LOCUS Exported 4335 bp ds-DNA linear SYN 18-MAY-2021 DEFINITION encodes yeast-optimized dCas9_VP64 synthetic transcription factor. ACCESSION . VERSION . KEYWORDS pTPGI_dCas9_VP64 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4335) AUTHORS Farzadfard F, Perli SD, Lu TK TITLE Tunable and Multifunctional Eukaryotic Transcription Factors Based on CRISPR/Cas. JOURNAL ACS Synth Biol. 2013 Sep 11. PUBMED 23977949 REFERENCE 2 (bases 1 to 4335) AUTHORS . TITLE Direct Submission JOURNAL Exported May 18, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4335 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 13..33 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 43..4143 /codon_start=1 /product="catalytically dead mutant of the Cas9 endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system" /label=dCas9 /note="RNA-guided DNA-binding protein that lacks endonuclease activity due to the D10A mutation in the RuvC catalytic domain and the H840A mutation in the HNH catalytic domain" /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD ATLIHQSITGLYETRIDLSQLGGD" CDS 4180..4329 /codon_start=1 /product="tetrameric repeat of the minimal activation domain of herpes simplex virus VP16 (Beerli et al., 1998)" /label=VP64 /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF DLDML" ORIGIN 1 atgtctagag ccccaaagaa gaagagaaaa gttagacccg gggataagaa atactctatt 61 ggtttggcta tcggtacaaa ctctgttggt tgggctgtta ttactgatga atacaaggtt 121 ccatccaaga agttcaaggt tttgggtaac actgatagac actccatcaa aaagaacttg 181 attggtgcct tgttgttcga ttctggtgaa actgctgaag ctactagatt gaaaagaacc 241 gctagaagaa gatacaccag aagaaagaac agaatctgct acttgcaaga aatcttctcc 301 aacgaaatgg ccaaggttga tgattcattc ttccacagat tggaagaatc cttcttggtc 361 gaagaagata agaagcacga aagacatcca atcttcggta acatcgttga tgaagttgct 421 taccacgaaa agtacccaac tatctaccat ttgagaaaga agttggttga ctctaccgat 481 aaggctgatt tgagattgat ctatttggct ttggcccaca tgattaagtt cagaggtcat 541 ttcttgatcg aaggtgattt gaacccagat aactccgatg ttgataagtt gttcatccaa 601 ttagtccaaa cctacaatca attattcgaa gaaaacccaa tcaacgcctc tggtgttgat 661 gctaaagcta ttttgtctgc cagattgtcc aagtccagaa gattagaaaa tttgatcgcc 721 caattaccag gtgaaaagaa gaatggtttg ttcggtaatt tgattgcctt gtctttgggt 781 ttgactccaa acttcaagtc caatttcgat ttggctgaag atgccaagtt gcaattatct 841 aaggatacct acgatgacga tttggataac ttgttggctc aaatcggtga tcaatacgct 901 gatttgtttt tggctgctaa gaacttgtcc gatgccattt tgttgtccga tattttgaga 961 gtcaacaccg aaattactaa ggctccattg tctgcctcta tgatcaaaag atacgatgaa 1021 caccaccaag acttgacttt gttgaaggct ttggtcagac aacaattacc tgaaaagtac 1081 aaagaaattt tcttcgatca atccaagaac ggttacgccg gttatattga tggtggtgct 1141 tctcaagaag aattttacaa gttcatcaag ccaatcttgg aaaagatgga cggtactgaa 1201 gaattattgg tcaagttgaa cagagaagat ttgttgagaa agcaaagaac cttcgacaac 1261 ggttctattc cacatcaaat tcacttgggt gaattgcacg caattttgag aagacaagaa 1321 gatttttatc cattcttgaa ggacaacaga gaaaagatcg aaaagattct gaccttcaga 1381 atcccttact acgttggtcc attggctaga ggtaattcaa gatttgcctg gatgactaga 1441 aagtccgaag aaactattac tccttggaac ttcgaagaag ttgtagataa gggtgcttct 1501 gcccaatcct ttattgaaag aatgaccaac ttcgacaaga acttgccaaa cgaaaaggtt 1561 ttgccaaagc actctttgtt gtacgaatac ttcaccgtct acaacgaatt gactaaggtt 1621 