LOCUS       sequence-71930-e        4335 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  encodes yeast-optimized dCas9_VP64 synthetic transcription factor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
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 Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "ACS Synth 
            Biol. 2013 Sep 11."
COMMENT     Sequence Label: pTPGI_dCas9_VP64
FEATURES             Location/Qualifiers
     source          1..4335
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             13..33
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(43..66,67..69,70..2556,2557..2559,2560..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
//
