LOCUS       sequence-106485-        4992 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  ST1-dCas9 with VP64-p65-Rta (VPR) fused to it's C-terminus; 
            mammalian vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4992)
  AUTHORS   Chavez A, Scheiman J, Vora S, Pruitt BW, Tuttle M, P R Iyer E, Lin 
            S, Kiani S, Guzman CD, Wiegand DJ, Ter-Ovanesyan D, Braff JL, 
            Davidsohn N, Housden BE, Perrimon N, Weiss R, Aach J, Collins JJ, 
            Church GM
  TITLE     Highly efficient Cas9-mediated transcriptional programming.
  JOURNAL   Nat Methods. 2015 Mar 2. doi: 10.1038/nmeth.3312.
  PUBMED    25730490
REFERENCE   2  (bases 1 to 4992)
  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: "Nat 
            Methods. 2015 Mar 2. doi: 10.1038/nmeth.3312."
COMMENT     Sequence Label: M_ST1n_VPR
FEATURES             Location/Qualifiers
     source          1..4992
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             join(1..24,25..27,28..1791,1792..1797,1798..1863,1864..1866,
                     1867..3363)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus thermophilus Type II 
                     CRISPR/Cas system"
                     /label=St1dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the mutations D9A, D598A, 
                     H599A, and N622A"
                     /translation="MSDLVLGLAIGIGSVGVGILNKVTGEIIHKNSRIFPAAQAENNLV
                     RRTNRQGRRLARRKKHRRVRLNRLFEESGLITDFTKISINLNPYQLRVKGLTDELSNEE
                     LFIALKNMVKHRGISYLDDASDDGNSSVGDYAQIVKENSKQLETKTPGQIQLERYQTYG
                     QLRGDFTVEKDGKKHRLINVFPTSAYRSEALRILQTQQEFNPQITDEFINRYLEILTGK
                     RKYYHGPGNEKSRTDYGRYRTSGETLDNIFGILIGKCTFYPDEFRAAKASYTAQEFNLL
                     NDLNNLTVPTETKKLSKEQKNQIINYVKNEKAMGPAKLFKYIAKLLSCDVADIKGYRID
                     KSGKAEIHTFEAYRKMKTLETLDIEQMDRETLDKLAYVLTLNTEREGIQEALEHEFADG
                     SFSQKQVDELVQFRKANSSIFGKGWHNFSVKLMMELIPELYETSEEQMTILTRLGKQKT
                     TSSSNKTKYIDEKLLTEEIYNPVVAKSVRQAIKIVNAAIKEYGDFDNIVIEMARETNED
                     DEKKAIQKIQKANKDEKDAAMLKAANQYNGKAELPHSVFHGHKQLATKIRLWHQQGERC
                     LYTGKTISIHDLINNSNQFEVAAILPLSITFDDSLANKVLVYATAAQEKGQRTPYQALD
                     SMDDAWSFRELKAFVRESKTLSNKKKEYLLTEEDISKFDVRKKFIERNLVDTRYASRVV
                     LNALQEHFRAHKIDTKVSVVRGQFTSQLRRHWGIEKTRDTYHHHAVDALIIAASSQLNL
                     WKKQKNTLVSYSEDQLLDIETGELISDDEYKESVFKAPYQHFVDTLKSKEFEDSILFSY
                     QVDSKFNRKISDATIYATRQAKVGKDKADETYVLGKIKDIYTQDGYDAFMKIYKKDKSK
                     FLMYRHDPQTFEKVIEPILENYPNKQINDKGKEVPCNPFLKYKEEHGYIRKYSKKGNGP
                     EIKSLKYYDSKLGNHIDITPKDSNNKVVLQSVSPWRADVYFNKTTGKYEILGLKYADLQ
                     FDKGTGTYKISQEKYNDIKKKEGVDSDSEFKFTLYKNDLLLVKDTETKEQQLFRFLSRT
                     MPKQKHYVELKPYDKQKFEGGEALIKVLGNVANSGQCKKGLGKSNISIYKVRTDVLGNQ
                     HIIKNEGDKPKLDF"
     CDS             3376..3396
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3421..3570
                     /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"
     CDS             3595..3615
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4045..4401
                     /codon_start=1
                     /product="transcriptional activation domain of human RelA, 
                     also known as p65 (O'Shea and Perkins, 2008)"
                     /label=RelA (p65) AD
                     /translation="PTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNS
                     EFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGD
