LOCUS V019184 6551 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V019184 VERSION V019184 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..1231 /label="FTH1(NM_002032)" /note="FTH1(NM_002032)" /gene="FTH1" CDS 1238..1303 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 1327..1915 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 1981..2029 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 2231..2659 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2673..3002 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3069..3863 /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 4039..4160 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4209..4225) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4233..4249 /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(4257..4287) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4302..4323 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4611..5199) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5370..6230) /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(6231..6335) /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 ccatgacgac cgcgtccacc tcgcaggtgc gccagaacta 721 ccaccaggac tcagaggccg ccatcaaccg ccagatcaac ctggagctct acgcctccta 781 cgtttacctg tccatgtctt actactttga ccgcgatgat gtggctttga agaactttgc 841 caaatacttt cttcaccaat ctcatgagga gagggaacat gctgagaaac tgatgaagct 901 gcagaaccaa cgaggtggcc gaatcttcct tcaggatatc aagaaaccag actgtgatga 961 ctgggagagc gggctgaatg caatggagtg tgcattacat ttggaaaaaa atgtgaatca 1021 gtcactactg gaactgcaca aactggccac tgacaaaaat gacccccatt tgtgtgactt 1081 cattgagaca cattacctga atgagcaggt gaaagccatc aaagaattgg gtgaccacgt 1141 gaccaacttg cgcaagatgg gagcgcccga atctggcttg gcggaatatc tctttgacaa 1201 gcacaccctg ggagacagtg ataatgaaag cctcgaggac tacaaagacc atgacggtga 1261 ttataaagat catgacatcg actacaagga tgacgatgac aagtagtgag ggcccgatat 1321 ctcgacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat 1381 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct 1441 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag 1501 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc 1561 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc 1621 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct 1681 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg 1741 ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg 1801 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg 1861 cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcctgga 1921 aacgggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga 1981 cggcaataaa aagacagaat aaaacgcacg ggtgttgggt cgtttgttca taaacgcggg 2041 gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac 2101 gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc 2161 gcagccaacg tcggggcggc aggccctgcc atagcagatc tgcgcagctg gggctctagg 2221 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc 2281 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc 2341 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcggggcat ccctttaggg 2401 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca 2461 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc 2521 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct 2581 tttgatttat aagggatttt ggggatttcg gcctattggt taaaaaatga gctgatttaa 2641 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc 2701 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt 2761 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt 2821 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg 2881 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct 2941 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca 3001 aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat 3061 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga 3121 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc 3181 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga 3241 atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg 3301 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc 3361 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg 3421 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga 3481 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc 3541 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca 3601 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg 3661 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct 3721 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg 3781 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc 3841 gccttcttga cgagttcttc tgagcgggac tctggggttc gcgaaatgac cgaccaagcg 3901 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc 3961 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg 4021 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat 4081 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc 4141 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg 4201 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac 4261 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca 4321 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat 4381 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc 4441 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca 4501 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca 4561 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg 4621 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg 4681 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt 4741 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt 4801 tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc 4861 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt 4921 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt 4981 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc 5041 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa 5101 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt 5161 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct 5221 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta 5281 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa 5341 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc 5401 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact 5461 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc 5521 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt 5581 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta 5641 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg 5701 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt 5761 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc 5821 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt 5881 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc 5941 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc 6001 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa 6061 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac 6121 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa 6181 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt 6241 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa 6301 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct 6361 gacgtcgacg gatcgggaga tctcccgatc ccctatggtc gactctcagt acaatctgct 6421 ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt 6481 agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga 6541 atctgcttag g //