LOCUS       sequence-65403-e        1025 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 1025)
  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 1025)
  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..1025
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      55..64
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             64..129
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             136..156
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 gtttaattac ctggagcacc tgcctgaaat cacttttttt caggttggac cggtgccacc
       61 atggactata aggaccacga cggagactac aaggatcatg atattgatta caaagacgat
      121 gacgataaga tggccccaaa gaagaagcgg aaggtcggta tccacggagt cccagcagcc
      181 gacaagaagt acagcatcgg cctggacatc ggcaccaact ctgtgggctg ggccgtgatc
      241 accgacgagt acaaggtgcc cagcaagaaa ttcaaggtgc tgggcaacac cgaccggcac
      301 agcatcaaga agaacctgat cggagccctg ctgttcgaca gcggcgaaac agccgaggcc
      361 acccggctga agagaaccgc cagaagaaga tacaccagac ggaagaaccg gatctgctat
      421 ctgcaagaga tcttcagcaa cgagatggcc aaggtggacg acagcttctt ccacagactg
      481 gaagagtcct tcctggtgga agaggataag aagcacgagc ggcaccccat cttcggcaac
      541 atcgtggacg aggtggccta ccacgagaag taccccacca tctaccacct gagaaagaaa
      601 ctggtggaca gcaccgacaa ggccgacctg cggctgatct atctggccct ggcccacatg
      661 atcaagttcc ggggccactt cctgatcgag ggcgacctga accccgacaa cagcgacgtg
      721 gacaagctgt tcatccagct ggtgcagacc tacaaccagc tgttcgagga aaaccccatc
      781 aacgccagcg gcgtggacgc caaggccatc ctgtctgcca gactgagcaa gagcagacgg
      841 ctggaaaatc tgatcgccca gctgcccggc gagaagaaga atggcctgtt cggaaacctg
      901 attgccctga gcctgggcct gacccccaac ttcaagagca acttcgacct ggccgaggat
      961 gccaaactgc agctgagcaa ggacacctac gacgacgacc tggacaacct gctggcccag
     1021 atcgg
//