                     EDFSSIADMDFSALL"
     CDS             4420..4989
                     /codon_start=1
                     /product="transcriptional activation domain from the human 
                     herpesvirus 4 (Epstein-Barr virus) replication and 
                     transcription activator Rta/BRLF1 (Hardwick et al., 1992; 
                     Chavez et al., 2015)"
                     /label=Rta AD
                     /translation="RDSREGMFLPKPEAGSAISDVFEGREVCQPKRIRPFHPPGSPWAN
                     RPLPASLAPTPTGPVHEPVGSLTPAPVPQPLDPAPAVTPEASHLLEDPDEETSQAVKAL
                     REMADTVIPQKEEAAICGQMDLSHPPPRGHLDELTTTLESMTEDLNLDSPLTPELNEIL
                     DTFLNDECLLHAMHISTGLSIFDTSLF"
ORIGIN
        1 atgagcgacc tggtgctggg cctggccatc ggcatcggca gcgtgggcgt gggcatcctg
       61 aacaaggtga ccggcgagat catccacaag aacagtcgca tcttccctgc tgctcaggct
      121 gagaacaacc tggtgcgccg caccaaccgc cagggtcgcc ggcttgctcg ccgcaagaag
      181 caccggcgcg tgcgcctgaa ccgcctgttc gaggagagcg gcctgatcac cgacttcacc
      241 aagatcagca tcaacctgaa cccctaccag ctgcgcgtga agggcctgac cgacgagctg
      301 agcaacgagg agctgttcat cgccctgaag aacatggtga agcaccgcgg catcagctac
      361 ctggacgacg ccagcgacga cggcaacagc agcgtgggcg actacgccca gatcgtgaag
      421 gagaacagca agcagctgga gaccaagacc cccggccaga tccagctgga gcgctaccag
      481 acctacggcc agctgcgcgg cgacttcacc gtggagaagg acggcaagaa gcaccgcctg
      541 atcaacgtgt tccccaccag cgcctaccgc agcgaggccc tgcgcatcct gcagacccag
      601 caggagttca acccccagat caccgacgag ttcatcaacc gctacctgga gatcctgacc
      661 ggcaagcgca agtactacca cggccccggc aacgagaaga gccgcaccga ctacggccgc
      721 taccgcacca gcggcgagac cctggacaac atcttcggca tcctgatcgg caagtgcacc
      781 ttctaccccg acgagttccg cgccgccaag gccagctaca ccgcccagga gttcaacctg
      841 ctgaacgacc tgaacaacct gaccgtgccc accgagacca agaagctgag caaggagcag
      901 aagaaccaga tcatcaacta cgtgaagaac gagaaggcca tgggccccgc caagctgttc
      961 aagtacatcg ccaagctgct gagctgcgac gtggccgaca tcaagggcta ccgcatcgac
     1021 aagagcggca aggccgagat ccacaccttc gaggcctacc gcaagatgaa gaccctggag
     1081 accctggaca tcgagcagat ggaccgcgag accctggaca agctggccta cgtgctgacc
     1141 ctgaacaccg agcgcgaggg catccaggag gccctggagc acgagttcgc cgacggcagc
     1201 ttcagccaga agcaggtgga cgagctggtg cagttccgca aggccaacag cagcatcttc
     1261 ggcaagggct ggcacaactt cagcgtgaag ctgatgatgg agctgatccc cgagctgtac
     1321 gagaccagcg aggagcagat gaccatcctg acccgcctgg gcaagcagaa gaccaccagc
     1381 agcagcaaca agaccaagta catcgacgag aagctgctga ccgaggagat ctacaacccc
     1441 gtggtggcca agagcgtgcg ccaggccatc aagatcgtga acgccgccat caaggagtac
     1501 ggcgacttcg acaacatcgt gatcgagatg gcccgcgaga ccaacgagga cgacgagaag
     1561 aaggccatcc agaagatcca gaaggccaac aaggacgaga aggacgccgc catgctgaag
     1621 gccgccaacc agtacaacgg caaggccgag ctgccccaca gcgtgttcca cggccacaag
     1681 cagctggcca ccaagatccg cctgtggcac cagcagggcg agcgctgcct gtacaccggc
     1741 aagaccatca gcatccacga cctgatcaac aacagcaacc agttcgaggt ggctgccatc
     1801 ctgcccctga gcatcacctt cgacgacagc ctggccaaca aggtgctggt gtacgccacc
     1861 gccgctcagg agaagggcca gcgcaccccc taccaggccc tggacagcat ggacgacgcc
     1921 tggagcttcc gcgagctgaa ggccttcgtg cgcgagagca agaccctgag caacaagaag
     1981 aaggagtacc tgctgaccga ggaggacatc agcaagttcg acgtgcgcaa gaagttcatc
     2041 gagcgcaacc tggtggacac ccgctacgcc agccgcgtgg tgctgaacgc cctgcaggag
     2101 cacttccgcg cccacaagat cgacaccaag gtgagcgtgg tgcgcggcca gttcaccagc
     2161 cagctgcgcc gccactgggg catcgagaag acccgcgaca cctaccacca ccacgccgtg
