LOCUS       sequence-78358-e        1129 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 1129)
  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 1129)
  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..1129
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      60..69
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             69..134
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             141..161
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 ttaatgttta attacctgga gcacctgcct gaaatcactt tttttcaggt tggaccggtg
       61 ccaccatgga ctataaggac cacgacggag actacaagga tcatgatatt gattacaaag
      121 acgatgacga taagatggcc ccaaagaaga agcggaaggt cggtatccac ggagtcccag
      181 cagccgacaa gaagtacagc atcggcctgg ccatcggcac caactctgtg ggctgggccg
      241 tgatcaccga cgagtacaag gtgcccagca agaaattcaa ggtgctgggc aacaccgacc
      301 ggcacagcat caagaagaac ctgatcggag ccctgctgtt cgacagcggc gaaacagccg
      361 aggccacccg gctgaagaga accgccagaa gaagatacac cagacggaag aaccggatct
      421 gctatctgca agagatcttc agcaacgaga tggccaaggt ggacgacagc ttcttccaca
      481 gactggaaga gtccttcctg gtggaagagg ataagaagca cgagcggcac cccatcttcg
      541 gcaacatcgt ggacgaggtg gcctaccacg agaagtaccc caccatctac cacctgagaa
      601 agaaactggt ggacagcacc gacaaggccg acctgcggct gatctatctg gccctggccc
      661 acatgatcaa gttccggggc cacttcctga tcgagggcga cctgaacccc gacaacagcg
      721 acgtggacaa gctgttcatc cagctggtgc agacctacaa ccagctgttc gaggaaaacc
      781 ccatcaacgc cagcggcgtg gacgccaagg ccatcctgtc tgccagactg agcaagagca
      841 gacggctgga aaatctgatc gcccagctgc ccggcgagaa gaagaatggc ctgttcggca
      901 acctgattgc cctgagcctg ggcctgaccc ccaacttcaa gagcaacttc gacctggccg
      961 aggatgccaa actgcagctg agcaaggaca cctacgacga cgacctggac aacctgctgg
     1021 cccagatcgg cgaccagtac gccgacctgt ttctggccgc caagaacctg tccgacgcca
     1081 tcctgctgag cgacatcctg agagtgaaca ccgagatcac caaggcccc
//
