LOCUS       sequence-135432-         604 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expression of human codon-optimized dCas9-DNMT3A fusion with 
            T2A-PuroR for targeted DNA methylation in mammalian cells; cloning 
            backbone for sgRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 604)
  AUTHORS   Vojta A, Dobrinic P, Tadic V, Bockor L, Korac P, Julg B, Klasic M, 
            Zoldos V
  TITLE     Repurposing the CRISPR-Cas9 system for targeted DNA methylation.
  JOURNAL   Nucleic Acids Res. 2016 Mar 11. pii: gkw159.
  PUBMED    26969735
REFERENCE   2  (bases 1 to 604)
  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: "Nucleic 
            Acids Res. 2016 Mar 11. pii: gkw159."
COMMENT     Sequence Label: pdCas9-DNMT3A-PuroR
FEATURES             Location/Qualifiers
     source          1..604
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(310..330)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(337..402)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     regulatory      402..411
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 gctgtcgtcc accttggcca tctcgttgct gaagatctct tgcagatagc agatccggtt
       61 cttccgtctg gtgtatcttc ttctggcggt tctcttcagc cgggtggcct cggctgtttc
      121 gccgctgtcg aacagcaggg ctccgatcag gttcttcttg atgctgtgcc ggtcggtgtt
      181 gcccagcacc ttgaatttct tgctgggcac cttgtactcg tcggtgatca cggcccagcc
      241 cacagagttg gtgccgatgg ccaggccgat gctgtacttc ttgtcggctg ctgggactcc
      301 gtggataccg accttccgct tcttctttgg ggccatctta tcgtcatcgt ctttgtaatc
      361 aatatcatga tccttgtagt ctccgtcgtg gtccttatag tccatggtgg caccggtcca
      421 acctgaaaaa aagtgatttc aggcaggtgc tccaggtaat taaacattaa taccccacca
      481 accaaccatc ccttaaaccc ttacctcttg ctcagctaat tacagcccgg aggagaaggg
      541 ccgtcccgcc cgctcacctg tgggagtaac gcggtcagtc agagccgggg cgggcggcgc
      601 gagg
//
