LOCUS       sequence-70335-e         969 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 969)
  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 969)
  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..969
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      60..69
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             69..134
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             141..161
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 ttaatgttta attacctgga gcacctgcct gaaatcactt tttttcaggt tggaccggtg
       61 ccaccatgga ctataaggac cacgacggag actacaagga tcatgatatt gattacaaag
      121 acgatgacga taagatggcc ccaaagaaga agcggaaggt cggtatccac ggagtcccag
      181 cagccgacaa gaagtacagc atcggcctgg acatcggcac caactctgtg ggctgggccg
      241 tgatcaccga cgagtacaag gtgcccagca agaaattcaa ggtgctgggc aacaccgacc
      301 ggcacagcat caagaagaac ctgatcggag ccctgctgtt cgacagcggc gaaacagccg
      361 aggccacccg gctgaagaga accgccagaa gaagatacac cagacggaag aaccggatct
      421 gctatctgca agagatcttc agcaacgaga tggccaaggt ggacgacagc ttcttccaca
      481 gactggaaga gtccttcctg gtggaagagg ataagaagca cgagcggcac cccatcttcg
      541 gcaacatcgt ggacgaggtg gcctaccacg agaagtaccc caccatctac cacctgagaa
      601 agaaactggt ggacagcacc gacaaggccg acctgcggct gatctatctg gccctggccc
      661 acatgatcaa gttccggggc cacttcctga tcgagggcga cctgaacccc gacaacagcg
      721 acgtggacaa gctgttcatc cagctggtgc agacctacaa ccagctgttc gaggaaaacc
      781 ccatcaacgc cagcggcgtg gacgccaagg ccatcctgtc tgccagactg agcaagagca
      841 gacggctgga aaatctgatc gcccagctgc ccggcgagaa gaagaatggc ctgttcggaa
      901 acctgattgc cctgagcctg ggcctgaccc ccaacttcaa gagcaacttc gacctggccg
      961 aggatgcca
//
