LOCUS       sequence-112724-         683 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expression vector for sgRNA and for Expression of Cas9 linked via 
            T2A to mCherry and to Ad4 E1B55K via P2A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 683)
  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 683)
  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-Ad4E1B
FEATURES             Location/Qualifiers
     source          1..683
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     98..117
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        194..269
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        366..651
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 tttgactgta aacacaaaga tattagtaca aaatacgtga cgtagaaagt aataatttct
       61 tgggtagttt gcagttttaa aattatgttt taaaatggac tatcatatgc ttaccgtaac
      121 ttgaaagtat ttcgatttct tggctttata tatcttgtgg aaaggacgaa acaccgggtc
      181 ttcgagaaga cctgttttag agctagaaat agcaagttaa aataaggcta gtccgttatc
      241 aacttgaaaa agtggcaccg agtcggtgct tttttgtttt agagctagaa atagcaagtt
      301 aaaataaggc tagtccgttt ttagcgcgtg cgccaattct gcagacaaat ggctctagag
      361 gtacccgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      421 ccattgacgt caatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
      481 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct
      541 attgacgtca atgacggtaa atggcccgcc tggcattgtg cccagtacat gaccttatgg
      601 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtcgaggtg
      661 agccccacgt tctgcttcac tct
//
