LOCUS V019227 6488 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V019227 VERSION V019227 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..1168 /label="Il15(NM_008357)" /note="Il15(NM_008357)" /gene="Il15" CDS 1175..1240 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 1264..1852 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 1918..1966 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 2168..2596 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2610..2939 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3006..3800 /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 3976..4097 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4146..4162) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4170..4186 /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(4194..4224) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4239..4260 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4548..5136) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5307..6167) /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(6168..6272) /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 ccatgaaaat tttgaaacca tatatgagga atacatccat 721 ctcgtgctac ttgtgtttcc ttctaaacag tcacttttta actgaggctg gcattcatgt 781 cttcattttg ggctgtgtca gtgtaggtct ccctaaaaca gaggccaact ggatagatgt 841 aagatatgac ctggagaaaa ttgaaagcct tattcaatct attcatattg acaccacttt 901 atacactgac agtgactttc atcccagttg caaagttact gcaatgaact gctttctcct 961 ggaattgcag gttattttac atgagtacag taacatgact cttaatgaaa cagtaagaaa 1021 cgtgctctac cttgcaaaca gcactctgtc ttctaacaag aatgtagcag aatctggctg 1081 caaggaatgt gaggagctgg aggagaaaac cttcacagag tttttgcaaa gctttatacg 1141 cattgtccaa atgttcatca acacgtccct cgaggactac aaagaccatg acggtgatta 1201 taaagatcat gacatcgact acaaggatga cgatgacaag tagtgagggc ccgatatctc 1261 gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt 1321 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc 1381 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag 1441 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc 1501 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc 1561 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg 1621 ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt tccatggctg 1681 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc 1741 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt 1801 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcctggaaac 1861 gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg 1921 caataaaaag acagaataaa acgcacgggt gttgggtcgt ttgttcataa acgcggggtt 1981 cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc 2041 cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca 2101 gccaacgtcg gggcggcagg ccctgccata gcagatctgc gcagctgggg ctctaggggg 2161 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc 2221 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt 2281 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc ggggcatccc tttagggttc 2341 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt 2401 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt 2461 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt 2521 gatttataag ggattttggg gatttcggcc tattggttaa aaaatgagct gatttaacaa 2581 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag 2641 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg 2701 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag 2761 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc 2821 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg 2881 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa 2941 agctcccggg agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt 3001 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc 3061 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc 3121 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg 3181 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag 3241 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg 3301 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg 3361 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac 3421 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg 3481 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc 3541 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg 3601 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc 3661 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc 3721 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc 3781 ttcttgacga gttcttctga gcgggactct ggggttcgcg aaatgaccga ccaagcgacg 3841 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc 3901 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag 3961 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc 4021 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa 4081 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa 4141 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata 4201 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta 4261 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa 4321 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg 4381 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag 4441 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa 4501 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc 4561 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca 4621 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg 4681 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct 4741 caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt 4801 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag 4861 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc 4921 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac 4981 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga 5041 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc 5101 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg 5161 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca 5221 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt 5281 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca 5341 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg 5401 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca 5461 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt 5521 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt 5581 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca 5641 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca 5701 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga 5761 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact 5821 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga 5881 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg 5941 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc 6001 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga 6061 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat 6121 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt 6181 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt 6241 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac 6301 gtcgacggat cgggagatct cccgatcccc tatggtcgac tctcagtaca atctgctctg 6361 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt 6421 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc 6481 tgcttagg //