LOCUS       sequence-114401-         667 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expression vector for sgRNA and for Expression of Cas9 linked via 
            T2A to mCherry linked to the Ad4 E4orf6 gene via P2A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 667)
  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 667)
  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-mcherry-P2A-Ad4E4orf6
FEATURES             Location/Qualifiers
     source          1..667
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             282..329
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             333..386
                     /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(538..556)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
ORIGIN
        1 agttctccaa gagagtgatc ctggccgacg ctaatctgga caaagtgctg tccgcctaca
       61 acaagcaccg ggataagccc atcagagagc aggccgagaa tatcatccac ctgtttaccc
      121 tgaccaatct gggagcccct gccgccttca agtactttga caccaccatc gaccggaaga
      181 ggtacaccag caccaaagag gtgctggacg ccaccctgat ccaccagagc atcaccggcc
      241 tgtacgagac acggatcgac ctgtctcagc tgggaggcga caaaaggccg gcggccacga
      301 aaaaggccgg ccaggcaaaa aagaaaaagc ttgagggcag aggaagtctg ctaacatgcg
      361 gtgacgtgga ggagaatccc ggccctgcta gcatggtgag caagggcgag gaggataaca
      421 tggccatcat caaggagttc atgcgcttca aggtgcacat ggagggctcc gtgaacggcc
      481 acgagttcga gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca
      541 agctgaaggt gaccaagggt ggccccctgc ccttcgcctg ggacatcctg tcccctcagt
      601 tcatgtacgg ctccaaggcc tacgtgaagc accccgccga catccccgac tacttgaagc
      661 tgtcctt
//
