LOCUS       sequence-112721-         650 bp    DNA     linear   UNA 14-APR-2025
DEFINITION  Expression vector for sgRNAs cloned into the BbsI sites and for 
            expression of Cas9 linked to BFP via a T2A peptide.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 650)
  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 650)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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
FEATURES             Location/Qualifiers
     source          1..650
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     86..105
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        182..257
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        354..639
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 cacaaagata ttagtacaaa atacgtgacg tagaaagtaa taatttcttg ggtagtttgc
       61 agttttaaaa ttatgtttta aaatggacta tcatatgctt accgtaactt gaaagtattt
      121 cgatttcttg gctttatata tcttgtggaa aggacgaaac accgggtctt cgagaagacc
      181 tgttttagag ctagaaatag caagttaaaa taaggctagt ccgttatcaa cttgaaaaag
      241 tggcaccgag tcggtgcttt tttgttttag agctagaaat agcaagttaa aataaggcta
      301 gtccgttttt agcgcgtgcg ccaattctgc agacaaatgg ctctagaggt acccgttaca
      361 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
      421 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
      481 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat
      541 gacggtaaat ggcccgcctg gcattgtgcc cagtacatga ccttatggga ctttcctact
      601 tggcagtaca tctacgtatt agtcatcgct attaccatgg tcgaggtgag
//
