LOCUS       sequence-115800-         969 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Cas9n (D10A nickase mutant) from S. pyogenes with 2A-Puro, and 
            cloning backbone for sgRNA (V2.0).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 969)
  AUTHORS   Ran FA, Hsu PD, Wright J, Agarwala V, Scott DA, Zhang F
  TITLE     Genome engineering using the CRISPR-Cas9 system.
  JOURNAL   Nat Protoc. 2013 Nov;8(11):2281-308. doi: 10.1038/nprot.2013.143. 
            Epub 2013 Oct 24.
  PUBMED    24157548
REFERENCE   2  (bases 1 to 969)
  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"; doi: 
            "10.1038/nprot.2013.143"; journalName: "Nat Protoc"; date: 
            "2013-11"; volume: "8"; issue: "11"; pages: "2281-308"
COMMENT     Sequence Label: pSpCas9n(BB)-2A-Puro (PX462) V2.0
FEATURES             Location/Qualifiers
     source          1..969
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      55..64
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             64..129
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             136..156
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 gtttaattac ctggagcacc tgcctgaaat cacttttttt caggttggac cggtgccacc
       61 atggactata aggaccacga cggagactac aaggatcatg atattgatta caaagacgat
      121 gacgataaga tggccccaaa gaagaagcgg aaggtcggta tccacggagt cccagcagcc
      181 gacaagaagt acagcatcgg cctggccatc ggcaccaact ctgtgggctg ggccgtgatc
      241 accgacgagt acaaggtgcc cagcaagaaa ttcaaggtgc tgggcaacac cgaccggcac
      301 agcatcaaga agaacctgat cggagccctg ctgttcgaca gcggcgaaac agccgaggcc
      361 acccggctga agagaaccgc cagaagaaga tacaccagac ggaagaaccg gatctgctat
      421 ctgcaagaga tcttcagcaa cgagatggcc aaggtggacg acagcttctt ccacagactg
      481 gaagagtcct tcctggtgga agaggataag aagcacgagc ggcaccccat cttcggcaac
      541 atcgtggacg aggtggccta ccacgagaag taccccacca tctaccacct gagaaagaaa
      601 ctggtggaca gcaccgacaa ggccgacctg cggctgatct atctggccct ggcccacatg
      661 atcaagttcc ggggccactt cctgatcgag ggcgacctga accccgacaa cagcgacgtg
      721 gacaagctgt tcatccagct ggtgcagacc tacaaccagc tgttcgagga aaaccccatc
      781 aacgccagcg gcgtggacgc caaggccatc ctgtctgcca gactgagcaa gagcagacgg
      841 ctggaaaatc tgatcgccca gctgcccggc gagaagaaga atggcctgtt cggcaacctg
      901 attgccctga gcctgggcct gacccccaac ttcaagagca acttcgacct ggccgaggat
      961 gccaaactg
//
