LOCUS       sequence-69386-e         769 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses inactivated S. pyogenes dCas9 (D10A, H840A) in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 769)
  AUTHORS   Perez-Pinera P, Kocak DD, Vockley CM, Adler AF, Kabadi AM, Polstein 
            LR, Thakore PI, Glass KA, Ousterout DG, Leong KW, Guilak F, Crawford
            GE, Reddy TE, Gersbach CA
  TITLE     RNA-guided gene activation by CRISPR-Cas9-based transcription 
            factors.
  JOURNAL   Nat Methods. 2013 Jul 25. doi: 10.1038/nmeth.2600.
  PUBMED    23892895
REFERENCE   2  (bases 1 to 769)
  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.2600."
COMMENT     Sequence Label: pcDNA-dCas9
FEATURES             Location/Qualifiers
     source          1..769
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     48..67
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        48..66
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             92..157
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             164..184
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 actctctggc taactagaga acccactgct tactggctta tcgaaattaa tacgactcac
       61 tatagggaga cccaagctgg ctagcgccat ggactacaaa gaccatgacg gtgattataa
      121 agatcatgac atcgattaca aggatgacga tgacaagatg gcccccaaga agaagaggaa
      181 ggtgggccgc ggaatggata agaaatactc aataggctta gctatcggca caaatagcgt
      241 cggatgggcg gtgatcactg atgaatataa ggttccgtct aaaaagttca aggttctggg
      301 aaatacagac cgccacagta tcaaaaaaaa tcttataggg gctcttttat ttgacagtgg
      361 agagacagcg gaagcgactc gtctcaaacg gacagctcgt agaaggtata cacgtcggaa
      421 gaatcgtatt tgttatctac aggagatttt ttcaaatgag atggcgaaag tagatgatag
      481 tttctttcat cgacttgaag agtctttttt ggtggaagaa gacaagaagc atgaacgtca
      541 tcctattttt ggaaatatag tagatgaagt tgcttatcat gagaaatatc caactatcta
      601 tcatctgcga aaaaaattgg tagattctac tgataaagcg gatttgcgct taatctattt
      661 ggccttagcg catatgatta agtttcgtgg tcattttttg attgagggag atttaaatcc
      721 tgataatagt gatgtggaca aactatttat ccagttggta caaacctac
//
