LOCUS       sequence-110992-         706 bp    DNA     linear   UNA 14-APR-2025
DEFINITION  Expression vector for sgRNA and for Expression of Cas9 linked via 
            T2A to mCherry and to Ad4 E1B55K via P2A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 706)
  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 706)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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-Ad4E1B
FEATURES             Location/Qualifiers
     source          1..706
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             38..85
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             89..142
                     /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"
     primer_bind     complement(294..312)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
ORIGIN
        1 cgagacacgg atcgacctgt ctcagctggg aggcgacaaa aggccggcgg ccacgaaaaa
       61 ggccggccag gcaaaaaaga aaaagcttga gggcagagga agtctgctaa catgcggtga
      121 cgtggaggag aatcccggcc ctgctagcat ggtgagcaag ggcgaggagg ataacatggc
      181 catcatcaag gagttcatgc gcttcaaggt gcacatggag ggctccgtga acggccacga
      241 gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct
      301 gaaggtgacc aagggtggcc ccctgccctt cgcctgggac atcctgtccc ctcagttcat
      361 gtacggctcc aaggcctacg tgaagcaccc cgccgacatc cccgactact tgaagctgtc
      421 cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac
      481 cgtgacccag gactcctccc tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg
      541 caccaacttc ccctcagacg gccccgtaat gcagaagaaa accatgggct gggaggcctc
      601 ctccgagcgg atgtaccccg aggacggcgc cctgaagggc gagatcaagc agaggctgaa
      661 gctgaaggac ggcggccact acgacgctga ggtcaagacc acctac
//
