LOCUS       sequence-154039-         755 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expression vector for sgRNA and for Expression of Cas9 linked to BFP
            via T2A linked to Ad4 E1B via P2A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 755)
  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 755)
  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-Ad4E1B
FEATURES             Location/Qualifiers
     source          1..755
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     126..145
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        222..297
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        394..679
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 aggctgttag agagataatt ggaattaatt tgactgtaaa cacaaagata ttagtacaaa
       61 atacgtgacg tagaaagtaa taatttcttg ggtagtttgc agttttaaaa ttatgtttta
      121 aaatggacta tcatatgctt accgtaactt gaaagtattt cgatttcttg gctttatata
      181 tcttgtggaa aggacgaaac accgggtctt cgagaagacc tgttttagag ctagaaatag
      241 caagttaaaa taaggctagt ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt
      301 tttgttttag agctagaaat agcaagttaa aataaggcta gtccgttttt agcgcgtgcg
      361 ccaattctgc agacaaatgg ctctagaggt acccgttaca taacttacgg taaatggccc
      421 gcctggctga ccgcccaacg acccccgccc attgacgtca atagtaacgc caatagggac
      481 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
      541 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
      601 gcattgtgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
      661 agtcatcgct attaccatgg tcgaggtgag ccccacgttc tgcttcactc tccccatctc
      721 ccccccctcc ccacccccaa ttttgtattt attta
//
