LOCUS       sequence-69622-e         953 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses mammalian cell codon-optimized dCas9-VP64.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 953)
  AUTHORS   Maeder ML, Linder SJ, Cascio VM, Fu Y, Ho QH, Joung JK
  TITLE     CRISPR RNA-guided activation of endogenous human genes.
  JOURNAL   Nat Methods. 2013 Jul 25. doi: 10.1038/nmeth.2598.
  PUBMED    23892898
REFERENCE   2  (bases 1 to 953)
  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. 2013 Jul 25. doi: 10.1038/nmeth.2598."
COMMENT     Sequence Label: pMLM3705
FEATURES             Location/Qualifiers
     source          1..953
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(1..18)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(27..176)
                     /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(207..272)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             complement(279..299)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 atggtgatgg tgatgatgac cggttataac atatcgagat cgaaatcgtc cagagcatcg
       61 gagccgagca tgtccagatc aaagtcatcc aatgcgtcgc ttcccaacat atccaggtca
      121 aagtcatcga gggcatcaga acccagcatg tcgagatcga aatcgtccag cgcgtcggcg
      181 cgcccgcttc caccgcctcc tgcagccttg tcatcgtcat ccttgtaatc gatgtcatga
      241 tctttataat caccgtcatg gtctttgtag tcgctcgaga ctttcctctt cttcttgggg
      301 gatccgtcac ccccaagctg tgacaaatct atccgagttt catataatcc cgtgatggat
      361 tggtgaatca gtgtcgcgtc tagcacctcc ttggtagaag tgtatcgttt gcgatctatc
      421 gttgtgtcaa aatacttgaa tgcggctgga gcgccgaggt tggtaagagt aaacaaatgg
      481 ataatatttt ccgcctgctc acgtatgggt ttatccctgt gcttgttgta tgcgcttaat
      541 actttgtcca gattggcatc agctaggatg actctcttac tgaattccga aatttgctct
      601 atgatttcgt cgagataatg tttgtgctgc tcaacaaaaa gttgcttctg ttcgttatct
      661 tcaggtgaac ctttcaactt ctcgtaatgg gacgctaaat acaggaaatt cacgtattta
      721 gacggtagtg cgagttcgtt ccccttttga agctctccgg cgctagccaa catccgtttt
      781 cggccatttt ctaactcaaa cagactatac tttggtagtt taattatgag atcctttttt
      841 acttccttgt aacctttcgc ctcaaggaag tcgatggggt tcttttcaaa agacgagcgc
      901 tccataatcg ttatccccaa taattctttg actgacttca gtttcttgga ttt
//
