LOCUS       sequence-99195-e        1124 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  NOTE: A new version of this plasmid is now available. See Addgene 
            plasmid 62988..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1124)
  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 1124)
  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: pSpCas9(BB)-2A-Puro (PX459)
FEATURES             Location/Qualifiers
     source          1..1124
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      57..66
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             66..131
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             138..158
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 atgtttaatt acctggagca cctgcctgaa atcacttttt ttcaggttgg accggtgcca
       61 ccatggacta taaggaccac gacggagact acaaggatca tgatattgat tacaaagacg
      121 atgacgataa gatggcccca aagaagaagc ggaaggtcgg tatccacgga gtcccagcag
      181 ccgacaagaa gtacagcatc ggcctggaca tcggcaccaa ctctgtgggc tgggccgtga
      241 tcaccgacga gtacaaggtg cccagcaaga aattcaaggt gctgggcaac accgaccggc
      301 acagcatcaa gaagaacctg atcggagccc tgctgttcga cagcggcgaa acagccgagg
      361 ccacccggct gaagagaacc gccagaagaa gatacaccag acggaagaac cggatctgct
      421 atctgcaaga gatcttcagc aacgagatgg ccaaggtgga cgacagcttc ttccacagac
      481 tggaagagtc cttcctggtg gaagaggata agaagcacga gcggcacccc atcttcggca
      541 acatcgtgga cgaggtggcc taccacgaga agtaccccac catctaccac ctgagaaaga
      601 aactggtgga cagcaccgac aaggccgacc tgcggctgat ctatctggcc ctggcccaca
      661 tgatcaagtt ccggggccac ttcctgatcg agggcgacct gaaccccgac aacagcgacg
      721 tggacaagct gttcatccag ctggtgcaga cctacaacca gctgttcgag gaaaacccca
      781 tcaacgccag cggcgtggac gccaaggcca tcctgtctgc cagactgagc aagagcagac
      841 ggctggaaaa tctgatcgcc cagctgcccg gcgagaagaa gaatggcctg ttcggaaacc
      901 tgattgccct gagcctgggc ctgaccccca acttcaagag caacttcgac ctggccgagg
      961 atgccaaact gcagctgagc aaggacacct acgacgacga cctggacaac ctgctggccc
     1021 agatcggcga ccagtacgcc gacctgtttc tggccgccaa gaacctgtcc gacgccatcc
     1081 tgctgagcga catcctgaga gtgaacaccg agatcaccaa ggcc
//
