgtcaa gctctaaatc ggggcatccc tttagggttc cgatttagtg ctttacggca 2701 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata 2761 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca 2821 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttggg 2881 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt 2941 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt 3001 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca 3061 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta 3121 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga 3181 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag 3241 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata 3301 tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat 3361 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca 3421 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg 3481 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg 3541 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact 3601 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct 3661 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg 3721 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt 3781 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc 3841 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc 3901 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga 3961 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc 4021 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt 4081 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga 4141 gcgggactct ggggttcgcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga 4201 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc 4261 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt 4321 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa 4381 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca 4441 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc 4501 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg 4561 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc 4621 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg 4681 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc 4741 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac 4801 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa 4861 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca 4921 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc 4981 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata 5041 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct caatgctcac gctgtaggta 5101 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca 5161 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga 5221 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg 5281 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg 5341 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg 5401 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag 5461 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa 5521 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat 5581 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc 5641 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc 5701 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc 5761 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc 5821 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc 5881 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt 5941 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc 6001 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa 6061 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt 6121 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg 6181 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc 6241 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa 6301 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt 6361 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt 6421 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag 6481 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta 6541 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat 6601 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct 6661 cccgatcccc tatggtcgac tctcagtaca atctgctctg atgccgcata gttaagccag 6721 tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct 6781 acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagg //