LOCUS Exported 9189 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS YAP1 (S6A) - V5 in pLX304 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9189) AUTHORS Rosenbluh J, Nijhawan D, Cox AG, Li X, Neal JT, Schafer EJ, Zack TI, Wang X, Tsherniak A, Schinzel AC, Shao DD, Schumacher SE, Weir BA, Vazquez F, Cowley GS, Root DE, Mesirov JP, Beroukhim R, Kuo CJ, Goessling W, Hahn WC TITLE beta-Catenin-Driven Cancers Require a YAP1 Transcriptional Complex for Survival and Tumorigenesis. JOURNAL Cell. 2012 Dec 21;151(7):1457-73. doi: 10.1016/j.cell.2012.11.026. Epub 2012 Dec 13. PUBMED 23245941 REFERENCE 2 (bases 1 to 9189) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9189 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 1..304 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 305..508 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 459..479 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" protein_bind 531..555 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" protein_bind complement(2023..2047) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" CDS 2049..2090 /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" misc_feature 2132..2720 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(2185..2205) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(2603..2614) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 2792..3025 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 3103..3224 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3140..3159) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3194..3213 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin 3264..3399 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3326..3345 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind complement(3419..3438) /label=T7 /note="T7 promoter, forward primer" promoter complement(3420..3438) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(3448..3465) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(3448..3464) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3457..3479) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 3606..4061 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3693..3712) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3903..3924 /label=F1ori-F /note="F1 origin, forward primer" promoter 4087..4191 /gene="bla" /label=AmpR promoter CDS 4192..5052 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(4410..4429) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 5223..5811 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5712..5731 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 5965..5982 /label=L4440 /note="L4440 vector, forward primer" protein_bind 6099..6120 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 6135..6165 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6173..6189 /label=lac operator /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-beta-D-thiogalactopyranoside (IPTG)." primer_bind 6178..6200 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 6197..6213 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 6197..6213 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 6232..6252 /label=T3 /note="T3 promoter, forward primer" promoter 6234..6252 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" promoter 6280..6506 /label=RSV promoter /note="Rous sarcoma virus enhancer/promoter" LTR 6507..6687 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 6734..6859 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 7352..7585 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 7770..7814 /codon_start=1 /product="antigenic peptide corresponding to amino acids 655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik et al., 2013)" /label=gp41 peptide /note="recognized by the 2H10 single-chain llama nanobody" /translation="KNEQELLELDKWASL" promoter 7989..8489 /label=hPGK promoter /note="human phosphoglycerate kinase 1 promoter" primer_bind 8408..8426 /label=hPGK-F /note="Human PGK promoter, forward primer" CDS 8501..8896 /codon_start=1 /gene="Aspergillus terreus BSD" /product="blasticidin S deaminase" /label=BSD /note="confers resistance to blasticidin" /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI KAIVKDSDGQPTAVGIRELLPSGYVWEG" misc_feature 8956..9073 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" ORIGIN 1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt 61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca 121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc 181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta 241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac 301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg 361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg 421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt 481 acggtgggag gtctatataa gcagagctct