LOCUS       sequence-99234-e        1151 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 1151)
  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 1151)
  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..1151
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(289..342)
                     /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(358..405)
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
ORIGIN
        1 gcgctcctgg acgtagcctt cgggcatggc ggacttgaag aagtcgtgct gcttcatgtg
       61 gtcggggtag cggctgaagc actgcacgcc gtaggtcagg gtggtcacga gggtgggcca
      121 gggcacgggc agcttgccgg tggtgcagat gaacttcagg gtcagcttgc cgtaggtggc
      181 atcgccctcg ccctcgccgg acacgctgaa cttgtggccg tttacgtcgc cgtccagctc
      241 gaccaggatg ggcaccaccc cggtgaacag ctcctcgccc ttgctcactg ggccaggatt
      301 ctcctcgacg tcaccgcatg ttagcagact tcctctgccc tctccactgc cgaattcctt
      361 tttctttttt gcctggccgg cctttttcgt ggccgccggc cttttgtcgc ctcccagctg
      421 agacaggtcg atccgtgtct cgtacaggcc ggtgatgctc tggtggatca gggtggcgtc
      481 cagcacctct ttggtgctgg tgtacctctt ccggtcgatg gtggtgtcaa agtacttgaa
      541 ggcggcaggg gctcccagat tggtcagggt aaacaggtgg atgatattct cggcctgctc
      601 tctgatgggc ttatcccggt gcttgttgta ggcggacagc actttgtcca gattagcgtc
      661 ggccaggatc actctcttgg agaactcgct gatctgctcg atgatctcgt ccaggtagtg
      721 cttgtgctgt tccacaaaca gctgtttctg ctcattatcc tcgggggagc ccttcagctt
      781 ctcatagtgg ctggccaggt acaggaagtt cacatatttg gagggcaggg ccagttcgtt
      841 tcccttctgc agttcgccgg cagaggccag cattctcttc cggccgtttt ccagctcgaa
      901 cagggagtac ttaggcagct tgatgatcag gtcctttttc acttctttgt agcccttggc
      961 ttccagaaag tcgatgggat tcttctcgaa gctgcttctt tccatgatgg tgatccccag
     1021 cagctctttc acactcttca gtttcttgga cttgcccttt tccactttgg ccaccaccag
     1081 cacagaatag gccacggtgg ggctgtcgaa gccgccgtac ttcttagggt cccagtcctt
     1141 ctttctggcg a
//
