LOCUS       sequence-89992-e         829 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Plasmid encoding multiplexed 4x gRNAs. This is a modified form of 
            the original plasmid described in the paper (Construct 19). mKate2 
            was replaced with dsRed2 because of distribution issues..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 829)
  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 829)
  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-gRNA3-28-gRNA4-28-gRNA5-28-gRNA6-28-pA
FEATURES             Location/Qualifiers
     source          1..829
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    205..224
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     misc_RNA        259..334
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    343..362
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     misc_RNA        395..470
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    479..498
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     misc_RNA        531..606
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    615..634
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     misc_RNA        667..742
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    751..770
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
ORIGIN
        1 agcgcaccga gggccgccac cacctgttcc tgtagtaaac cggtgattcg tcagtagggt
       61 tgtaaaggtt tttcttttcc tgagaaaaca accttttgtt ttctcaggtt ttgctttttg
      121 gcctttccct agctttaaaa aaaaaaaagc aaaactcacc gaggcagttc cataggatgg
      181 caagatcctg gtatcggtct gcgagttcac tgccgtatag gcagctaaga aagctagcgt
      241 gtactctctg aggtgctcgt tttagagcta gaaatagcaa gttaaaataa ggctagtccg
      301 ttatcaactt gaaaaagtgg caccgagtcg gtgctttttt tcgttcactg ccgtataggc
      361 agctaagaaa aggtgacgca gataagaacc agttgtttta gagctagaaa tagcaagtta
      421 aaataaggct agtccgttat caacttgaaa aagtggcacc gagtcggtgc tttttttcgt
      481 tcactgccgt ataggcagct aagaaacagg gcatcaagtc agccatcagc gttttagagc
      541 tagaaatagc aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt
      601 cggtgctttt tttcgttcac tgccgtatag gcagctaaga aaagtcggag tcaccctcct
      661 ggaaacgttt tagagctaga aatagcaagt taaaataagg ctagtccgtt atcaacttga
      721 aaaagtggca ccgagtcggt gctttttttc gttcactgcc gtataggcag ctaagaaacc
      781 cgggtaagcg ggactctggg gttcgaaatg accgaccaag cgacgccca
//
