LOCUS       sequence-114398-         649 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expression vector for sgRNAs cloned into the BbsI sites and for 
            expression of Cas9 linked to BFP via a T2A peptide.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 649)
  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 649)
  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
FEATURES             Location/Qualifiers
     source          1..649
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             272..319
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             323..376
                     /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(428..449)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
ORIGIN
        1 gagagtgatc ctggccgacg ctaatctgga caaagtgctg tccgcctaca acaagcaccg
       61 ggataagccc atcagagagc aggccgagaa tatcatccac ctgtttaccc tgaccaatct
      121 gggagcccct gccgccttca agtactttga caccaccatc gaccggaaga ggtacaccag
      181 caccaaagag gtgctggacg ccaccctgat ccaccagagc atcaccggcc tgtacgagac
      241 acggatcgac ctgtctcagc tgggaggcga caaaaggccg gcggccacga aaaaggccgg
      301 ccaggcaaaa aagaaaaagc ttgagggcag aggaagtctg ctaacatgcg gtgacgtgga
      361 ggagaatccc ggccctgcta gcatggtgag caagggcgag gagctgttca ccggggtggt
      421 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgaggggcga
      481 gggcgagggc gatgccacca acggcaagct gaccctgaag ttcatctgca ccaccggcaa
      541 gctgcccgtg ccctggccca ccctcgtgac caccctgagc cacggcgtgc agtgcttcgc
      601 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatg
//