ctggctaact gtcgggatca acaagtttgt 541 acaaaaaagt tggcatggat cccgggcagc agccgccgcc tcaaccggcc ccccagggcc 601 aagggcagcc gccttcgcag cccccgcagg ggcagggccc gccgtccgga cccgggcaac 661 cggcacccgc ggcgacccag gcggcgccgc aggcaccccc cgccgggcat cagatcgtgc 721 acgtccgcgg ggacgcggag accgacctgg aggcgctctt caacgccgtc atgaacccca 781 agacggccaa cgtgccccag accgtgccca tgaggctccg gaagctgccc gactccttct 841 tcaagccgcc ggagcccaaa tcccactccc gacaggccgc tactgatgca ggcactgcag 901 gagccctgac tccacagcat gttcgagctc atgccgctcc agctgctctg cagttgggag 961 ctgtttctcc tgggacactg acccccactg gagtagtctc tggcccagca gctacaccca 1021 cagctcagca tcttcgacag gctgcttttg agatacctga tgatgtacct ctgccagcag 1081 gttgggagat ggcaaagaca tcttctggtc agagatactt cttaaatcac atcgatcaga 1141 caacaacatg gcaggacccc aggaaggcca tgctgtccca gatgaacgtc acagccccca 1201 ccagtccacc agtgcagcag aatatgatga actcggcttc aggtcctctt cctgatggat 1261 gggaacaagc catgactcag gatggagaaa tttactatat aaaccataag aacaagacca 1321 cctcttggct agacccaagg cttgaccctc gttttgccat gaaccagaga atcagtcaga 1381 gtgctccagt gaaacagcca ccacccctgg ctccccagag cccacaggga ggcgtcatgg 1441 gtggcagcaa ctccaaccag cagcaacaga tgcgactgca gcaactgcag atggagaagg 1501 agaggctgcg gctgaaacag caagaactgc ttcggcagga gttagccctg cgtagccagt 1561 taccaacact ggagcaggat ggtgggactc aaaatccagt gtcttctccc gggatgtctc 1621 aggaattgag aacaatgacg accaatagct cagatccttt ccttaacagt ggcacctatc 1681 actctcgaga tgaggctaca gacagtggac taagcatgag cagctacagt gtccctcgaa 1741 ccccagatga cttcctgaac agtgtggatg agatggatac aggtgatact atcaaccaaa 1801 gcaccctgcc ctcacagcag aaccgtttcc cagactacct tgaagccatt cctgggacaa 1861 atgtggacct tggaacactg gaaggagatg gaatgaacat agaaggagag gagctgatgc 1921 caagtctgca ggaagctttg agttctgaca tccttaatga catggagtct gttttggctg 1981 ccaccaagct agataaagaa agctttctta catggttacc acccccaact ttcttgtaca 2041 aagtggttgg taagcctatc cctaaccctc tcctcggtct cgattctacg tagtaatgag 2101 ctagcgctaa ccggtggcgc gttaagtcga caatcaacct ctggattaca aaatttgtga 2161 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt 2221 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa 2281 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt 2341 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct 2401 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg 2461 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc 2521 ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg 2581 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct 2641 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc 2701 cctttgggcc gcctccccgc gtcgacttta agaccaatga cttacaaggc agctgtagat 2761 cttagccact ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaga 2821 caagatctgc tttttgcttg tactgggtct ctctggttag accagatctg agcctgggag 2881 ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt 2941 caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt 3001 tagtcagtgt ggaaaatctc tagcagtacg tatagtagtt catgtcatct tattattcag 3061 tatttataac ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt tattgcagct 3121 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca 3181 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggctctag 3241 ctatcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc 3301 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg 3361 agctattcca gaagtagtga ggaggctttt ttggaggcct agggacgtac ccaattcgcc 3421 ctatagtgag tcgtattacg cgcgctcact ggccgtcgtt ttacaacgtc gtgactggga 3481 aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg 3541 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga 3601 atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt 3661 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct 3721 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg 3781 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag 3841 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa 3901 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga 3961 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa 4021 atttaacgcg aattttaaca aaatattaac gcttacaatt taggtggcac ttttcgggga 4081 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc 4141 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt 4201 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct 4261 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt 4321 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt 4381 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac 4441 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac 4501 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct 4561 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg 4621 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg 4681 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca 4741 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa 4801 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt 4861 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc 4921 