LOCUS V021006 6308 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V021006 VERSION V021006 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers enhancer 235..614 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 615..818 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" promoter 863..881 /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" CDS 932..1780 /label="VDAC1(NM_003374)" /note="VDAC1(NM_003374)" /gene="VDAC1" CDS 1793..1822 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /transl_table=1 /translation="EQKLISEEDL" /label="Myc" CDS 1838..1855 /codon_start=1 /product="6xHis affinity tag" /transl_table=1 /translation="HHHHHH" /label="6xHis" polyA_signal 1905..2129 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2175..2603 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2617..2946 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3013..3807 /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 3981..4102 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4151..4167) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4175..4191 /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(4199..4229) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4244..4265 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4553..5141) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5312..6172) /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(6173..6277) /gene="bla" /label="AmpR promoter" ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg 61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt 481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca 841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc 901 gtttaaacgg gccctctaga ctcgagccac catggctgtg ccacccacgt atgccgatct 961 tggcaaatct gccagggatg tcttcaccaa gggctatgga tttggcttaa taaagcttga 1021 tttgaaaaca aaatctgaga atggattgga atttacaagc tcaggctcag ccaacactga 1081 gaccaccaaa gtgacgggca gtctggaaac caagtacaga tggactgagt acggcctgac 1141 gtttacagag aaatggaata ccgacaatac actaggcacc gagattactg tggaagatca 1201 gcttgcacgt ggactgaagc tgaccttcga ttcatccttc tcacctaaca ctgggaaaaa 1261 aaatgctaaa atcaagacag ggtacaagcg ggagcacatt aacctgggct gcgacatgga 1321 tttcgacatt gctgggcctt ccatccgggg tgctctggtg ctaggttacg agggctggct 1381 ggccggctac cagatgaatt ttgagactgc aaaatcccga gtgacccaga gcaactttgc 1441 agttggctac aagactgatg aattccagct tcacactaat gtgaatgacg ggacagagtt 1501 tggcggctcc atttaccaga aagtgaacaa gaagttggag accgctgtca atcttgcctg 1561 gacagcagga aacagtaaca cgcgcttcgg aatagcagcc aagtatcaga ttgaccctga 1621 cgcctgcttc tcggctaaag tgaacaactc cagcctgata ggtttaggat acactcagac 1681 tctaaagcca ggtattaaac tgacactgtc agctcttctg gatggcaaga acgtcaatgc 1741 tggtggccac aagcttggtc taggactgga atttcaagca aagcttgggc ccgaacaaaa 1801 actcatctca gaagaggatc tgaatagcgc cgtcgaccat catcatcatc atcattgagt 1861 ttaaacggtc tccagcttaa gtttaaaccg ctgatcagcc tcgactgtgc cttctagttg 1921 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc 1981 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc 2041 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag 2101 gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc 2161 tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac 2221 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc 2281 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt 2341 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg 2401 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac 2461 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta 2521 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat 2581 ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag 2641 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc 2701 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat 2761 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt 2821 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc 2881 gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt 2941 tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga 3001 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg 3061 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg 3121 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc 3181 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct 3241 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa 3301 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg 3361 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa 3421 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat 3481 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg 3541 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc 3601 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac 3661 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg 3721 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc 3781 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag 3841 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg 3901 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc 3961 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca 4021 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt 4081 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg 4141 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca 4201 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca 4261 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc 4321 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt 4381 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact 4441 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag 4501 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata 4561 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc 4621 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg 4681 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc 4741 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg 4801 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc 4861 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga 4921 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg 4981 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa 5041 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg 5101 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt 5161 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat 5221 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct 5281 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta 5341 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa 5401 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac 5461 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa 5521 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag 5581 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg 5641 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag 5701 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg 5761 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc 5821 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat 5881 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata 5941 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa 6001 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca 6061 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc 6121 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc 6181 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg 6241 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac 6301 ctgacgtc //