LOCUS       sequence-64963-e         628 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 628)
  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 628)
  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..628
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(330..350)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(357..422)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     regulatory      422..431
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 tcttccagtc tgtggaagaa gctgtcgtcc accttggcca tctcgttgct gaagatctct
       61 tgcagatagc agatccggtt cttccgtctg gtgtatcttc ttctggcggt tctcttcagc
      121 cgggtggcct cggctgtttc gccgctgtcg aacagcaggg ctccgatcag gttcttcttg
      181 atgctgtgcc ggtcggtgtt gcccagcacc ttgaatttct tgctgggcac cttgtactcg
      241 tcggtgatca cggcccagcc cacagagttg gtgccgatgt ccaggccgat gctgtacttc
      301 ttgtcggctg ctgggactcc gtggataccg accttccgct tcttctttgg ggccatctta
      361 tcgtcatcgt ctttgtaatc aatatcatga tccttgtagt ctccgtcgtg gtccttatag
      421 tccatggtgg caccggtcca acctgaaaaa aagtgatttc aggcaggtgc tccaggtaat
      481 taaacattaa taccccacca accaaccatc ccttaaaccc ttacctcttg ctcagctaat
      541 tacagcccgg aggagaaggg ccgtcccgcc cgctcacctg tgggagtaac gcggtcagtc
      601 agagccgggg cgggcggcgc gaggcggc
//