aagtacgtca ccgaaggtat gagaaaacca gcttttttat ccggtgaaca aaagaaggct 1681 atcgtcgatt tgttgttcaa gaccaacaga aaggttactg tcaagcaatt aaaagaagat 1741 tacttcaaga aaatcgaatg cttcgactcc gttgaaattt ctggtgtcga agatagattc 1801 aatgcctctt taggtactta ccatgacttg ttgaaaatca tcaaggacaa ggatttcttg 1861 gacaacgaag aaaacgaaga tattttggaa gatattgtct tgacattgac cttgtttgaa 1921 gatagagaaa tgattgaaga aagattgaaa acctacgccc acttgttcga tgataaggtt 1981 atgaagcaat taaagagaag aagatacact ggttggggta gattgtccag aaaattgatt 2041 aacggtatca gagacaagca atccggtaag accattttgg actttttgaa gtctgatggt 2101 ttcgctaaca gaaacttcat gcaattaatc cacgacgatt ccttgacttt caaagaagat 2161 atacaaaagg cccaagtctc tggtcaaggt gattctttac atgaacatat cgctaacttg 2221 gctggttctc cagctattaa gaagggtatt ttacaaaccg ttaaggtcgt tgacgaattg 2281 gtcaaagtta tgggtagaca taagccagaa aacatcgtta tcgaaatggc tagagaaaat 2341 caaaccaccc aaaagggtca aaagaactcc agagaaagaa tgaagagaat cgaagaaggt 2401 atcaaagaat tgggttccca aattttgaaa gaacacccag ttgaaaacac ccaattacaa 2461 aacgaaaagt tgtacttgta ctacttgcaa aacggtagag atatgtacgt tgaccaagaa 2521 ttggacatca acagattgtc tgattacgat gttgacgcta tcgttccaca atcttttttg 2581 aaggatgact ccattgacaa caaggtcttg actagatccg ataagaatag aggtaagtcc 2641 gataacgttc catctgaaga agtcgttaag aaaatgaaga actattggag acaattattg 2701 aacgccaagt tgatcaccca aagaaagttt gacaatttga ccaaggctga aagaggtggt 2761 ttgtctgaat tggataaggc aggtttcatc aaaagacaat tagtagaaac cagacaaatc 2821 accaagcacg ttgctcaaat tttggatagt agaatgaaca ctaagtacga cgaaaacgac 2881 aaattgatca gagaagttaa ggtcattacc ttgaagtcca agttggtttc cgatttcaga 2941 aaggacttcc aattctacaa ggtcagagaa atcaacaact accatcatgc acatgatgct 3001 tacttgaatg ctgttgttgg tactgccttg attaagaagt atccaaagtt ggaatccgaa 3061 tttgtctacg gtgattacaa ggtttacgac gttagaaaga tgatcgccaa gtccgaacaa 3121 gaaattggta aagctactgc caaatacttc ttctactcca atattatgaa tttctttaag 3181 accgaaatca ctttggccaa cggtgaaatt agaaaaagac cattgattga aactaatggt 3241 gaaacaggtg aaatcgtttg ggataagggt agagattttg ccactgttag aaaggtattg 3301 tccatgccac aagtaaacat cgtcaaaaag accgaagttc aaactggtgg tttctccaaa 3361 gaatccattt tgcctaagag aaactccgat aagttgatcg ctagaaaaaa agactgggac 3421 ccaaaaaagt acggtggttt tgattctcca actgttgctt actctgtttt ggttgttgct 3481 aaggtcgaaa agggtaagag taagaagttg aagtccgtca aagaattatt aggtatcact 3541 atcatggaaa gatcctcatt cgaaaagaat cctatcgact ttttggaagc caagggttac 3601 aaagaagtca agaaggactt gatcattaag ttgccaaagt acagtttgtt cgaattggaa 3661 aatggtagaa agagaatgtt ggcttctgcc ggtgaattac aaaagggtaa tgaattggct 3721 ttgccatcca agtacgttaa tttcttatac ttggcctccc actacgaaaa attgaaaggt 3781 tctcctgaag ataacgaaca aaagcaatta tttgtcgaac aacacaagca ctacttggac 3841 gaaatcattg aacaaatttc cgaattttcc aaaagagtca ttttggctga cgccaatttg 3901 gacaaagttt tgtcagctta caacaagcac agagataagc caattagaga acaagctgaa 3961 aacatcattc acttgttcac tttgactaac ttgggtgctc cagctgcttt taagtatttc 4021 gataccacta tcgacagaaa gagatacacc tctaccaaag aagttttgga cgctactttg 4081 atccaccaat ctattactgg tttgtacgaa actagaatcg acttgtctca attaggtggt 4141 gatggttctg gtagatctgg agtcgacggt ggaggttctg acgctttgga cgacttcgac 4201 ttggatatgc tgggttctga tgcgctagat gactttgacc tcgacatgct tggaagtgac 4261 gccttagatg attttgacct ggatatgctt ggatcagacg ctctggacga tttcgactta 4321 gacatgcttt cctag //