LOCUS       sequence-96694-e         571 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Co-expresses human optimized S. pyogenes dCas9 fused to two copies 
            of VP64 and GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 571)
  AUTHORS   Kabadi AM, Ousterout DG, Hilton IB, Gersbach CA
  TITLE     Multiplex CRISPR/Cas9-based genome engineering from a single 
            lentiviral vector.
  JOURNAL   Nucleic Acids Res. 2014 Aug 13. pii: gku749.
  PUBMED    25122746
REFERENCE   2  (bases 1 to 571)
  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: "Nucleic 
            Acids Res. 2014 Aug 13. pii: gku749."
COMMENT     Sequence Label: pLV hUbC-VP64 dCas9 VP64-T2A-GFP
FEATURES             Location/Qualifiers
     source          1..571
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      118..127
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             127..192
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             208..354
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation 
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label=VP64
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDM"
     CDS             361..381
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 tgtaatcatt tgggtcaata tgtaattttc agtgttagac tagtaaattg tccgctaaat
       61 tctggccgtt tttggctttt ttgttagacg aagcttgggc tgcaggtcga ctctagagcc
      121 accatggact acaaagacca tgacggtgat tataaagatc atgacatcga ttacaaggat
      181 gacgatgaca agcacggtcc gcgggctgac gcattggacg attttgatct ggatatgctg
      241 ggaagtgacg ccctcgatga ttttgacctt gacatgcttg gttcggatgc ccttgatgac
      301 tttgacctcg acatgctcgg cagtgacgcc cttgatgatt tcgacctgga catggttaac
      361 cccaagaaga agaggaaggt gggccgcgga atggacaaga agtactccat tgggctcgcc
      421 atcggcacaa acagcgtcgg ctgggccgtc attacggacg agtacaaggt gccgagcaaa
      481 aaattcaaag ttctgggcaa taccgatcgc cacagcataa agaagaacct cattggcgcc
      541 ctcctgttcg actccgggga aaccgccgaa g
//
