LOCUS       sequence-154069-         852 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses MCP:VP64 under actin promoter for ""Scaffold"" CRISPRa in 
            Drosophila cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 852)
  AUTHORS   Chavez A, Tuttle M, Pruitt BW, Ewen-Campen B, Chari R, Ter-Ovanesyan
            D, Haque SJ, Cecchi RJ, Kowal EJ, Buchthal J, Housden BE, Perrimon 
            N, Collins JJ, Church G
  TITLE     Comparison of Cas9 activators in multiple species.
  JOURNAL   Nat Methods. 2016 May 23. doi: 10.1038/nmeth.3871.
  PUBMED    27214048
REFERENCE   2  (bases 1 to 852)
  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: "Nat 
            Methods. 2016 May 23. doi: 10.1038/nmeth.3871."
COMMENT     Sequence Label: pAct:MCP-VP64
FEATURES             Location/Qualifiers
     source          1..852
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      28..37
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             40..60
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             70..93
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             475..624
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation 
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label=VP64
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
     protein_bind    complement(641..665)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      673..682
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(724..745)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
ORIGIN
        1 agtttgtaca aaaaagcagg ctacaaagcc accatgggac ctaagaaaaa gaggaaggtg
       61 gcggccgctg actacaagga tgacgacgat aaatctagaa tggcttctaa ctttactcag
      121 ttcgttctcg tcgacaatgg cggaactggc gacgtgactg tcgccccaag caacttcgct
      181 aacgggatcg ctgaatggat cagctctaac tcgcgttcac aggcttacaa agtaacctgt
      241 agcgttcgtc agagctctgc gcagaatcgc aaatacacca tcaaagtcga ggtgcctaaa
      301 ggcgcctggc gttcgtactt aaatatggaa ctaaccattc caattttcgc cacgaattcc
      361 gactgcgagc ttattgttaa ggcaatgcaa ggtctcctaa aagatggaaa cccgattccc
      421 tcagcaatcg cagcaaactc cggcatctac gaggccagcg gttccggacg ggctgacgca
      481 ttggacgatt ttgatctgga tatgctggga agtgacgccc tcgatgattt tgaccttgac
      541 atgcttggtt cggatgccct tgatgacttt gacctcgaca tgctcggcag tgacgccctt
      601 gatgatttcg acctggacat gctgattaac tctagatgaa acccagcttt cttgtacaaa
      661 gtggtgagct ccgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
      721 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc
      781 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
      841 cccgtgccct gg
//