     2221 gacgccctga tcattgcggc ttctagccag ctgaacctgt ggaagaagca gaagaacacc
     2281 ctggtgagct acagcgagga ccagctgctg gacatcgaga ccggcgagct gatcagcgac
     2341 gacgagtaca aggagagcgt gttcaaggcc ccctaccagc acttcgtgga caccctgaag
     2401 agcaaggagt tcgaggacag catcctgttc agctaccagg tggacagcaa gttcaaccgc
     2461 aagatcagcg acgccaccat ctacgccacc cgccaggcca aggtgggcaa ggacaaggcc
     2521 gacgagacct acgtgctggg caagatcaag gacatctaca cccaggacgg ctacgacgcc
     2581 ttcatgaaga tctacaagaa ggacaagagc aagttcctga tgtaccgcca cgacccccag
     2641 accttcgaga aggtgatcga gcccatcctg gagaactacc ccaacaagca gatcaacgat
     2701 aaaggcaagg aggtgccctg caaccccttc ctgaagtaca aggaggagca cggctacatc
     2761 cgcaagtaca gcaagaaggg caacggcccc gagatcaaga gcctgaagta ctacgacagc
     2821 aagctgggca accacatcga catcaccccc aaggacagca acaacaaggt ggtgctgcag
     2881 agcgtgagcc cctggcgcgc cgacgtgtac ttcaacaaga ccaccggcaa gtacgagatc
     2941 ctgggcctga agtacgccga cctgcagttt gataagggca ccggcaccta caagatcagc
     3001 caggagaagt acaacgacat caagaagaag gagggcgtgg acagcgacag cgagttcaag
     3061 ttcaccctgt acaagaacga ccttctgctg gtgaaggaca ccgagaccaa ggagcaacag
     3121 ctgttccgct tcctgagccg caccatgccc aagcagaagc actacgtgga gctgaagccc
     3181 tacgacaagc agaagttcga gggcggcgag gccctgatca aggtgctggg caacgtggcc
     3241 aacagcggcc agtgcaagaa gggcctgggc aagagcaaca tcagcatcta caaggtgcgc
     3301 accgacgtgc tgggcaacca gcacatcatc aagaacgagg gcgacaagcc caagctggac
     3361 ttcagcaggg ctgaccccaa gaagaagagg aaggtggagg ccagcggttc cggacgggct
     3421 gacgcattgg acgattttga tctggatatg ctgggaagtg acgccctcga tgattttgac
     3481 cttgacatgc ttggttcgga tgcccttgat gactttgacc tcgacatgct cggcagtgac
     3541 gcccttgatg atttcgacct ggacatgctg attaactcta gaagttccgg atctccgaaa
     3601 aagaaacgca aagttggtag ccagtacctg cccgacaccg acgaccggca ccggatcgag
     3661 gaaaagcgga agcggaccta cgagacattc aagagcatca tgaagaagtc ccccttcagc
     3721 ggccccaccg accctagacc tccacctaga agaatcgccg tgcccagcag atccagcgcc
     3781 agcgtgccaa aacctgcccc ccagccttac cccttcacca gcagcctgag caccatcaac
     3841 tacgacgagt tccctaccat ggtgttcccc agcggccaga tctctcaggc ctctgctctg
     3901 gctccagccc ctcctcaggt gctgcctcag gctcctgctc ctgcaccagc tccagccatg
     3961 gtgtctgcac tggctcaggc accagcaccc gtgcctgtgc tggctcctgg acctccacag
     4021 gctgtggctc caccagcccc taaacctaca caggccggcg agggcacact gtctgaagct
     4081 ctgctgcagc tgcagttcga cgacgaggat ctgggagccc tgctgggaaa cagcaccgat
     4141 cctgccgtgt tcaccgacct ggccagcgtg gacaacagcg agttccagca gctgctgaac
     4201 cagggcatcc ctgtggcccc tcacaccacc gagcccatgc tgatggaata ccccgaggcc
     4261 atcacccggc tcgtgacagg cgctcagagg cctcctgatc cagctcctgc ccctctggga
     4321 gcaccaggcc tgcctaatgg actgctgtct ggcgacgagg acttcagctc tatcgccgat
     4381 atggatttct cagccttgct gggtagcgga tcgggatctc gggattccag ggaagggatg
     4441 tttttgccga agcctgaggc cggctccgct attagtgacg tgtttgaggg ccgcgaggtg
     4501 tgccagccaa aacgaatccg gccatttcat cctccaggaa gtccatgggc caaccgccca
     4561 ctccccgcca gcctcgcacc aacaccaacc ggtccagtac atgagccagt cgggtcactg
     4621 accccggcac cagtccctca gccactggat ccagcgcccg cagtgactcc cgaggccagt
     4681 cacctgttgg aggatcccga tgaagagacg agccaggctg tcaaagccct tcgggagatg
     4741 gccgatactg tgattcccca gaaggaagag gctgcaatct gtggccaaat ggacctttcc
     4801 catccgcccc caaggggcca tctggatgag ctgacaacca cacttgagtc catgaccgag
     4861 gatctgaacc tggactcacc cctgaccccg gaattgaacg agattctgga taccttcctg
     4921 aacgacgagt gcctcttgca tgccatgcat atcagcacag gactgtccat cttcgacaca
     4981 tctctgtttt ga
//
