LOCUS       sequence-112717-         724 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expression vector for sgRNA and for Expression of Cas9 linked to BFP
            via T2A linked to Ad4 E1B via P2A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 724)
  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 724)
  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-Ad4E1B
FEATURES             Location/Qualifiers
     source          1..724
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             267..314
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             318..371
                     /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(423..444)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
ORIGIN
        1 tgatcctggc cgacgctaat ctggacaaag tgctgtccgc ctacaacaag caccgggata
       61 agcccatcag agagcaggcc gagaatatca tccacctgtt taccctgacc aatctgggag
      121 cccctgccgc cttcaagtac tttgacacca ccatcgaccg gaagaggtac accagcacca
      181 aagaggtgct ggacgccacc ctgatccacc agagcatcac cggcctgtac gagacacgga
      241 tcgacctgtc tcagctggga ggcgacaaaa ggccggcggc cacgaaaaag gccggccagg
      301 caaaaaagaa aaagcttgag ggcagaggaa gtctgctaac atgcggtgac gtggaggaga
      361 atcccggccc tgctagcatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
      421 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgagg ggcgagggcg
      481 agggcgatgc caccaacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc
      541 ccgtgccctg gcccaccctc gtgaccaccc tgagccacgg cgtgcagtgc ttcgcccgct
      601 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
      661 aggagcgcac catcttcttc aaggacgacg gcacctacaa gacccgcgcc gaggtgaagt
      721 tcga
//
