LOCUS V040235 6291 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V040235 VERSION V040235 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..1017 /label="DBI(NM_020548)" /note="DBI(NM_020548)" /gene="DBI" CDS 1024..1047 /codon_start=1 /product="8xHis affinity tag" /transl_table=1 /translation="HHHHHHHH" /label="8xHis" misc_feature 1067..1655 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 1721..1769 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 1971..2399 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2413..2742 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 2809..3603 /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 3779..3900 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(3949..3965) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 3973..3989 /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(3997..4027) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4042..4063 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4351..4939) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5110..5970) /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(5971..6075) /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 ccaccatgtg gggcgacctc 721 tggctcctcc cgcctgcctc tgccaatccg ggcactggga cagaggctga gtttgagaaa 781 gctgcagagg aggttaggca ccttaagacc aagccatcgg atgaggagat gctgttcatc 841 tatggccact acaaacaagc aactgtgggc gacataaata cagaacggcc cgggatgttg 901 gacttcacgg gcaaggccaa gtgggatgcc tggaatgagc tgaaagggac ttccaaggaa 961 gatgccatga aagcttacat caacaaagta gaagagctaa agaaaaaata cgggatactc 1021 gagcaccatc accatcacca tcaccattag ggcccgatat ctcgacaatc aacctctgga 1081 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg 1141 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt 1201 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag 1261 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc 1321 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga 1381 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa 1441 ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac 1501 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct 1561 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca 1621 gacgagtcgg atctcccttt gggccgcctc cccgcctgga aacgggggag gctaactgaa 1681 acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat 1741 aaaacgcacg ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca 1801 ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc 1861 cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc 1921 aggccctgcc atagcagatc tgcgcagctg gggctctagg gggtatcccc acgcgccctg 1981 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc 2041 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg 2101 ctttccccgt caagctctaa atcggggcat ccctttaggg ttccgattta gtgctttacg 2161 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg 2221 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt 2281 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt 2341 ggggatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta 2401 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga 2461 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc 2521 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc 2581 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc 2641 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag 2701 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt 2761 atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa 2821 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg 2881 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc 2941 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca 3001 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc 3061 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca 3121 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat 3181 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca 3241 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg 3301 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc 3361 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct 3421 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct 3481 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac 3541 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc 3601 tgagcgggac tctggggttc gcgaaatgac cgaccaagcg acgcccaacc tgccatcacg 3661 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga 3721 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa 3781 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa 3841 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta 3901 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt 3961 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa 4021 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact 4081 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc 4141 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg 4201 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc 4261 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag 4321 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca 4381 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca 4441 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg 4501 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag 4561 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt 4621 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca 4681 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg 4741 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt 4801 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc 4861 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg 4921 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg 4981 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta 5041 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg 5101 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg 5161 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc 5221 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc 5281 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc 5341 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag 5401 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat 5461 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg 5521 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt 5581 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag 5641 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg 5701 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt 5761 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct 5821 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac 5881 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat 5941 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat 6001 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca 6061 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga 6121 tctcccgatc ccctatggtc gactctcagt acaatctgct ctgatgccgc atagttaagc 6181 cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa 6241 gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag g //