LOCUS V020513 7388 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V020513 VERSION V020513 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..2068 /label="ERRFI1(NM_018948)" /note="ERRFI1(NM_018948)" /gene="ERRFI1" CDS 2075..2140 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 2164..2752 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 2818..2866 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 3068..3496 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3510..3839 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3906..4700 /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 4876..4997 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5046..5062) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5070..5086 /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(5094..5124) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5139..5160 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5448..6036) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6207..7067) /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(7068..7172) /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 ccatgtcaat agcaggagtt gctgctcagg agatcagagt 721 cccattaaaa actggatttc tacataatgg ccgagccatg gggaatatga ggaagaccta 781 ctggagcagt cgcagtgagt ttaaaaacaa ctttttaaat attgacccga taaccatggc 841 ctacagtctg aactcttctg ctcaggagcg cctaatacca cttgggcatg cttccaaatc 901 tgctccgatg aatggccact gctttgcaga aaatggtcca tctcaaaagt ccagcttgcc 961 ccctcttctt attcccccaa gtgaaaactt gggaccacat gaagaggatc aagttgtatg 1021 tggttttaag aaactcacag tgaatggggt ttgtgcttcc acccctccac tgacacccat 1081 aaaaaactcc ccttcccttt tcccctgtgc ccctctttgt gaacggggtt ctaggcctct 1141 tccaccgttg ccaatctctg aagccctctc tctggatgac acagactgtg aggtggaatt 1201 cctaactagc tcagatacag acttcctttt agaagactct acactttctg atttcaaata 1261 tgatgttcct ggcaggcgaa gcttccgtgg gtgtggacaa atcaactatg catattttga 1321 taccccagct gtttctgcag cagatctcag ctatgtgtct gaccaaaatg gaggtgtccc 1381 agatccaaat cctcctccac ctcagaccca ccgaagatta agaaggtctc attcgggacc 1441 agctggctcc tttaacaagc cagccataag gatatccaac tgttgtatac acagagcttc 1501 tcctaactcc gatgaagaca aacctgaggt tccccccaga gttcccatac ctcctagacc 1561 agtaaagcca gattatagaa gatggtcagc agaagttact tcgagcacct atagtgatga 1621 agacaggcct cccaaagtac cgccaagaga acctttgtca ccgagtaact cgcgcacacc 1681 gagtcccaaa agccttccgt cttacctcaa tggggtcatg cccccgacac agagctttgc 1741 ccctgatccc aagtatgtca gcagcaaagc actgcaaaga cagaacagcg aaggatctgc 1801 cagtaaggtt ccttgcattc tgcccattat tgaaaatggg aagaaggtta gttcaacaca 1861 ttattaccta ctacctgaac gaccaccata cctggacaaa tatgaaaaat tttttaggga 1921 agcagaagaa acaaatggag gcgcccaaat ccagccatta cctgctgact gcggtatatc 1981 ttcagccaca gaaaagccag actcaaaaac aaaaatggat ctgggtggcc acgtgaagcg 2041 taaacattta tcctatgtgg tttctcctct cgaggactac aaagaccatg acggtgatta 2101 taaagatcat gacatcgact acaaggatga cgatgacaag tagtgagggc ccgatatctc 2161 gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt 2221 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc 2281 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag 2341 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc 2401 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc 2461 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg 2521 ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt tccatggctg 2581 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc 2641 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt 2701 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcctggaaac 2761 gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg 2821 caataaaaag acagaataaa acgcacgggt gttgggtcgt ttgttcataa acgcggggtt 2881 cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc 2941 cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca 3001 gccaacgtcg gggcggcagg ccctgccata gcagatctgc gcagctgggg ctctaggggg 3061 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc 3121 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt 3181 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc ggggcatccc tttagggttc 3241 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt 3301 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt 3361 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt 3421 gatttataag ggattttggg gatttcggcc tattggttaa aaaatgagct gatttaacaa 3481 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag 3541 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg 3601 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag 3661 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc 3721 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg 3781 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa 3841 agctcccggg agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt 3901 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc 3961 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc 4021 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg 4081 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag 4141 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg 4201 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg 4261 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac 4321 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg 4381 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc 4441 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg 4501 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc 4561 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc 4621 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc 4681 ttcttgacga gttcttctga gcgggactct ggggttcgcg aaatgaccga ccaagcgacg 4741 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc 4801 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag 4861 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc 4921 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa 4981 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa 5041 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata 5101 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta 5161 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa 5221 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg 5281 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag 5341 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa 5401 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc 5461 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca 5521 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg 5581 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct 5641 caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt 5701 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag 5761 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc 5821 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac 5881 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga 5941 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc 6001 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg 6061 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca 6121 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt 6181 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca 6241 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg 6301 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca 6361 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt 6421 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt 6481 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca 6541 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca 6601 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga 6661 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact 6721 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga 6781 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg 6841 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc 6901 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga 6961 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat 7021 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt 7081 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt 7141 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac 7201 gtcgacggat cgggagatct cccgatcccc tatggtcgac tctcagtaca atctgctctg 7261 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt 7321 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc 7381 tgcttagg //