LOCUS       sequence-130148-         810 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expression vector for sgRNA and Cas9 linked via T2A to BFP linked to
            the Ad4 E4orf6 gene via P2A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 810)
  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 810)
  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-BFP-P2A-Ad4E4orf6
FEATURES             Location/Qualifiers
     source          1..810
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(15..68)
                     /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(72..119)
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
ORIGIN
        1 ctcaccatgc tagcagggcc gggattctcc tccacgtcac cgcatgttag cagacttcct
       61 ctgccctcaa gctttttctt ttttgcctgg ccggcctttt tcgtggccgc cggccttttg
      121 tcgcctccca gctgagacag gtcgatccgt gtctcgtaca ggccggtgat gctctggtgg
      181 atcagggtgg cgtccagcac ctctttggtg ctggtgtacc tcttccggtc gatggtggtg
      241 tcaaagtact tgaaggcggc aggggctccc agattggtca gggtaaacag gtggatgata
      301 ttctcggcct gctctctgat gggcttatcc cggtgcttgt tgtaggcgga cagcactttg
      361 tccagattag cgtcggccag gatcactctc ttggagaact cgctgatctg ctcgatgatc
      421 tcgtccaggt agtgcttgtg ctgttccaca aacagctgtt tctgctcatt atcctcgggg
      481 gagcccttca gcttctcata gtggctggcc aggtacagga agttcacata tttggagggc
      541 agggccagtt cgtttccctt ctgcagttcg ccggcagagg ccagcattct cttccggccg
      601 ttttccagct cgaacaggga gtacttaggc agcttgatga tcaggtcctt tttcacttct
      661 ttgtagccct tggcttccag aaagtcgatg ggattcttct cgaagctgct tctttccatg
      721 atggtgatcc ccagcagctc tttcacactc ttcagtttct tggacttgcc cttttccact
      781 ttggccacca ccagcacaga ataggccacg
//
