LOCUS       sequence-122242-         824 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  expresses VP64-dCas9-VP64, same plasmid as Addgene 59791 but with 
            LoxP sites removed.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 824)
  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  JOURNAL   Unpublished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 824)
  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            "Unpublished"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 hUbC- VP64-dCas9-VP64-T2A- GFP(No LoxP sites)
FEATURES             Location/Qualifiers
     source          1..824
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(14..67)
                     /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(77..226)
                     /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(239..259)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 tgctcacggt accagggccg ggattctcct ccacgtcacc gcatgttaga agacttcctc
       61 tgccctcgct agcgattaac atatcgagat cgaaatcgtc cagagcatcg gagccgagca
      121 tgtccagatc aaagtcatcc aatgcgtcgc ttcccaacat atccaggtca aagtcatcga
      181 gggcatcaga acccagcatg tcgagatcga aatcgtccag cgcgtcggcg cggctagcca
      241 ccttcctctt cttcttgggg tcagccctgc tgtctccacc gagctgagag aggtcgattc
      301 ttgtttcata gagccccgta attgactgat gaatcagtgt ggcgtccagg acctcctttg
      361 tagaggtgta ccgctttctg tctatggtgg tgtcgaagta cttgaaggct gcaggcgcgc
      421 ccaagttggt cagagtaaac aagtggataa tgttttctgc ctgctccctg atgggcttat
      481 ccctgtgctt attgtaagca gaaagcacct tatcgaggtt agcgtcggcg aggatcactc
      541 ttttggagaa ttcgcttatt tgctcgatga tctcatcaag gtagtgtttg tgttgttcca
      601 cgaacagctg cttctgctca ttatcttcgg gagacccttt gagcttttca tagtggctgg
      661 ccagatacaa gaaattaacg tatttagagg gcagtgccag ctcgttacct ttctgcagct
      721 cgcccgcact agcgagcatt cgtttccggc cgttttcaag ctcaaagaga gagtacttgg
      781 gaagcttaat gatgaggtct tttttgacct ctttatatcc tttc
//
