LOCUS       sequence-75302-e         951 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  A human codon-optimized SpCas9 and chimeric guide RNA expression 
            plasmid..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 951)
  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 951)
  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: pX330-U6-Chimeric_BB-CBh-hSpCas9
FEATURES             Location/Qualifiers
     source          1..951
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        122..370
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     122..142
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     293..312
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        389..464
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        561..846
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
       61 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
      121 tgagggccta tttcccatga ttccttcata tttgcatata cgatacaagg ctgttagaga
      181 gataattgga attaatttga ctgtaaacac aaagatatta gtacaaaata cgtgacgtag
      241 aaagtaataa tttcttgggt agtttgcagt tttaaaatta tgttttaaaa tggactatca
      301 tatgcttacc gtaacttgaa agtatttcga tttcttggct ttatatatct tgtggaaagg
      361 acgaaacacc gggtcttcga gaagacctgt tttagagcta gaaatagcaa gttaaaataa
      421 ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg gtgctttttt gttttagagc
      481 tagaaatagc aagttaaaat aaggctagtc cgtttttagc gcgtgcgcca attctgcaga
      541 caaatggctc tagaggtacc cgttacataa cttacggtaa atggcccgcc tggctgaccg
      601 cccaacgacc cccgcccatt gacgtcaata gtaacgccaa tagggacttt ccattgacgt
      661 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
      721 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttgtgcccag
      781 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
      841 accatggtcg aggtgagccc cacgttctgc ttcactctcc ccatctcccc cccctcccca
      901 cccccaattt tgtatttatt tattttttaa ttattttgtg cagcgatggg g
//
