LOCUS       sequence-73275-e        1002 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  NOTE: A new version of this plasmid is now available. See Addgene 
            plasmid 62987.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1002)
  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 1002)
  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-Puro (PX462)
FEATURES             Location/Qualifiers
     source          1..1002
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      59..68
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             68..133
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             140..160
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 taatgtttaa ttacctggag cacctgcctg aaatcacttt ttttcaggtt ggaccggtgc
       61 caccatggac tataaggacc acgacggaga ctacaaggat catgatattg attacaaaga
      121 cgatgacgat aagatggccc caaagaagaa gcggaaggtc ggtatccacg gagtcccagc
      181 agccgacaag aagtacagca tcggcctggc catcggcacc aactctgtgg gctgggccgt
      241 gatcaccgac gagtacaagg tgcccagcaa gaaattcaag gtgctgggca acaccgaccg
      301 gcacagcatc aagaagaacc tgatcggagc cctgctgttc gacagcggcg aaacagccga
      361 ggccacccgg ctgaagagaa ccgccagaag aagatacacc agacggaaga accggatctg
      421 ctatctgcaa gagatcttca gcaacgagat ggccaaggtg gacgacagct tcttccacag
      481 actggaagag tccttcctgg tggaagagga taagaagcac gagcggcacc ccatcttcgg
      541 caacatcgtg gacgaggtgg cctaccacga gaagtacccc accatctacc acctgagaaa
      601 gaaactggtg gacagcaccg acaaggccga cctgcggctg atctatctgg ccctggccca
      661 catgatcaag ttccggggcc acttcctgat cgagggcgac ctgaaccccg acaacagcga
      721 cgtggacaag ctgttcatcc agctggtgca gacctacaac cagctgttcg aggaaaaccc
      781 catcaacgcc agcggcgtgg acgccaaggc catcctgtct gccagactga gcaagagcag
      841 acggctggaa aatctgatcg cccagctgcc cggcgagaag aagaatggcc tgttcggcaa
      901 cctgattgcc ctgagcctgg gcctgacccc caacttcaag agcaacttcg acctggccga
      961 ggatgccaaa ctgcagctga gcaaggacac ctacgacgac ga
//
