LOCUS       sequence-89911-e         788 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Plasmid encoding the Triplex/gRNA architecture.This is a modified 
            form of the original plasmid described in the paper (Construct 3). 
            mKate2 was replaced with dsRed2 because of distribution issues..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 788)
  AUTHORS   Nissim L, Perli SD, Fridkin A, Perez-Pinera P, Lu TK
  TITLE     Multiplexed and Programmable Regulation of Gene Networks with an 
            Integrated RNA and CRISPR/Cas Toolkit in Human Cells.
  JOURNAL   Mol Cell. 2014 May 14. pii: S1097-2765(14)00355-4. doi: 
            10.1016/j.molcel.2014.04.022.
  PUBMED    24837679
REFERENCE   2  (bases 1 to 788)
  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. 
            2014 May 14. pii: S1097-2765(14)00355-4. doi: 
            10.1016/j.molcel.2014.04.022."
COMMENT     Sequence Label: CMVp-dsRed2-Triplex-28-gRNA1-28-pA
FEATURES             Location/Qualifiers
     source          1..788
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     polyA_signal    165..212
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     237..256
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     protein_bind    complement(519..538)
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     misc_RNA        complement(547..622)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(651..670)
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
ORIGIN
        1 tatggcaggg cctgccgccc cgacgttggc tgcgagccct gggccttcac ccgaacttgg
       61 ggggtggggt ggggaaaagg aagaaacgcg ggcgtattgg ccccaatggc gtctcggtgg
      121 ggtatcgaca gagtgccagc cctgggaccg aaccccgcgt ttatgaacaa acgacccaac
      181 accgtgcgtt ttattctgtc tttttattgc cgtcatagcg cgggttcctt ccggtattgt
      241 ctccttccgt gtttcagtta gcctccccct agggtgggcg aagaactcca gcatgagatc
      301 cccgcgctgg aggatcatcc agccggcgtc ccggaaaacg attccgaagc ccaacctttc
      361 atagaaggcg gcggtggaat cgaaatctcg tgatggcagg ttgggcgtcg cttggtcggt
      421 catttcgaac cccagagtcc cgcttagcta gcatcccggg gttgatgcgc tggcagtgtt
      481 cctgcgccgg ttgcattcga ttcctgtttg tttcttagct gcctatacgg cagtgaacga
      541 aaaaaagcac cgactcggtg ccactttttc aagttgataa cggactagcc ttattttaac
      601 ttgctatttc tagctctaaa actgcttcgc tacacgcgac tatttcttag ctgcctatac
      661 ggcagtgaac tcgcagacca ataccaggat cttgccatcc tatggaactg cctcggtgag
      721 ttttgctttt ttttttttaa agctagggaa aggccaaaaa gcaaaacctg agaaaacaaa
      781 aggttgtt
//
