LOCUS       sequence-95560-e         529 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 529)
  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 529)
  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-1x6
FEATURES             Location/Qualifiers
     source          1..529
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(325..345)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(352..417)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     regulatory      417..426
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 cagtctgtgg aagaagctgt cgtccacctt ggccatctcg ttgctgaaga tctcttgcag
       61 atagcagatc cggttcttcc gtctggtgta tcttcttctg gcggttctct tcagccgggt
      121 ggcctcggct gtttcgccgc tgtcgaacag cagggctccg atcaggttct tcttgatgct
      181 gtgccggtcg gtgttgccca gcaccttgaa tttcttgctg ggcaccttgt actcgtcggt
      241 gatcacggcc cagcccacag agttggtgcc gatggccagg ccgatgctgt acttcttgtc
      301 ggctgctggg actccgtgga taccgacctt ccgcttcttc tttggggcca tcttatcgtc
      361 atcgtctttg taatcaatat catgatcctt gtagtctccg tcgtggtcct tatagtccat
      421 ggtggcaccg gtccaacctg aaaaaaagtg atttcaggca ggtgctccag gtaattaaac
      481 attaataccc caccaaccaa ccatccctta aacccttacc tcttgctca
//
