LOCUS       sequence-65408-e        1009 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Dual expression plasmid of human codon-optimized SpCas9 and a gRNA 
            to the human Emx1 locus, can be used to test SpCas9 cleavage in cell
            lines of choice..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1009)
  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 1009)
  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: 
            pX261-U6-DR-hEmx1-DR-Cbh-NLS-hSpCas9-NLS-H1-shorttracr-PGK-puro
FEATURES             Location/Qualifiers
     source          1..1009
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        complement(19..518)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(67..86)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     460..483
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     protein_bind    complement(579..612)
                     /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(688..709)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_RNA        complement(783..861)
                     /label=tracrRNA
                     /note="trans-activating CRISPR RNA for the Streptococcus 
                     pyogenes CRISPR/Cas9 system"
     promoter        complement(872..962)
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     primer_bind     complement(932..951)
                     /label=H1
                     /note="Human H1 promoter, forward primer"
ORIGIN
        1 atctagatgg atgcaggtcg aaaggcccgg agatgaggaa gaggagaaca gcgcggcaga
       61 cgtgcgcttt tgaagcgtgc agaatgccgg gcctccggag gaccttcggg cgcccgcccc
      121 gcccctgagc ccgcccctga gcccgccccc ggacccaccc cttcccagcc tctgagccca
      181 gaaagcgaag gagcaaagct gctattggcc gctgccccaa aggcctaccc gcttccattg
      241 ctcagcggtg ctgtccatct gcacgagact agtgagacgt gctacttcca tttgtcacgt
      301 cctgcacgac gcgagctgcg gggcgggggg gaacttcctg actaggggag gagtagaagg
      361 tggcgcgaag gggccaccaa agaacggagc cggttggcgc ctaccggtgg atgtggaatg
      421 tgtgcgaggc cagaggccac ttgtgtagcg ccaagtgccc agcggggctg ctaaagcgca
      481 tgctccagac tgccttggga aaagcgcctc ccctacccgg tagaatttcg aggtcgagat
      541 cctaagcttg gctggacgta aactcctctt cagacctaat aacttcgtat agcatacatt
      601 atacgaagtt atattaaggg ttattgaata tgatcggaat tcttacttgt acagctcgtc
      661 catgccgaga gtgatcccgg cggcggtcac gaactccagc aggaccatgt gatcgcgctt
      721 ctcgttgggg tctttgctca gggcggactg ggtgctcagg tagtggttgt cggcgccaaa
      781 aagcaccgac tcggtgccac tttttcaagt tgataacgga ctagccttat tttaacttgc
      841 tatgctgttt tgaatggttc caactagatc tgtggtctca tacagaactt ataagattcc
      901 caaatccaaa gacatttcac gtttatggtg atttcccaga acacatagcg acatgcaaat
      961 attgcagggc gccactcccc tgtccctcac agccatcttc ctgccaggg
//
