LOCUS V019215 6208 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V019215 VERSION V019215 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 652..669 /codon_start=1 /product="6xHis affinity tag" /transl_table=1 /translation="HHHHHH" /label="6xHis" CDS 697..951 /label="UFM1(NM_016617)" /note="UFM1(NM_016617)" /gene="UFM1" misc_feature 984..1572 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 1638..1686 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 1888..2316 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2330..2659 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 2726..3520 /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 3696..3817 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(3866..3882) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 3890..3906 /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(3914..3944) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 3959..3980 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4268..4856) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5027..5887) /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(5888..5992) /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 gcgccaccat gcatcatcac 661 catcaccatt taagcttggt accgagctcg gatcctatgt cgaaggtttc ctttaagatc 721 acgctgacgt cggacccacg gctgccgtac aaagtactca gtgttcctga aagtacacct 781 ttcacagcag tcttaaagtt tgcagcagaa gaatttaaag ttcctgctgc aacaagtgca 841 attattacca atgatggaat aggaataaat cctgcacaga ctgctggaaa tgtttttcta 901 aaacatggtt cagaactgcg gattattcct agagatcgtg ttggaagttg ttaactcgag 961 tctagagggc ccgatatctc gacaatcaac ctctggatta caaaatttgt gaaagattga 1021 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt 1081 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt 1141 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg 1201 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg 1261 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc 1321 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaagc 1381 tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct 1441 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg 1501 ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg 1561 ccgcctcccc gcctggaaac gggggaggct aactgaaaca cggaaggaga caataccgga 1621 aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacgggt gttgggtcgt 1681 ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac 1741 cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg 1801 gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata gcagatctgc 1861 gcagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg 1921 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt 1981 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc 2041 ggggcatccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg 2101 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga 2161 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc 2221 ctatctcggt ctattctttt gatttataag ggattttggg gatttcggcc tattggttaa 2281 aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt 2341 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa 2401 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag 2461 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct 2521 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc 2581 agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg 2641 aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca 2701 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc 2761 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc 2821 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga 2881 cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac 2941 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct 3001 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa 3061 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc 3121 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct 3181 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc 3241 caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg 3301 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct 3361 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct 3421 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca 3481 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgcg 3541 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct 3601 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc 3661 gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg 3721 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt 3781 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct 3841 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc 3901 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat 3961 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc 4021 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg 4081 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag 4141 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag 4201 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc 4261 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc 4321 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc 4381 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt 4441 cgggaagcgt ggcgctttct caatgctcac gctgtaggta tctcagttcg gtgtaggtcg 4501 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat 4561 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag 4621 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt 4681 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc 4741 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta 4801 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag 4861 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga 4921 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa 4981 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa 5041 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc 5101 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga 5161 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa 5221 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt 5281 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg 5341 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc 5401 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg 5461 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag 5521 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt 5581 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt 5641 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac 5701 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac 5761 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag 5821 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa 5881 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga 5941 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc 6001 cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtcgac 6061 tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt 6121 gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac 6181 cgacaattgc atgaagaatc tgcttagg //