LOCUS       sequence-97942-e         778 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  A human codon-optimized SpCas9 nickase and chimeric guide RNA 
            expression plasmid..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 778)
  AUTHORS   Cong L, Ran FA, Cox D, Lin S, Barretto R, Habib N, Hsu PD, Wu X, 
            Jiang W, Marraffini LA, Zhang F
  TITLE     Multiplex Genome Engineering Using CRISPR/Cas Systems.
  JOURNAL   Science. 2013 Jan 3.
  PUBMED    23287718
REFERENCE   2  (bases 1 to 778)
  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: "Science. 
            2013 Jan 3."
COMMENT     Sequence Label: pX335-U6-Chimeric_BB-CBh-hSpCas9n(D10A)
FEATURES             Location/Qualifiers
     source          1..778
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(315..335)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(339..365)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     intron          386..613
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
ORIGIN
        1 tcttccagtc tgtggaagaa gctgtcgtcc accttggcca tctcgttgct gaagatctct
       61 tgcagatagc agatccggtt cttccgtctg gtgtatcttc ttctggcggt tctcttcagc
      121 cgggtggcct cggctgtttc gccgctgtcg aacagcaggg ctccgatcag gttcttcttg
      181 atgctgtgcc ggtcggtgtt gcccagcacc ttgaatttct tgctgggcac cttgtactcg
      241 tcggtgatca cggcccagcc cacagagttg gtgccgatgg ccaggccgat gctgtacttc
      301 ttgtcggacg cttcgacctt gcgctttttc ttcggcgaag cgtaatctgg aacatcgtat
      361 gggtacatgg tggcaccggt ccaacctgaa aaaaagtgat ttcaggcagg tgctccaggt
      421 aattaaacat taatacccca ccaaccaacc atcccttaaa cccttacctc ttgctcagct
      481 aattacagcc cggaggagaa gggccgtccc gcccgctcac ctgtgggagt aacgcggtca
      541 gtcagagccg gggcgggcgg cgcgaggcgg cggcggagcg gggcacgggg cgaaggcagc
      601 gcgcagcgac tcccgcccgc cgcgcgcttc gctttttata gggccgccgc cgccgccgcc
      661 tcgccataaa aggaaacttt cggagcgcgc cgctctgatt ggctgccgcc gcacctctcc
      721 gcctcgcccc gccccgcccc tcgccccgcc ccgccccgcc tggcgcgcgc cccccccc
//
