LOCUS       sequence-124481-         884 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses R1333K mutant SpCas9 in mammalian cell.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 884)
  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 884)
  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-MT1-NLS-3XHA-NLS
FEATURES             Location/Qualifiers
     source          1..884
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    36..60
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(98..124)
                     /codon_start=1
                     /product="nuclear localization signal (Makkerh et al., 
                     1996)"
                     /label=NLS
                     /translation="PAAKKKKLD"
     CDS             complement(137..226)
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     CDS             complement(227..247)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 cgactcacta tagttctaga ggctcgagag gatcaaccac tttgtacaag aaagctgggt
       61 ctagatatct cgaggcggcc gcttatccgg attcgaaatc cagtttcttt ttcttagctg
      121 ccggagcggc ggatccagcg taatctggaa cgtcataagg atacgatcct gcatagtccg
      181 ggacgtcata gggatagccc gcatagtcag gaacatcgta tgggtatacc ttcctcttct
      241 tcttgggccc gccggtgccg tctccaccga gctgagagag gtcgattctt gtttcataga
      301 gccccgtaat tgactgatga atcagtgtgg cgtccaggac ctcctttgta gaggtgtacc
      361 gctttttgtc tatggtggtg tcgaagtact tgaaggctgc aggcgcgccc aagttggtca
      421 gagtaaacaa gtggataatg ttttctgcct gctccctgat gggcttatcc ctgtgcttat
      481 tgtaagcaga aagcacctta tcgaggttag cgtcggcgag gatcactctt ttggagaact
      541 cgcttatttg ctcgatgatc tcatcaaggt agtgtttgtg ttgttccacg aacagctgct
      601 tctgctcatt atcttcggga gaccctttga gcttttcata gtggctggcc agatacaaga
      661 aattaacgta tttagagggc agtgccagct cgttaccttt ctgcagctcg cccgcactag
      721 cgagcattcg tttccggccg ttttcaagct caaagagaga gtacttgggc agcttaatga
      781 tgaggtcttt tttgacctct ttatatcctt tcgcttcgag aaagtcgatg gggtttttct
      841 cgaagctgga tcgctccatg attgtgatgc ccagcagttc cttg
//
