LOCUS       sequence-114397-         436 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expression vector for sgRNAs cloned into the BbsI sites, shRNAs into
            BamHI.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 436)
  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 436)
  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-H1-(BamHI)
FEATURES             Location/Qualifiers
     source          1..436
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(252..272)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(279..344)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     regulatory      344..353
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 ttcttccgtc tggtgtatct tcttctggcg gttctcttca gccgggtggc ctcggctgtt
       61 tcgccgctgt cgaacagcag ggctccgatc aggttcttct tgatgctgtg ccggtcggtg
      121 ttgcccagca ccttgaattt cttgctgggc accttgtact cgtcggtgat cacggcccag
      181 cccacagagt tggtgccgat gtccaggccg atgctgtact tcttgtcggc tgctgggact
      241 ccgtggatac cgaccttccg cttcttcttt ggggccatct tatcgtcatc gtctttgtaa
      301 tcaatatcat gatccttgta gtctccgtcg tggtccttat agtccatggt ggcaccggtc
      361 caacctgaaa aaaagtgatt tcaggcaggt gctccaggta attaaacatt aataccccac
      421 caaccaacca tccctt
//
