LOCUS       sequence-130200-         754 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expression vector for sgRNA and Cas9 linked via T2A to BFP linked to
            the Ad4 E4orf6 gene via P2A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 754)
  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 754)
  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-Ad4E4orf6
FEATURES             Location/Qualifiers
     source          1..754
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     120..139
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        216..291
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        388..673
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 ttagagagat aattggaatt aatttgactg taaacacaaa gatattagta caaaatacgt
       61 gacgtagaaa gtaataattt cttgggtagt ttgcagtttt aaaattatgt tttaaaatgg
      121 actatcatat gcttaccgta acttgaaagt atttcgattt cttggcttta tatatcttgt
      181 ggaaaggacg aaacaccggg tcttcgagaa gacctgtttt agagctagaa atagcaagtt
      241 aaaataaggc tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg cttttttgtt
      301 ttagagctag aaatagcaag ttaaaataag gctagtccgt ttttagcgcg tgcgccaatt
      361 ctgcagacaa atggctctag aggtacccgt tacataactt acggtaaatg gcccgcctgg
      421 ctgaccgccc aacgaccccc gcccattgac gtcaatagta acgccaatag ggactttcca
      481 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
      541 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcattg
      601 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
      661 cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca tctccccccc
      721 ctccccaccc ccaattttgt atttatttat tttt
//
