LOCUS       sequence-155747-         750 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expression vector for sgRNA and for Expression of Cas9 linked via 
            T2A to mCherry linked to the Ad4 E4orf6 gene via P2A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 750)
  AUTHORS   Chu VT, Weber T, Wefers B, Wurst W, Sander S, Rajewsky K, Kuhn R
  TITLE     Increasing the efficiency of homology-directed repair for 
            CRISPR-Cas9-induced precise gene editing in mammalian cells.
  JOURNAL   Nat Biotechnol. 2015 Mar 24. doi: 10.1038/nbt.3198.
  PUBMED    25803306
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: "Nat 
            Biotechnol. 2015 Mar 24. doi: 10.1038/nbt.3198."
COMMENT     Sequence Label: pU6-(BbsI)_CBh-Cas9-T2A-mcherry-P2A-Ad4E4orf6
FEATURES             Location/Qualifiers
     source          1..750
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      28..37
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             37..102
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             109..129
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 aatcactttt tttcaggttg gaccggtgcc accatggact ataaggacca cgacggagac
       61 tacaaggatc atgatattga ttacaaagac gatgacgata agatggcccc aaagaagaag
      121 cggaaggtcg gtatccacgg agtcccagca gccgacaaga agtacagcat cggcctggac
      181 atcggcacca actctgtggg ctgggccgtg atcaccgacg agtacaaggt gcccagcaag
      241 aaattcaagg tgctgggcaa caccgaccgg cacagcatca agaagaacct gatcggagcc
      301 ctgctgttcg acagcggcga aacagccgag gccacccggc tgaagagaac cgccagaaga
      361 agatacacca gacggaagaa ccggatctgc tatctgcaag agatcttcag caacgagatg
      421 gccaaggtgg acgacagctt cttccacaga ctggaagagt ccttcctggt ggaagaggat
      481 aagaagcacg agcggcaccc catcttcggc aacatcgtgg acgaggtggc ctaccacgag
      541 aagtacccca ccatctacca cctgagaaag aaactggtgg acagcaccga caaggccgac
      601 ctgcggctga tctatctggc cctggcccac atgatcaagt tccggggcca cttcctgatc
      661 gagggcgacc tgaaccccga caacagcgac gtggacaagc tgttcatcca gctggtgcag
      721 acctacaacc agctgttcga ggaaaacccc
//