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg 4981 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt 5041 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt 5101 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc 5161 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct 5221 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta 5281 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc 5341 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac 5401 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct 5461 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat 5521 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg 5581 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa 5641 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg 5701 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga 5761 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc 5821 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct 5881 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct 5941 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca 6001 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg 6061 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat 6121 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc 6181 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagcg cgcaattaac 6241 cctcactaaa gggaacaaaa gctggagctg caagcttaat gtagtcttat gcaatactct 6301 tgtagtcttg caacatggta acgatgagtt agcaacatgc cttacaagga gagaaaaagc 6361 accgtgcatg ccgattggtg gaagtaaggt ggtacgatcg tgccttatta ggaaggcaac 6421 agacgggtct gacatggatt ggacgaacca ctgaattgcc gcattgcaga gatattgtat 6481 ttaagtgcct agctcgatac ataaacgggt ctctctggtt agaccagatc tgagcctggg 6541 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc 6601 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct 6661 tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac agggacttga aagcgaaagg 6721 gaaaccagag gagctctctc gacgcaggac tcggcttgct gaagcgcgca cggcaagagg 6781 cgaggggcgg cgactggtga gtacgccaaa aattttgact agcggaggct agaaggagag 6841 agatgggtgc gagagcgtca gtattaagcg ggggagaatt agatcgcgat gggaaaaaat 6901 tcggttaagg ccagggggaa agaaaaaata taaattaaaa catatagtat gggcaagcag 6961 ggagctagaa cgattcgcag ttaatcctgg cctgttagaa acatcagaag gctgtagaca 7021 aatactggga cagctacaac catcccttca gacaggatca gaagaactta gatcattata 7081 taatacagta gcaaccctct attgtgtgca tcaaaggata gagataaaag acaccaagga 7141 agctttagac aagatagagg aagagcaaaa caaaagtaag accaccgcac agcaagcggc 7201 cgctgatctt cagacctgga ggaggagata tgagggacaa ttggagaagt gaattatata 7261 aatataaagt agtaaaaatt gaaccattag gagtagcacc caccaaggca aagagaagag 7321 tggtgcagag agaaaaaaga gcagtgggaa taggagcttt gttccttggg ttcttgggag 7381 cagcaggaag cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc agacaattat 7441 tgtctggtat agtgcagcag cagaacaatt tgctgagggc tattgaggcg caacagcatc 7501 tgttgcaact cacagtctgg ggcatcaagc agctccaggc aagaatcctg gctgtggaaa 7561 gatacctaaa ggatcaacag ctcctgggga tttggggttg ctctggaaaa ctcatttgca 7621 ccactgctgt gccttggaat gctagttgga gtaataaatc tctggaacag atttggaatc 7681 acacgacctg gatggagtgg gacagagaaa ttaacaatta cacaagctta atacactcct 7741 taattgaaga atcgcaaaac cagcaagaaa agaatgaaca agaattattg gaattagata 7801 aatgggcaag tttgtggaat tggtttaaca taacaaattg gctgtggtat ataaaattat 7861 tcataatgat agtaggaggc ttggtaggtt taagaatagt ttttgctgta ctttctatag 7921 tgaatagagt taggcaggga tattcaccat tatcgtttca gacccacctc ccaaccccga 7981 ggggaccctt gcgccttttc caaggcagcc ctgggtttgc gcagggacgc ggctgctctg 8041 ggcgtggttc cgggaaacgc agcggcgccg accctgggtc tcgcacattc ttcacgtccg 8101 ttcgcagcgt cacccggatc ttcgccgcta cccttgtggg ccccccggcg acgcttcctg 8161 ctccgcccct aagtcgggaa ggttccttgc ggttcgcggc gtgccggacg tgacaaacgg 8221 aagccgcacg tctcactagt accctcgcag acggacagcg ccagggagca atggcagcgc 8281 gccgaccgcg atgggctgtg gccaatagcg gctgctcagc agggcgcgcc gagagcagcg 8341 gccgggaagg ggcggtgcgg gaggcggggt gtggggcggt agtgtgggcc ctgttcctgc 8401 ccgcgcggtg ttccgcattc tgcaagcctc cggagcgcac gtcggcagtc ggctccctcg 8461 ttgaccgaat caccgacctc tctccccagg gggtaccacc atggccaagc ctttgtctca 8521 agaagaatcc accctcattg aaagagcaac ggctacaatc aacagcatcc ccatctctga 8581 agactacagc gtcgccagcg cagctctctc tagcgacggc cgcatcttca ctggtgtcaa 8641 tgtatatcat tttactgggg gaccttgtgc agaactcgtg gtgctgggca ctgctgctgc 8701 tgcggcagct ggcaacctga cttgtatcgt cgcgatcgga aatgagaaca ggggcatctt 8761 gagcccctgc ggacggtgcc gacaggtgct tctcgatctg catcctggga tcaaagccat 8821 agtgaaggac agtgatggac agccgacggc agttgggatt cgtgaattgc tgccctctgg 8881 ttatgtgtgg gagggcctgc agctgcagta gtaagaattc tagatcttga gacaaatggc 8941 agtattcatc cacaatttta aaagaaaagg ggggattggg gggtacagtg caggggaaag 9001 aatagtagac ataatagcaa cagacataca aactaaagaa ttacaaaaac aaattacaaa 9061 aattcaaaat tttcgggttt attacaggga cagcagagat ccactttggc gccggctcga 9121 gggggcccgg ggttattaat agtaatcaat tacggggtca ttagttcata gcccatatat 9181 ggagttccg //