LOCUS       sequence-124565-         886 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses R1335K mutant SpCas9 fused to Zif268 in mammalian cell.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 886)
  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 886)
  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-Zif268
FEATURES             Location/Qualifiers
     source          1..886
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    39..63
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(365..391)
                     /codon_start=1
                     /product="nuclear localization signal (Makkerh et al., 
                     1996)"
                     /label=NLS
                     /translation="PAAKKKKLD"
     CDS             complement(404..493)
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     CDS             complement(494..514)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 atacgactca ctatagttct agaggctcga gaggatcaac cactttgtac aagaaagctg
       61 ggtctagata tctcgaggcg gccgcttagt ccttctgtct taaatggatt ttggtatgcc
      121 tcttgcgttc atcactcctg gcaaacttcc tcccacaaat gtcacaggca aaaggcttct
      181 cgcctgtgtg ggtgcggatg tgggtggtaa ggtggtcact acgactgaag ttacgcatgc
      241 agattcgaca ctggaagggc ttctgaccgg tatggatgcg gatatggcgg gtaagctcat
      301 ccgagcgaga aaagcggcga tcgcaggact cgacagggca agcatatggg cgtccggatt
      361 cgaaatccag tttctttttc ttagctgccg gagcggcgga tccagcgtaa tctggaacgt
      421 cataaggata cgatcctgca tagtccggga cgtcataggg atagcccgca tagtcaggaa
      481 catcgtatgg gtataccttc ctcttcttct tgggcccgcc ggtgccgtct ccaccgagct
      541 gagagaggtc gattcttgtt tcatagagcc ccgtaattga ctgatgaatc agtgtggcgt
      601 ccaggacctc ctttgtagag gtgtacttct ttctgtctat ggtggtgtcg aagtacttga
      661 aggctgcagg cgcgcccaag ttggtcagag taaacaagtg gataatgttt tctgcctgct
      721 ccctgatggg cttatccctg tgcttattgt aagcagaaag caccttatcg aggttagcgt
      781 cggcgaggat cactcttttg gagaactcgc ttatttgctc gatgatctca tcaaggtagt
      841 gtttgtgttg ttccacgaac agctgcttct gctcattatc ttcggg
//
