LOCUS       sequence-110999-         765 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 765)
  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 765)
  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..765
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(join(183..695,696..698,699..701))
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLSHGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGVDFKEDGNILGHKLEYNFNSHNIYIMAVKQKNGIK
                     VNFKIRHNVE"
     primer_bind     635..656
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(708..761)
                     /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"
ORIGIN
        1 atgccgagag tgatcccggc ggcggtgcgg aactccagca ggaccatgtg atcgcgcttc
       61 tcgttggggt ctttgctcag cacggactgg gtgctcaggt agtggctgtc gggcagcagc
      121 acggggccgt cgccgatggg ggtgttctgc tggtagtggt cggcgagctg cacgctgccg
      181 tcctccacgt tgtggcggat cttgaagttc accttgatgc cgttcttctg cttgacggcc
      241 atgatataga tgttgtggct gttgaagttg tactccagct tgtgccccag gatgttgccg
      301 tcctccttga agtcgacgcc cttcagctcg atgcggttca ccagggtgtc gccctcgaac
      361 ttcacctcgg cgcgggtctt gtaggtgccg tcgtccttga agaagatggt gcgctcctgg
      421 acgtagcctt cgggcatggc ggacttgaag aagtcgtgct gcttcatgtg gtcggggtag
      481 cgggcgaagc actgcacgcc gtggctcagg gtggtcacga gggtgggcca gggcacgggc
      541 agcttgccgg tggtgcagat gaacttcagg gtcagcttgc cgttggtggc atcgccctcg
      601 ccctcgcccc tcacgctgaa cttgtggccg tttacgtcgc cgtccagctc gaccaggatg
      661 ggcaccaccc cggtgaacag ctcctcgccc ttgctcacca tgctagcagg gccgggattc
      721 tcctccacgt caccgcatgt tagcagactt cctctgccct caagc
//
