LOCUS       sequence-78157-e        1101 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Cas9n (D10A nickase mutant) from S. pyogenes with 2A-EGFP, and 
            cloning backbone for sgRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1101)
  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 1101)
  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-GFP (PX461)
FEATURES             Location/Qualifiers
     source          1..1101
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(288..341)
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             complement(357..404)
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
ORIGIN
        1 cgctcctgga cgtagccttc gggcatggcg gacttgaaga agtcgtgctg cttcatgtgg
       61 tcggggtagc ggctgaagca ctgcacgccg taggtcaggg tggtcacgag ggtgggccag
      121 ggcacgggca gcttgccggt ggtgcagatg aacttcaggg tcagcttgcc gtaggtggca
      181 tcgccctcgc cctcgccgga cacgctgaac ttgtggccgt ttacgtcgcc gtccagctcg
      241 accaggatgg gcaccacccc ggtgaacagc tcctcgccct tgctcactgg gccaggattc
      301 tcctcgacgt caccgcatgt tagcagactt cctctgccct ctccactgcc gaattccttt
      361 ttcttttttg cctggccggc ctttttcgtg gccgccggcc ttttgtcgcc tcccagctga
      421 gacaggtcga tccgtgtctc gtacaggccg gtgatgctct ggtggatcag ggtggcgtcc
      481 agcacctctt tggtgctggt gtacctcttc cggtcgatgg tggtgtcaaa gtacttgaag
      541 gcggcagggg ctcccagatt ggtcagggta aacaggtgga tgatattctc ggcctgctct
      601 ctgatgggct tatcccggtg cttgttgtag gcggacagca ctttgtccag attagcgtcg
      661 gccaggatca ctctcttgga gaactcgctg atctgctcga tgatctcgtc caggtagtgc
      721 ttgtgctgtt ccacaaacag ctgtttctgc tcattatcct cgggggagcc cttcagcttc
      781 tcatagtggc tggccaggta caggaagttc acatatttgg agggcagggc cagttcgttt
      841 cccttctgca gttcgccggc agaggccagc attctcttcc ggccgttttc cagctcgaac
      901 agggagtact taggcagctt gatgatcagg tcctttttca cttctttgta gcccttggct
      961 tccagaaagt cgatgggatt cttctcgaag ctgcttcttt ccatgatggt gatccccagc
     1021 agctctttca cactcttcag tttcttggac ttgccctttt ccactttggc caccaccagc
     1081 acagaatagg ccacggtggg g
//
