LOCUS V031406 6976 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V031406 VERSION V031406 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers enhancer 235..614 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 615..818 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" promoter 863..881 /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" CDS 932..2455 /label="YAP1(NM_001282101)" /note="YAP1(NM_001282101)" /gene="YAP1" CDS 2477..2542 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 2571..2795 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2841..3269 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3283..3612 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3679..4473 /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 4649..4770 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4819..4835) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4843..4859 /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(4867..4897) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4912..4933 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5221..5809) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5980..6840) /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(6841..6945) /gene="bla" /label="AmpR promoter" ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg 61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt 481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca 841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagt 901 taagcttggt accgagctcg gatccgccac catggatccc gggcagcagc cgccgcctca 961 accggccccc cagggccaag ggcagccgcc ttcgcagccc ccgcaggggc agggcccgcc 1021 gtccggaccc gggcaaccgg cacccgcggc gacccaggcg gcgccgcagg caccccccgc 1081 cgggcatcag atcgtgcacg tccgcgggga ctcggagacc gacctggagg cgctcttcaa 1141 cgccgtcatg aaccccaaga cggccaacgt gccccagacc gtgcccatga ggctccggaa 1201 gctgcccgac tccttcttca agccgccgga gcccaaatcc cactcccgac aggccagtac 1261 tgatgcaggc actgcaggag ccctgactcc acagcatgtt cgagctcatt cctctccagc 1321 ttctctgcag ttgggagctg tttctcctgg gacactgacc cccactggag tagtctctgg 1381 cccagcagct acacccacag ctcagcatct tcgacagtct tcttttgaga tacctgatga 1441 tgtacctctg ccagcaggtt gggagatggc aaagacatct tctggtcaga gatacttctt 1501 aaatcacatc gatcagacaa caacatggca ggaccccagg aaggccatgc tgtcccagat 1561 gaacgtcaca gcccccacca gtccaccagt gcagcagaat atgatgaact cggcttcagg 1621 tcctcttcct gatggatggg aacaagccat gactcaggat ggagaaattt actatataaa 1681 ccataagaac aagaccacct cttggctaga cccaaggctt gaccctcgtt ttgccatgaa 1741 ccagagaatc agtcagagtg ctccagtgaa acagccacca cccctggctc cccagagccc 1801 acagggaggc gtcatgggtg gcagcaactc caaccagcag caacagatgc gactgcagca 1861 actgcagatg gagaaggaga ggctgcggct gaaacagcaa gaactgcttc ggcaggtgag 1921 gccacaggca atgcggaata tcaatcccag cacagcaaat tctccaaaat gtcaggagtt 1981 agccctgcgt agccagttac caacactgga gcaggatggt gggactcaaa atccagtgtc 2041 ttctcccggg atgtctcagg aattgagaac aatgacgacc aatagctcag atcctttcct 2101 taacagtggc acctatcact ctcgagatga gagtacagac agtggactaa gcatgagcag 2161 ctacagtgtc cctcgaaccc cagatgactt cctgaacagt gtggatgaga tggatacagg 2221 tgatactatc aaccaaagca ccctgccctc acagcagaac cgtttcccag actaccttga 2281 agccattcct gggacaaatg tggaccttgg aacactggaa ggagatggaa tgaacataga 2341 aggagaggag ctgatgccaa gtctgcagga agctttgagt tctgacatcc ttaatgacat 2401 ggagtctgtt ttggctgcca ccaagctaga taaagaaagc tttcttacat ggttactcga 2461 gtctagaggg cccttcgact acaaagacca tgacggtgat tataaagatc atgacatcga 2521 ctacaaggat gacgatgaca agtgagttta aacccgctga tcagcctcga ctgtgccttc 2581 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc 2641 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg 2701 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa 2761 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg 2821 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt 2881 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt 2941 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct 3001 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg 3061 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga 3121 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc 3181 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga 3241 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt 3301 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca 3361 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat 3421 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg 3481 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc 3541 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta 3601 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca 3661 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct 3721 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc 3781 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc 3841 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc 3901 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg 3961 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc 4021 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac 4081 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat 4141 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc 4201 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg 4261 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg 4321 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc 4381 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc 4441 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gcgaaatgac 4501 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga 4561 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga 4621 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa 4681 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg 4741 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta 4801 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat 4861 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag 4921 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg 4981 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc 5041 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc 5101 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa 5161 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt 5221 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg 5281 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg 5341 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag 5401 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc 5461 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa 5521 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg 5581 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc 5641 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac 5701 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg 5761 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt 5821 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt 5881 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa 5941 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga 6001 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt 6061 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg 6121 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga 6181 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga 6241 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg 6301 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc 6361 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc 6421 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca 6481 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac 6541 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg 6601 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc 6661 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg 6721 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac 6781 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat 6841 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata 6901 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa 6961 agtgccacct gacgtc //