LOCUS       sequence-96696-e         576 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Co-expresses human optimized S. pyogenes dCas9 fused to two copies 
            of VP64 and GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 576)
  AUTHORS   Kabadi AM, Ousterout DG, Hilton IB, Gersbach CA
  TITLE     Multiplex CRISPR/Cas9-based genome engineering from a single 
            lentiviral vector.
  JOURNAL   Nucleic Acids Res. 2014 Aug 13. pii: gku749.
  PUBMED    25122746
REFERENCE   2  (bases 1 to 576)
  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. 2014 Aug 13. pii: gku749."
COMMENT     Sequence Label: pLV hUbC-VP64 dCas9 VP64-T2A-GFP
FEATURES             Location/Qualifiers
     source          1..576
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(13..66)
                     /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(76..225)
                     /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(238..258)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 gctcacggta ccagggccgg gattctcctc cacgtcaccg catgttagaa gacttcctct
       61 gccctcgcta gcgattaaca tatcgagatc gaaatcgtcc agagcatcgg agccgagcat
      121 gtccagatca aagtcatcca atgcgtcgct tcccaacata tccaggtcaa agtcatcgag
      181 ggcatcagaa cccagcatgt cgagatcgaa atcgtccagc gcgtcggcgc ggctagccac
      241 cttcctcttc ttcttggggt cagccctgct gtctccaccg agctgagaga ggtcgattct
      301 tgtttcatag agccccgtaa ttgactgatg aatcagtgtg gcgtccagga cctcctttgt
      361 agaggtgtac cgctttctgt ctatggtggt gtcgaagtac ttgaaggctg caggcgcgcc
      421 caagttggtc agagtaaaca agtggataat gttttctgcc tgctccctga tgggcttatc
      481 cctgtgctta ttgtaagcag aaagcacctt atcgaggtta gcgtcggcga ggatcactct
      541 tttggagaat tcgcttattt gctcgatgat ctcatc
//
