LOCUS       sequence-129998-         892 bp    DNA     linear   UNA 15-APR-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 892)
  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 892)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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..892
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    25..49
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      53..62
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             74..100
                     /codon_start=1
                     /product="nuclear localization signal (Makkerh et al., 
                     1996)"
                     /label=NLS
                     /translation="PAAKKKKLD"
     CDS             164..190
                     /codon_start=1
                     /product="nuclear localization signal (Makkerh et al., 
                     1996)"
                     /label=NLS
                     /translation="PAAKKKKLD"
ORIGIN
        1 ccatcgattc gaattcaagg atcaacaagt ttgtacaaaa aagcaggctg gcgccaccat
       61 gggtagagcc gcgccggcag ctaagaaaaa gaaactggat caattgggcg ctagcggtag
      121 tcccgggtct ggaggtagcg gctcaagtgg ccgtacggca gctcctgcgg caaaaaagaa
      181 aaagttggac tctgaattcg gaagcggaag cgacaagaag tactccattg ggctcgatat
      241 cggtaccaac agcgtcggct gggccgtcat tacggacgag tacaaggtgc cgagcaaaaa
      301 attcaaagtt ctgggcaata ccgatcgcca cagcataaag aagaacctca ttggagccct
      361 cctgttcgac tccggggaga cggccgaagc cacgcggctc aaaagaacag cacggcgcag
      421 atatacccgc agaaagaatc ggatctgcta cctgcaggag atctttagta atgagatggc
      481 taaggtggat gactctttct tccataggct ggaggagtcc tttttggtgg aggaggataa
      541 aaagcacgag cgccacccaa tctttggcaa tatcgtggac gaggtggcgt accatgaaaa
      601 gtacccaacc atatatcatc tgaggaagaa gctggtagac agtactgata aggctgactt
      661 gcggttgatc tatctcgcgc tggcgcacat gatcaaattt cggggacact tcctcatcga
      721 gggggacctg aacccagaca acagcgatgt cgacaaactc tttatccaac tggttcagac
      781 ttacaatcag cttttcgagg agaacccgat caacgcatcc ggcgttgacg ccaaagcaat
      841 cctgagcgct aggctgtcca aatcccggcg gctcgaaaac ctcatcgcac ag
//
