LOCUS       sequence-125725-         920 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses R1335K mutant SpCas9 fused to ZFP-TS2 in mammalian cell.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 920)
  AUTHORS   Bolukbasi MF, Gupta A, Oikemus S, Derr AG, Garber M, Brodsky MH, Zhu
            LJ, Wolfe SA
  TITLE     DNA-binding-domain fusions enhance the targeting range and precision
            of Cas9.
  JOURNAL   Nat Methods. 2015 Oct 19. doi: 10.1038/nmeth.3624.
  PUBMED    26480473
REFERENCE   2  (bases 1 to 920)
  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 Oct 19. doi: 10.1038/nmeth.3624."
COMMENT     Sequence Label: pCSDest2-SpCas9-MT3-NLS-3XHA-NLS-ZFP_TS2
FEATURES             Location/Qualifiers
     source          1..920
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     polyA_signal    88..222
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(125..144)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     179..198
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(632..658)
                     /codon_start=1
                     /product="nuclear localization signal (Makkerh et al., 
                     1996)"
                     /label=NLS
                     /translation="PAAKKKKLD"
     CDS             complement(671..760)
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     CDS             complement(761..781)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 atgcattggc gccgcggccg cgaattaaaa aacctcccac acctccccct gaacctgaaa
       61 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa
      121 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
      181 ggtttgtcca aactcatcaa tgtatcttat catgtctgga tctacgtaat acgactcact
      241 atagttctag aggctcgagt cattacccgg ggtctttctg acggaggtgg atcttggtgt
      301 gccgtgtcag gtcgtcgctc cgcgcgaact tcctgccgca gatgtcacag gcgaagggct
      361 tttctcctgt gtgggtgcgg atgtgccggg tcaggtggcc cttgtcgctg aagttcctca
      421 tgcagatgcg gcattgaaac ggtttttgcc ctgtgtgaat gcggatgtgc tgtgtcaggt
      481 ggtcgctccg actgaagcgg cggtcacagg actcgacagg acacgcgtat ggcttctcac
      541 cagtgtgcgt gcgttggtgc cgtgtcaggt ggctcttctc cgagaaggac ttgccacatt
      601 caggacattt atagggcttt ccggattcga aatccagttt ctttttctta gctgccggag
      661 cggcggatcc agcgtaatct ggaacgtcat aaggatacga tcctgcatag tccgggacgt
      721 catagggata gcccgcatag tcaggaacat cgtatgggta taccttcctc ttcttcttgg
      781 gcccgccggt gccgtctcca ccgagctgag agaggtcgat tcttgtttca tagagccccg
      841 taattgactg atgaatcagt gtggcgtcca ggacctcctt tgtagaggtg tacttctttc
      901 tgtctatggt ggtgtcgaag
//
