LOCUS       sequence-122239-         635 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  expresses VP64-dCas9-VP64, same plasmid as Addgene 59791 but with 
            LoxP sites removed.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 635)
  AUTHORS   Gersbach C
  TITLE     Multiplex CRISPR/Cas9 Genome Engineering for Directing Myogenic 
            Cellular Differentiation in Primary Human Cells
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 635)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pLV hUbC- VP64-dCas9-VP64-T2A- GFP(No LoxP sites)
FEATURES             Location/Qualifiers
     source          1..635
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      138..147
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             147..212
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             228..374
                     /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             381..401
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 tttttaggca ccttttgaaa tgtaatcatt tgggtcaata tgtaattttc agtgttagac
       61 tagtaaattg tccgctaaat tctggccgtt tttggctttt ttgttagacg aagcttgggc
      121 tgcaggtcga ctctagagcc accatggact acaaagacca tgacggtgat tataaagatc
      181 atgacatcga ttacaaggat gacgatgaca agcacggtcc gcgggctgac gcattggacg
      241 attttgatct ggatatgctg ggaagtgacg ccctcgatga ttttgacctt gacatgcttg
      301 gttcggatgc ccttgatgac tttgacctcg acatgctcgg cagtgacgcc cttgatgatt
      361 tcgacctgga catggttaac cccaagaaga agaggaaggt gggccgcgga atggacaaga
      421 agtactccat tgggctcgcc atcggcacaa acagcgtcgg ctgggccgtc attacggacg
      481 agtacaaggt gccgagcaaa aaattcaaag ttctgggcaa taccgatcgc cacagcataa
      541 agaagaacct cattggcgcc ctcctgttcg actccgggga aaccgccgaa gccacgcggc
      601 tcaaaagaac agcacggcgc agatataccc gcaga
//
