LOCUS       sequence-73283-e        1053 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 1053)
  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 1053)
  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..1053
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(36..89)
                     /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(105..152)
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
ORIGIN
        1 accaccccgg tgaacagctc ctcgcccttg ctcactgggc caggattctc ctcgacgtca
       61 ccgcatgtta gcagacttcc tctgccctct ccactgccga attccttttt cttttttgcc
      121 tggccggcct ttttcgtggc cgccggcctt ttgtcgcctc ccagctgaga caggtcgatc
      181 cgtgtctcgt acaggccggt gatgctctgg tggatcaggg tggcgtccag cacctctttg
      241 gtgctggtgt acctcttccg gtcgatggtg gtgtcaaagt acttgaaggc ggcaggggct
      301 cccagattgg tcagggtaaa caggtggatg atattctcgg cctgctctct gatgggctta
      361 tcccggtgct tgttgtaggc ggacagcact ttgtccagat tagcgtcggc caggatcact
      421 ctcttggaga actcgctgat ctgctcgatg atctcgtcca ggtagtgctt gtgctgttcc
      481 acaaacagct gtttctgctc attatcctcg ggggagccct tcagcttctc atagtggctg
      541 gccaggtaca ggaagttcac atatttggag ggcagggcca gttcgtttcc cttctgcagt
      601 tcgccggcag aggccagcat tctcttccgg ccgttttcca gctcgaacag ggagtactta
      661 ggcagcttga tgatcaggtc ctttttcact tctttgtagc ccttggcttc cagaaagtcg
      721 atgggattct tctcgaagct gcttctttcc atgatggtga tccccagcag ctctttcaca
      781 ctcttcagtt tcttggactt gcccttttcc actttggcca ccaccagcac agaataggcc
      841 acggtggggc tgtcgaagcc gccgtacttc ttagggtccc agtccttctt tctggcgatc
      901 agcttatcgc tgttcctctt gggcaggata gactctttgc tgaagccgcc tgtctgcacc
      961 tcggtctttt tcacgatatt cacttggggc atgctcagca ctttccgcac ggtggcaaaa
     1021 tcccggccct tatcccacac gatctccccg gtt
//
