LOCUS       sequence-95571-e         513 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 513)
  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 513)
  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-1x7
FEATURES             Location/Qualifiers
     source          1..513
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(309..329)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(336..401)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     regulatory      401..410
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 ctgtcgtcca ccttggccat ctcgttgctg aagatctctt gcagatagca gatccggttc
       61 ttccgtctgg tgtatcttct tctggcggtt ctcttcagcc gggtggcctc ggctgtttcg
      121 ccgctgtcga acagcagggc tccgatcagg ttcttcttga tgctgtgccg gtcggtgttg
      181 cccagcacct tgaatttctt gctgggcacc ttgtactcgt cggtgatcac ggcccagccc
      241 acagagttgg tgccgatggc caggccgatg ctgtacttct tgtcggctgc tgggactccg
      301 tggataccga ccttccgctt cttctttggg gccatcttat cgtcatcgtc tttgtaatca
      361 atatcatgat ccttgtagtc tccgtcgtgg tccttatagt ccatggtggc accggtccaa
      421 cctgaaaaaa agtgatttca ggcaggtgct ccaggtaatt aaacattaat accccaccaa
      481 ccaaccatcc cttaaaccct tacctcttgc tca
//
