LOCUS V021191 7445 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V021191 VERSION V021191 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..2092 /label="Traf4(NM_009423)" /note="Traf4(NM_009423)" /gene="Traf4" CDS 2132..2197 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 2221..2809 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 2875..2923 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 3125..3553 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3567..3896 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3963..4757 /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 4933..5054 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5103..5119) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5127..5143 /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(5151..5181) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5196..5217 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5505..6093) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6264..7124) /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(7125..7229) /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 ccatgcccgg cttcgactac aagttcctgg agaagcccaa 721 gcggcggctg ctgtgcccac tgtgcgggaa gcccatgcgc gagcctgtgc aggtgtctac 781 ttgtggccac cgcttctgcg acacctgcct gcaggagttt ctcagtgaag gagtcttcaa 841 atgccctgag gaccaacttc ctctggacta tgccaagatc tacccagacc cagagctgga 901 ggtccaggtg ttaggcttgg ctatccgctg catccacagt gaggagggct gccgctggag 961 cgggccactt cgtcatctac agggccactt gaacacttgc agcttcaatg tagtcccctg 1021 ccctaatcgc tgcccagcca agctgagccg gcgtgacctg cctgcccacc tgcagcacga 1081 ctgccctaag cgccgcctca agtgtgaatt ctgtggctgt gacttcagtg gggaggccta 1141 cgagagtcac gagggcgtgt gcccccaaga gagtgtgtac tgtgagaaca agtgtggggc 1201 ccgcatgatg aggcgactgt tggcccagca tgccacgtct gagtgcccca agcgcaccca 1261 gccctgtgcc tactgtacca aggaatttgt ttatgacacc atccagagcc accagtacca 1321 gtgcccacgc ctgcctgttc cctgccccaa ccaatgtgga gtgggcactg tggcccggga 1381 agacctgccc acccatctga aggatagctg cagcactgcc tttgttctct gccctttcaa 1441 agagtctggc tgcaagcaca ggtgccctaa gctggcaatg ggacgtcacg tggaggagag 1501 cgtaaagcca catctggcta tgatgtgtgc cctggtgagc cgacagcggc aagagctgca 1561 ggagctgcga agagagctgg aagaactatc cataggcagc gatggagtgc tcatctggaa 1621 gattggcagc tatgggcgtc gtctacaaga ggccaaggcc aagcctaacc tggagtgctt 1681 cagcccagcc ttctatacgc ataagtacgg ctacaagctg caggtgtctg cattcctcaa 1741 cggcaacggc agtggtgagg gcacacacct ctccatctac atccgtgtct tgccgggcgc 1801 ctttgacaat ctccttgagt ggccctttgc ccgccgggtc accttctccc tgctggatca 1861 gagtgaccca ggtttggcta agccacagca tgtcactgag accttccacc ctgatccaaa 1921 ctggaagaac ttccagaagc ccggcacgtg gcgaggctcc ctggatgaga gttctctggg 1981 cttcggctac cccaagttca tttcccacca ggacatccga aagcggaact acgtgcggga 2041 tgatgcggtt ttcatccgtg cctctgtcga actgccccgg aagatcctca gcgaattctg 2101 cagatatcca gcacagtggc ggccgctcga ggactacaaa gaccatgacg gtgattataa 2161 agatcatgac atcgactaca aggatgacga tgacaagtag tgagggcccg atatctcgac 2221 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct 2281 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt 2341 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg 2401 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact 2461 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct 2521 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg 2581 ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc atggctgctc 2641 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc 2701 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt 2761 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc tggaaacggg 2821 ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa 2881 taaaaagaca gaataaaacg cacgggtgtt gggtcgtttg ttcataaacg cggggttcgg 2941 tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc 3001 gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc 3061 aacgtcgggg cggcaggccc tgccatagca gatctgcgca gctggggctc tagggggtat 3121 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg 3181 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc 3241 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg gcatcccttt agggttccga 3301 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt 3361 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat 3421 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat 3481 ttataaggga ttttggggat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa 3541 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct 3601 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa 3661 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa 3721 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt 3781 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct 3841 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc 3901 tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc 3961 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat 4021 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt 4081 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac 4141 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg 4201 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc 4261 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa 4321 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc 4381 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg 4441 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg 4501 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa 4561 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg 4621 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct 4681 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc 4741 ttgacgagtt cttctgagcg ggactctggg gttcgcgaaa tgaccgacca agcgacgccc 4801 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga 4861 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc 4921 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc 4981 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc 5041 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca 5101 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga 5161 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt 5221 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga 5281 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc 5341 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg 5401 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc 5461 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc 5521 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga 5581 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc 5641 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa 5701 tgctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg 5761 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc 5821 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga 5881 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact 5941 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt 6001 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag 6061 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg 6121 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa 6181 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata 6241 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg 6301 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata 6361 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg 6421 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct 6481 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt 6541 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc 6601 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga 6661 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt 6721 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc 6781 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa 6841 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca 6901 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca 6961 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct 7021 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc 7081 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa 7141 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt 7201 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc 7261 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg 7321 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 7381 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 7441 ttagg //