LOCUS V018391 7505 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V018391 VERSION V018391 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 671..2185 /label="Pparg(NM_011146)" /note="Pparg(NM_011146)" /gene="Pparg" CDS 2192..2257 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" misc_feature 2281..2869 /note="woodchuck hepatitis virus posttranscriptional regulatory element" /label="WPRE" polyA_signal 2935..2983 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 3185..3613 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3627..3956 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4023..4817 /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 4993..5114 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5163..5179) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5187..5203 /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(5211..5241) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5256..5277 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5565..6153) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6324..7184) /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(7185..7289) /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 taccgccacc atgggtgaaa ctctgggaga ttctcctgtt gacccagagc atggtgcctt 721 cgctgatgca ctgcctatga gcacttcaca agaaattacc atggttgaca cagagatgcc 781 attctggccc accaacttcg gaatcagctc tgtggacctc tccgtgatgg aagaccactc 841 gcattccttt gacatcaagc cctttaccac agttgatttc tccagcattt ctgctccaca 901 ctatgaagac attccattca caagagctga cccaatggtt gctgattaca aatatgacct 961 gaagctccaa gaataccaaa gtgcgatcaa agtagaacct gcatctccac cttattattc 1021 tgaaaagacc cagctctaca acaggcctca tgaagaacct tctaactccc tcatggccat 1081 tgagtgccga gtctgtgggg ataaagcatc aggcttccac tatggagttc atgcttgtga 1141 aggatgcaag ggttttttcc gaagaaccat ccgattgaag cttatttatg ataggtgtga 1201 tcttaactgc cggatccaca aaaaaagtag aaataaatgt cagtactgtc ggtttcagaa 1261 gtgccttgct gtggggatgt ctcacaatgc catcaggttt gggcggatgc cacaggccga 1321 gaaggagaag ctgttggcgg agatctccag tgatatcgac cagctgaacc cagagtctgc 1381 tgatctgcga gccctggcaa agcatttgta tgactcatac ataaagtcct tcccgctgac 1441 caaagccaag gcgagggcga tcttgacagg aaagacaacg gacaaatcac catttgtcat 1501 ctacgacatg aattccttaa tgatgggaga agataaaatc aagttcaaac atatcacccc 1561 cctgcaggag cagagcaaag aggtggccat ccgaattttt caagggtgcc agtttcgatc 1621 cgtagaagcc gtgcaagaga tcacagagta tgccaaaaat atccctggtt tcattaacct 1681 tgatttgaat gaccaagtga ctctgctcaa gtatggtgtc catgagatca tctacacgat 1741 gctggcctcc ctgatgaata aagatggagt cctcatctca gagggccaag gattcatgac 1801 cagggagttc ctcaaaagcc tgcggaagcc ctttggtgac tttatggagc ctaagtttga 1861 gtttgctgtg aagttcaatg cactggaatt agatgacagt gacttggcta tatttatagc 1921 tgtcattatt ctcagtggag accgcccagg cttgctgaac gtgaagccca tcgaggacat 1981 ccaagacaac ctgctgcagg ccctggaact gcagctcaag ctgaatcacc cagagtcctc 2041 tcagctgttc gccaaggtgc tccagaagat gacagacctc aggcagatcg tcacagagca 2101 cgtgcagcta ctgcatgtga tcaagaagac agagacagac atgagccttc accccctgct 2161 ccaggagatc tacaaggact tgtatctcga ggactacaaa gaccatgacg gtgattataa 2221 agatcatgac atcgactaca aggatgacga tgacaagtag tgagggcccg atatctcgac 2281 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct 2341 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt 2401 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg 2461 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact 2521 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct 2581 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg 2641 ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc atggctgctc 2701 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc 2761 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt 2821 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc tggaaacggg 2881 ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa 2941 taaaaagaca gaataaaacg cacgggtgtt gggtcgtttg ttcataaacg cggggttcgg 3001 tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc 3061 gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc 3121 aacgtcgggg cggcaggccc tgccatagca gatctgcgca gctggggctc tagggggtat 3181 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg 3241 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc 3301 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg gcatcccttt agggttccga 3361 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt 3421 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat 3481 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat 3541 ttataaggga ttttggggat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa 3601 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct 3661 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa 3721 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa 3781 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt 3841 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct 3901 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc 3961 tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc 4021 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat 4081 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt 4141 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac 4201 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg 4261 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc 4321 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa 4381 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc 4441 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg 4501 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg 4561 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa 4621 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg 4681 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct 4741 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc 4801 ttgacgagtt cttctgagcg ggactctggg gttcgcgaaa tgaccgacca agcgacgccc 4861 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga 4921 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc 4981 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc 5041 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc 5101 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca 5161 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga 5221 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt 5281 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga 5341 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc 5401 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg 5461 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc 5521 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc 5581 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga 5641 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc 5701 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa 5761 tgctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg 5821 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc 5881 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga 5941 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact 6001 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt 6061 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag 6121 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg 6181 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa 6241 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata 6301 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg 6361 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata 6421 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg 6481 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct 6541 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt 6601 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc 6661 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga 6721 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt 6781 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc 6841 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa 6901 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca 6961 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca 7021 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct 7081 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc 7141 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa 7201 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt 7261 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc 7321 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg 7381 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 7441 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 7501 ttagg //