LOCUS       sequence-64844-e        1000 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 1000)
  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 1000)
  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..1000
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        complement(25..524)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(73..92)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     466..489
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     protein_bind    complement(585..618)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(694..715)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_RNA        complement(789..867)
                     /label=tracrRNA
                     /note="trans-activating CRISPR RNA for the Streptococcus 
                     pyogenes CRISPR/Cas9 system"
     promoter        complement(878..968)
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     primer_bind     complement(938..957)
                     /label=H1
                     /note="Human H1 promoter, forward primer"
ORIGIN
        1 ctcgagatct agatggatgc aggtcgaaag gcccggagat gaggaagagg agaacagcgc
       61 ggcagacgtg cgcttttgaa gcgtgcagaa tgccgggcct ccggaggacc ttcgggcgcc
      121 cgccccgccc ctgagcccgc ccctgagccc gcccccggac ccaccccttc ccagcctctg
      181 agcccagaaa gcgaaggagc aaagctgcta ttggccgctg ccccaaaggc ctacccgctt
      241 ccattgctca gcggtgctgt ccatctgcac gagactagtg agacgtgcta cttccatttg
      301 tcacgtcctg cacgacgcga gctgcggggc gggggggaac ttcctgacta ggggaggagt
      361 agaaggtggc gcgaaggggc caccaaagaa cggagccggt tggcgcctac cggtggatgt
      421 ggaatgtgtg cgaggccaga ggccacttgt gtagcgccaa gtgcccagcg gggctgctaa
      481 agcgcatgct ccagactgcc ttgggaaaag cgcctcccct acccggtaga atttcgaggt
      541 cgagatccta agcttggctg gacgtaaact cctcttcaga cctaataact tcgtatagca
      601 tacattatac gaagttatat taagggttat tgaatatgat cggaattctt acttgtacag
      661 ctcgtccatg ccgagagtga tcccggcggc ggtcacgaac tccagcagga ccatgtgatc
      721 gcgcttctcg ttggggtctt tgctcagggc ggactgggtg ctcaggtagt ggttgtcggc
      781 gccaaaaagc accgactcgg tgccactttt tcaagttgat aacggactag ccttatttta
      841 acttgctatg ctgttttgaa tggttccaac tagatctgtg gtctcataca gaacttataa
      901 gattcccaaa tccaaagaca tttcacgttt atggtgattt cccagaacac atagcgacat
      961 gcaaatattg cagggcgcca ctcccctgtc cctcacagcc
//
