LOCUS       sequence-95335-e         919 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  This plasmid separately encodes a human codon-optimized SpCas9, a 
            tracrRNA and customizable crRNA..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 919)
  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 919)
  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: 
            pX260-U6-DR-BB-DR-Cbh-NLS-hSpCas9-NLS-H1-shorttracr-PGK-puro
FEATURES             Location/Qualifiers
     source          1..919
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        127..375
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     127..147
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     298..317
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     repeat_region   378..413
                     /label=DR
                     /note="direct repeat for the Streptococcus pyogenes 
                     CRISPR/Cas system"
     repeat_region   430..465
                     /label=DR
                     /note="direct repeat for the Streptococcus pyogenes 
                     CRISPR/Cas system"
     enhancer        516..801
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
       61 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
      121 acatgtgagg gcctatttcc catgattcct tcatatttgc atatacgata caaggctgtt
      181 agagagataa ttggaattaa tttgactgta aacacaaaga tattagtaca aaatacgtga
      241 cgtagaaagt aataatttct tgggtagttt gcagttttaa aattatgttt taaaatggac
      301 tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct tggctttata tatcttgtgg
      361 aaaggacgaa acaccgggtt ttagagctat gctgttttga atggtcccaa aacgggtctt
      421 cgagaagacg ttttagagct atgctgtttt gaatggtccc aaaacttttt ctagcgcgtg
      481 cgccaattct gcagacaaat ggctctagag gtacccgtta cataacttac ggtaaatggc
      541 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caatagtaac gccaataggg
      601 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
      661 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
      721 tggcattgtg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
      781 ttagtcatcg ctattaccat ggtcgaggtg agccccacgt tctgcttcac tctccccatc
      841 tcccccccct ccccaccccc aattttgtat ttatttattt tttaattatt ttgtgcagcg
      901 atgggggcgg ggggggggg
//
