LOCUS       sequence-116720-         659 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  expresses a single gRNA and VP64-dCas9-VP64.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 659)
  AUTHORS   Gersbach C
  TITLE     Multiplex CRISPR/Cas9 Genome Engineering for Directing Myogenic 
            Cellular Differentiation in Primary Human Cells
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 659)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pLV hU6-gRNA(anti-sense)
            hUbC-VP64-dCas9-VP64-...
FEATURES             Location/Qualifiers
     source          1..659
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      133..142
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             142..207
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             223..369
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation 
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label=VP64
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDM"
     CDS             376..396
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 aggcaccttt tgaaatgtaa tcatttgggt caatatgtaa ttttcagtgt tagactagta
       61 aattgtccgc taaattctgg ccgtttttgg cttttttgtt agacgaagct tgggctgcag
      121 gtcgactcta gagccaccat ggactacaaa gaccatgacg gtgattataa agatcatgac
      181 atcgattaca aggatgacga tgacaagcac ggtccgcggg ctgacgcatt ggacgatttt
      241 gatctggata tgctgggaag tgacgccctc gatgattttg accttgacat gcttggttcg
      301 gatgcccttg atgactttga cctcgacatg ctcggcagtg acgcccttga tgatttcgac
      361 ctggacatgg ttaaccccaa gaagaagagg aaggtgggcc gcggaatgga caagaagtac
      421 tccattgggc tcgccatcgg cacaaacagc gtcggctggg ccgtcattac ggacgagtac
      481 aaggtgccga gcaaaaaatt caaagttctg ggcaataccg atcgccacag cataaagaag
      541 aacctcattg gcgccctcct gttcgactcc ggggaaaccg ccgaagccac gcggctcaaa
      601 agaacagcac ggcgcagata tacccgcaga aagaatcgga tctgctacct gcaggagat
//
