LOCUS V022535 7148 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V022535 VERSION V022535 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 683..1828 /label="GJA1(NM_000165)" /note="GJA1(NM_000165)" /gene="GJA1" CDS 1835..1900 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 1924..2512 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 2578..2626 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 2828..3256 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3270..3599 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3666..4460 /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 4636..4757 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4806..4822) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4830..4846 /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(4854..4884) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4899..4920 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5208..5796) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5967..6827) /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(6828..6932) /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 ggatccgcca ccatgggtga ctggagcgcc ttaggcaaac tccttgacaa 721 ggttcaagcc tactcaactg ctggagggaa ggtgtggctg tcagtacttt tcattttccg 781 aatcctgctg ctggggacag cggttgagtc agcctgggga gatgagcagt ctgcctttcg 841 ttgtaacact cagcaacctg gttgtgaaaa tgtctgctat gacaagtctt tcccaatctc 901 tcatgtgcgc ttctgggtcc tgcagatcat atttgtgtct gtacccacac tcttgtacct 961 ggctcatgtg ttctatgtga tgcgaaagga agagaaactg aacaagaaag aggaagaact 1021 caaggttgcc caaactgatg gtgtcaatgt ggacatgcac ttgaagcaga ttgagataaa 1081 gaagttcaag tacggtattg aagagcatgg taaggtgaaa atgcgagggg ggttgctgcg 1141 aacctacatc atcagtatcc tcttcaagtc tatctttgag gtggccttct tgctgatcca 1201 gtggtacatc tatggattca gcttgagtgc tgtttacact tgcaaaagag atccctgccc 1261 acatcaggtg gactgtttcc tctctcgccc cacggagaaa accatcttca tcatcttcat 1321 gctggtggtg tccttggtgt ccctggcctt gaatatcatt gaactcttct atgttttctt 1381 caagggcgtt aaggatcggg ttaagggaaa gagcgaccct taccatgcga ccagtggtgc 1441 gctgagccct gccaaagact gtgggtctca aaaatatgct tatttcaatg gctgctcctc 1501 accaaccgct cccctctcgc ctatgtctcc tcctgggtac aagctggtta ctggcgacag 1561 aaacaattct tcttgccgca attacaacaa gcaagcaagt gagcaaaact gggctaatta 1621 cagtgcagaa caaaatcgaa tggggcaggc gggaagcacc atctctaact cccatgcaca 1681 gccttttgat ttccccgatg ataaccagaa ttctaaaaaa ctagctgctg gacatgaatt 1741 acagccacta gccattgtgg accagcgacc ttcaagcaga gccagcagtc gtgccagcag 1801 cagacctcgg cctgatgacc tggagatcct cgaggactac aaagaccatg acggtgatta 1861 taaagatcat gacatcgact acaaggatga cgatgacaag tagtgagggc ccgatatctc 1921 gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt 1981 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc 2041 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag 2101 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc 2161 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc 2221 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg 2281 ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt tccatggctg 2341 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc 2401 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt 2461 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcctggaaac 2521 gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg 2581 caataaaaag acagaataaa acgcacgggt gttgggtcgt ttgttcataa acgcggggtt 2641 cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc 2701 cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca 2761 gccaacgtcg gggcggcagg ccctgccata gcagatctgc gcagctgggg ctctaggggg 2821 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc 2881 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt 2941 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc ggggcatccc tttagggttc 3001 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt 3061 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt 3121 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt 3181 gatttataag ggattttggg gatttcggcc tattggttaa aaaatgagct gatttaacaa 3241 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag 3301 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg 3361 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag 3421 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc 3481 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg 3541 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa 3601 agctcccggg agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt 3661 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc 3721 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc 3781 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg 3841 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag 3901 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg 3961 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg 4021 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac 4081 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg 4141 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc 4201 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg 4261 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc 4321 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc 4381 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc 4441 ttcttgacga gttcttctga gcgggactct ggggttcgcg aaatgaccga ccaagcgacg 4501 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc 4561 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag 4621 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc 4681 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa 4741 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa 4801 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata 4861 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta 4921 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa 4981 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg 5041 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag 5101 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa 5161 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc 5221 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca 5281 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg 5341 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct 5401 caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt 5461 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag 5521 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc 5581 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac 5641 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga 5701 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc 5761 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg 5821 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca 5881 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt 5941 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca 6001 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg 6061 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca 6121 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt 6181 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt 6241 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca 6301 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca 6361 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga 6421 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact 6481 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga 6541 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg 6601 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc 6661 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga 6721 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat 6781 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt 6841 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt 6901 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac 6961 gtcgacggat cgggagatct cccgatcccc tatggtcgac tctcagtaca atctgctctg 7021 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt 7081 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc 7141 tgcttagg //