LOCUS       sequence-99326-e         596 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 596)
  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 596)
  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-1x2
FEATURES             Location/Qualifiers
     source          1..596
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(311..331)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(338..403)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     regulatory      403..412
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 agctgtcgtc caccttggcc atctcgttgc tgaagatctc ttgcagatag cagatccggt
       61 tcttccgtct ggtgtatctt cttctggcgg ttctcttcag ccgggtggcc tcggctgttt
      121 cgccgctgtc gaacagcagg gctccgatca ggttcttctt gatgctgtgc cggtcggtgt
      181 tgcccagcac cttgaatttc ttgctgggca ccttgtactc gtcggtgatc acggcccagc
      241 ccacagagtt ggtgccgatg gccaggccga tgctgtactt cttgtcggct gctgggactc
      301 cgtggatacc gaccttccgc ttcttctttg gggccatctt atcgtcatcg tctttgtaat
      361 caatatcatg atccttgtag tctccgtcgt ggtccttata gtccatggtg gcaccggtcc
      421 aacctgaaaa aaagtgattt caggcaggtg ctccaggtaa ttaaacatta ataccccacc
      481 aaccaaccat cccttaaacc cttacctctt gctcagctaa ttacagcccg gaggagaagg
      541 gccgtcccgc ccgctcacct gtgggagtaa cgcggtcagt cagagccggg gcgggc
//
