LOCUS       sequence-63665-e         894 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 894)
  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 894)
  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..894
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    59..92
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        153..652
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(188..211)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     585..604
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     complement(692..711)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
ORIGIN
        1 tctcggcatg gacgagctgt acaagtaaga attccgatca tattcaataa cccttaatat
       61 aacttcgtat aatgtatgct atacgaagtt attaggtctg aagaggagtt tacgtccagc
      121 caagcttagg atctcgacct cgaaattcta ccgggtaggg gaggcgcttt tcccaaggca
      181 gtctggagca tgcgctttag cagccccgct gggcacttgg cgctacacaa gtggcctctg
      241 gcctcgcaca cattccacat ccaccggtag gcgccaaccg gctccgttct ttggtggccc
      301 cttcgcgcca ccttctactc ctcccctagt caggaagttc ccccccgccc cgcagctcgc
      361 gtcgtgcagg acgtgacaaa tggaagtagc acgtctcact agtctcgtgc agatggacag
      421 caccgctgag caatggaagc gggtaggcct ttggggcagc ggccaatagc agctttgctc
      481 cttcgctttc tgggctcaga ggctgggaag gggtgggtcc gggggcgggc tcaggggcgg
      541 gctcaggggc ggggcgggcg cccgaaggtc ctccggaggc ccggcattct gcacgcttca
      601 aaagcgcacg tctgccgcgc tgttctcctc ttcctcatct ccgggccttt cgacctgcat
      661 ccatctagat ctcgagcagc tgaagcttac catgaccgag tacaagccca cggtgcgcct
      721 cgccacccgc gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta
      781 ccccgccacg cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca
      841 agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cgga
//
