LOCUS       sequence-190279-        1244 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Empty plasmid backbone to express 2 sgRNAs (use BbsI and BsmBI for 
            cloning).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1244)
  AUTHORS   Billon P, Bryant EE, Joseph SA, Nambiar TS, Hayward SB, Rothstein R,
            Ciccia A
  TITLE     CRISPR-Mediated Base Editing Enables Efficient Disruption of 
            Eukaryotic Genes through Induction of STOP Codons.
  JOURNAL   Mol Cell. 2017 Sep 4. pii: S1097-2765(17)30605-6. doi: 
            10.1016/j.molcel.2017.08.008.
  PUBMED    28890334
REFERENCE   2  (bases 1 to 1244)
  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: "Mol Cell. 
            2017 Sep 4. pii: S1097-2765(17)30605-6. doi: 
            10.1016/j.molcel.2017.08.008."
COMMENT     Sequence Label: B52 (empty plasmid backbone to express 2 sgRNAs)
FEATURES             Location/Qualifiers
     source          1..1244
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        complement(94..342)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        complement(356..431)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(451..699)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     promoter        806..910
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tgtgtgacta gcgtgcactt tttcagttga taacggacta gccttatttt aacttgctat
       61 ttctagctct aaaacaggtc ttctcgaaga cccggtgttt cgtcctttcc acaagatata
      121 taaagccaag aaatcgaaat actttcaagt tacggtaagc atatgatagt ccattttaaa
      181 acataatttt aaaactgcaa actacccaag aaattattac tttctacgtc acgtattttg
      241 tactaatatc tttgtgttta cagtcaaatt aattccaatt atctctctaa cagccttgta
      301 tcgtatatgc aaatatgaag gaatcatggg aaataggccc tcaagcttaa aaaaagcacc
      361 gactcggtgc cactttttca agttgataac ggactagcct tattttaact tgctatttct
      421 agctctaaaa cgagacgcat taatcgtctc ggtgtttcgt cctttccaca agatatataa
      481 agccaagaaa tcgaaatact ttcaagttac ggtaagcata tgatagtcca ttttaaaaca
      541 taattttaaa actgcaaact acccaagaaa ttattacttt ctacgtcacg tattttgtac
      601 taatatcttt gtgtttacag tcaaattaat tctaattatc tctctaacag ccttgtatcg
      661 tatatgcaaa tatgaaggaa tcatgggaaa taggccctct tcctgcccga ccttggtacc
      721 ggatccagtc gactgaattg gttcctttaa agcctgcttt tttgtacagc tagcgacgtc
      781 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
      841 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
      901 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
      961 ttgccttcct gttttgctca ccagaaacgc tggtgaaagt aaagatgctg agatcagttg
     1021 ggtgcacgag tgggttacat cgaactggat ctcacagcgg tagatccttg agagtttcgc
     1081 ccgagacgtt tccatgatga gcactttaaa gtctgctatg tggcgcgtat tatccgtatg
     1141 acgcgggcag agcactcgtc gccgcataac ctattctcag aatgacttgt tgagtactca
     1201 cagtccagga aagcatctta cgattgcatg aacgtaagag aata
//
