LOCUS       sequence-154064-         848 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses dCas9-VP64 under actin promoter for SAM CRISPRa in 
            Drosophila cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 848)
  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 848)
  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:dCas9-VP64
FEATURES             Location/Qualifiers
     source          1..848
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      19..28
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    36..60
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(62..460)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     CDS             complement(461..514)
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             complement(536..685)
                     /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"
     CDS             complement(710..730)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 gctcctcgcc cttgctcacc atggtggcgg agctcaccac tttgtacaag aaagctgggt
       61 tttagccctc ccacacataa ccagagggca gcaattcacg aatcccaact gccgtcggct
      121 gtccatcact gtccttcact atggctttga tcccaggatg cagatcgaga agcacctgtc
      181 ggcaccgtcc gcaggggctc aagatgcccc tgttctcatt tccgatcgcg acgatacaag
      241 tcaggttgcc agctgccgca gcagcagcag tgcccagcac cacgagttct gcacaaggtc
      301 ccccagtaaa atgatataca ttgacaccag tgaagatgcg gccgtcgcta gagagagctg
      361 cgctggcgac gctgtagtct tcagagatgg ggatgctgtt gattgtagcc gttgctcttt
      421 caatgagggt ggattcttct tgagacaaag gcttggccat tgggccagga ttctcctcga
      481 cgtcaccgca tgttagcaga cttcctctgc cctctccact gcctgtacag ttaatcagca
      541 tgtccaggtc gaaatcatca agggcgtcac tgccgagcat gtcgaggtca aagtcatcaa
      601 gggcatccga accaagcatg tcaaggtcaa aatcatcgag ggcgtcactt cccagcatat
      661 ccagatcaaa atcgtccaat gcgtcagccc gtccggatcc agcggccgcc accttcctct
      721 ttttcttagg tccgctacct cctcctccgc ttcctcctcc tccgcttcca cctcctccag
      781 cgctgtcgcc tcccagctga gacaggtcga tccgtgtctc gtacaggccg gtgatgctct
      841 ggtggatc
//
