LOCUS       sequence-106842-         617 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 617)
  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 617)
  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..617
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(322..342)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(349..414)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     regulatory      414..423
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 tctgtggaag aagctgtcgt ccaccttggc catctcgttg ctgaagatct cttgcagata
       61 gcagatccgg ttcttccgtc tggtgtatct tcttctggcg gttctcttca gccgggtggc
      121 ctcggctgtt tcgccgctgt cgaacagcag ggctccgatc aggttcttct tgatgctgtg
      181 ccggtcggtg ttgcccagca ccttgaattt cttgctgggc accttgtact cgtcggtgat
      241 cacggcccag cccacagagt tggtgccgat ggccaggccg atgctgtact tcttgtcggc
      301 tgctgggact ccgtggatac cgaccttccg cttcttcttt ggggccatct tatcgtcatc
      361 gtctttgtaa tcaatatcat gatccttgta gtctccgtcg tggtccttat agtccatggt
      421 ggcaccggtc caacctgaaa aaaagtgatt tcaggcaggt gctccaggta attaaacatt
      481 aataccccac caaccaacca tcccttaaac ccttacctct tgctcagcta attacagccc
      541 ggaggagaag ggccgtcccg cccgctcacc tgtgggagta acgcggtcag tcagagccgg
      601 ggcgggcggc gcgaggc
//
