LOCUS       sequence-63088-e         727 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  This plasmid separately encodes a human codon-optimized SpCas9 
            nickase, a tracrRNA and customizable crRNA..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 727)
  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 727)
  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: 
            pX334-U6-DR-BB-DR-Cbh-NLS-hSpCas9n(D10A)-NLS-H1-shorttracr-PGK-puro
FEATURES             Location/Qualifiers
     source          1..727
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(313..333)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(337..363)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     intron          384..611
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
ORIGIN
        1 ttccagtctg tggaagaagc tgtcgtccac cttggccatc tcgttgctga agatctcttg
       61 cagatagcag atccggttct tccgtctggt gtatcttctt ctggcggttc tcttcagccg
      121 ggtggcctcg gctgtttcgc cgctgtcgaa cagcagggct ccgatcaggt tcttcttgat
      181 gctgtgccgg tcggtgttgc ccagcacctt gaatttcttg ctgggcacct tgtactcgtc
      241 ggtgatcacg gcccagccca cagagttggt gccgatggcc aggccgatgc tgtacttctt
      301 gtcggacgct tcgaccttgc gctttttctt cggcgaagcg taatctggaa catcgtatgg
      361 gtacatggtg gcaccggtcc aacctgaaaa aaagtgattt caggcaggtg ctccaggtaa
      421 ttaaacatta ataccccacc aaccaaccat cccttaaacc cttacctctt gctcagctaa
      481 ttacagcccg gaggagaagg gccgtcccgc ccgctcacct gtgggagtaa cgcggtcagt
      541 cagagccggg gcgggcggcg cgaggcggcg gcggagcggg gcacggggcg aaggcagcgc
      601 gcagcgactc ccgcccgccg cgcgcttcgc tttttatagg gccgccgccg ccgccgcctc
      661 gccataaaag gaaactttcg gagcgcgccg ctctgattgg ctgccgccgc acctctccgc
      721 ctcgccc
//
