LOCUS       sequence-130001-         884 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  R1335K mutant SpCas9 expression plasmid to clone TALEs through BbsI 
            site (generates compatible overhangs with Acc65I and BamHI sites).
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: pCSDest2-2XNLS-SpCas9-MT3-NLS-3XHA-NLS-TALentry
FEATURES             Location/Qualifiers
     source          1..884
                     /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(149..175)
                     /codon_start=1
                     /product="nuclear localization signal (Makkerh et al., 
                     1996)"
                     /label=NLS
                     /translation="PAAKKKKLD"
     CDS             complement(188..277)
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     CDS             complement(278..298)
                     /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 gccgcttatc cggattaaga tcttgtcttc agctatacta
      121 gcatgaagac ttgtactccg gattcgaaat ccagtttctt tttcttagct gccggagcgg
      181 cggatccagc gtaatctgga acgtcataag gatacgatcc tgcatagtcc gggacgtcat
      241 agggatagcc cgcatagtca ggaacatcgt atgggtatac cttcctcttc ttcttgggcc
      301 cgccggtgcc gtctccaccg agctgagaga ggtcgattct tgtttcatag agccccgtaa
      361 ttgactgatg aatcagtgtg gcgtccagga cctcctttgt agaggtgtac ttctttctgt
      421 ctatggtggt gtcgaagtac ttgaaggctg caggcgcgcc caagttggtc agagtaaaca
      481 agtggataat gttttctgcc tgctccctga tgggcttatc cctgtgctta ttgtaagcag
      541 aaagcacctt atcgaggtta gcgtcggcga ggatcactct tttggagaac tcgcttattt
      601 gctcgatgat ctcatcaagg tagtgtttgt gttgttccac gaacagctgc ttctgctcat
      661 tatcttcggg agaccctttg agcttttcat agtggctggc cagatacaag aaattaacgt
      721 atttagaggg cagtgccagc tcgttacctt tctgcagctc gcccgcacta gcgagcattc
      781 gtttccggcc gttttcaagc tcaaagagag agtacttggg cagcttaatg atgaggtctt
      841 ttttgacctc tttatatcct ttcgcttcga gaaagtcgat gggg
//
