LOCUS       sequence-93045-e         573 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses Cas9 nickase and gRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 573)
  AUTHORS   Sakuma T, Nishikawa A, Kume S, Chayama K, Yamamoto T
  TITLE     Multiplex genome engineering in human cells using all-in-one 
            CRISPR/Cas9 vector system.
  JOURNAL   Sci Rep. 2014 Jun 23;4:5400. doi: 10.1038/srep05400.
  PUBMED    24954249
REFERENCE   2  (bases 1 to 573)
  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"; journalName: "Sci Rep."; 
            date: "2014-06-23"; volume: "4:5400. doi"; pages: " 
            10.1038/srep05400"
COMMENT     Sequence Label: pX330A_D10A-1x3
FEATURES             Location/Qualifiers
     source          1..573
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      28..37
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             37..102
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             109..129
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 aatcactttt tttcaggttg gaccggtgcc accatggact ataaggacca cgacggagac
       61 tacaaggatc atgatattga ttacaaagac gatgacgata agatggcccc aaagaagaag
      121 cggaaggtcg gtatccacgg agtcccagca gccgacaaga agtacagcat cggcctggcc
      181 atcggcacca actctgtggg ctgggccgtg atcaccgacg agtacaaggt gcccagcaag
      241 aaattcaagg tgctgggcaa caccgaccgg cacagcatca agaagaacct gatcggagcc
      301 ctgctgttcg acagcggcga aacagccgag gccacccggc tgaagagaac cgccagaaga
      361 agatacacca gacggaagaa ccggatctgc tatctgcaag agatcttcag caacgagatg
      421 gccaaggtgg acgacagctt cttccacaga ctggaagagt ccttcctggt ggaagaggat
      481 aagaagcacg agcggcaccc catcttcggc aacatcgtgg acgaggtggc ctaccacgag
      541 aagtacccca ccatctacca cctgagaaag aaa
//
