LOCUS       sequence-63168-e         844 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 844)
  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 844)
  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..844
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        124..372
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     124..144
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     295..314
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     repeat_region   375..410
                     /label=DR
                     /note="direct repeat for the Streptococcus pyogenes 
                     CRISPR/Cas system"
     repeat_region   427..462
                     /label=DR
                     /note="direct repeat for the Streptococcus pyogenes 
                     CRISPR/Cas system"
     enhancer        513..798
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
       61 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
      121 tgtgagggcc tatttcccat gattccttca tatttgcata tacgatacaa ggctgttaga
      181 gagataattg gaattaattt gactgtaaac acaaagatat tagtacaaaa tacgtgacgt
      241 agaaagtaat aatttcttgg gtagtttgca gttttaaaat tatgttttaa aatggactat
      301 catatgctta ccgtaacttg aaagtatttc gatttcttgg ctttatatat cttgtggaaa
      361 ggacgaaaca ccgggtttta gagctatgct gttttgaatg gtcccaaaac gggtcttcga
      421 gaagacgttt tagagctatg ctgttttgaa tggtcccaaa actttttcta gcgcgtgcgc
      481 caattctgca gacaaatggc tctagaggta cccgttacat aacttacggt aaatggcccg
      541 cctggctgac cgcccaacga cccccgccca ttgacgtcaa tagtaacgcc aatagggact
      601 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
      661 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
      721 cattgtgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
      781 gtcatcgcta ttaccatggt cgaggtgagc cccacgttct gcttcactct ccccatctcc
      841 cccc
//
