LOCUS V021455 6608 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V021455 VERSION V021455 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..1255 /label="Kcnmb1(NM_031169)" /note="Kcnmb1(NM_031169)" /gene="Kcnmb1" CDS 1295..1360 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 1384..1972 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 2038..2086 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 2288..2716 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2730..3059 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3126..3920 /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 4096..4217 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4266..4282) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4290..4306 /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(4314..4344) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4359..4380 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4668..5256) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5427..6287) /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(6288..6392) /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 ccatggggaa gaagctggtg atggcccaga agcgcggaga 721 gacacgagcc ctctgcctgg gagtggcaat ggtagtgtgt gctgccatca cctactacgt 781 cctgggtaca actgtgctgc ccctctacca gaaaagtgtg tggacccagg aatccatatg 841 tcacttgatt gaaactaata tcaaggacca ggaagagctg gagggcaaga aggtgcccca 901 gtacccatgc ctttgggtca atgtatcagc tgtgggcaga tgggccatgc tgtatcacac 961 ggaagacact cgggatcaaa accaacagtg ctcctatatc cccaggaacc tggacaacta 1021 ccagacagcc ttggcagatg tgaagaaggt cagagccaat ttctataagc accatgaatt 1081 ctattgcctt tctgcacctc aagtcaacga gaccagcgtc gtgtaccagc gcctctacgg 1141 gccccaagtc ctcctcttct ccttcttctg gcccaccttc ctgctgaccg ggggtcttat 1201 tcttattgcc atggtgaagc tcaacaggtc cctatccatc ttggcggctc agaaggaatt 1261 ctgcagatat ccagcacagt ggcggccgct cgaggactac aaagaccatg acggtgatta 1321 taaagatcat gacatcgact acaaggatga cgatgacaag tagtgagggc ccgatatctc 1381 gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt 1441 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc 1501 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag 1561 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc 1621 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc 1681 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg 1741 ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt tccatggctg 1801 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc 1861 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt 1921 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcctggaaac 1981 gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg 2041 caataaaaag acagaataaa acgcacgggt gttgggtcgt ttgttcataa acgcggggtt 2101 cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc 2161 cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca 2221 gccaacgtcg gggcggcagg ccctgccata gcagatctgc gcagctgggg ctctaggggg 2281 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc 2341 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt 2401 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc ggggcatccc tttagggttc 2461 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt 2521 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt 2581 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt 2641 gatttataag ggattttggg gatttcggcc tattggttaa aaaatgagct gatttaacaa 2701 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag 2761 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg 2821 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag 2881 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc 2941 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg 3001 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa 3061 agctcccggg agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt 3121 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc 3181 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc 3241 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg 3301 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag 3361 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg 3421 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg 3481 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac 3541 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg 3601 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc 3661 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg 3721 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc 3781 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc 3841 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc 3901 ttcttgacga gttcttctga gcgggactct ggggttcgcg aaatgaccga ccaagcgacg 3961 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc 4021 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag 4081 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc 4141 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa 4201 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa 4261 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata 4321 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta 4381 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa 4441 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg 4501 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag 4561 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa 4621 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc 4681 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca 4741 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg 4801 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct 4861 caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt 4921 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag 4981 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc 5041 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac 5101 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga 5161 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc 5221 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg 5281 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca 5341 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt 5401 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca 5461 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg 5521 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca 5581 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt 5641 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt 5701 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca 5761 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca 5821 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga 5881 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact 5941 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga 6001 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg 6061 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc 6121 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga 6181 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat 6241 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt 6301 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt 6361 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac 6421 gtcgacggat cgggagatct cccgatcccc tatggtcgac tctcagtaca atctgctctg 6481 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt 6541 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc 6601 tgcttagg //