LOCUS       sequence-116722-         750 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 750)
  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  JOURNAL   Unpublished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 750)
  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            "Unpublished"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..750
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(26..79)
                     /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(89..238)
                     /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(251..271)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 gctcctcgcc cttgctcacg gtaccagggc cgggattctc ctccacgtca ccgcatgtta
       61 gaagacttcc tctgccctcg ctagcgatta acatatcgag atcgaaatcg tccagagcat
      121 cggagccgag catgtccaga tcaaagtcat ccaatgcgtc gcttcccaac atatccaggt
      181 caaagtcatc gagggcatca gaacccagca tgtcgagatc gaaatcgtcc agcgcgtcgg
      241 cgcggctagc caccttcctc ttcttcttgg ggtcagccct gctgtctcca ccgagctgag
      301 agaggtcgat tcttgtttca tagagccccg taattgactg atgaatcagt gtggcgtcca
      361 ggacctcctt tgtagaggtg taccgctttc tgtctatggt ggtgtcgaag tacttgaagg
      421 ctgcaggcgc gcccaagttg gtcagagtaa acaagtggat aatgttttct gcctgctccc
      481 tgatgggctt atccctgtgc ttattgtaag cagaaagcac cttatcgagg ttagcgtcgg
      541 cgaggatcac tcttttggag aattcgctta tttgctcgat gatctcatca aggtagtgtt
      601 tgtgttgttc cacgaacagc tgcttctgct cattatcttc gggagaccct ttgagctttt
      661 catagtggct ggccagatac aagaaattaa cgtatttaga gggcagtgcc agctcgttac
      721 ctttctgcag ctcgcccgca ctagcgagca
//
