LOCUS       sequence-89914-e         786 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 786)
  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 786)
  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..786
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      22..31
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             28..705
                     /codon_start=1
                     /product="improved tetrameric variant of DsRed fluorescent 
                     protein"
                     /label=DsRed2
                     /note="mammalian codon-optimized"
                     /translation="MASSENVITEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTVK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     ATVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGETHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDAKLDITSHNEDYTIVEQYERTEGRHH
                     LFL"
     primer_bind     complement(208..228)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     620..643
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
ORIGIN
        1 tgaaccgtca gatccgagct cgccaccatg gcctcctccg agaacgtcat caccgagttc
       61 atgcgcttca aggtgcgcat ggagggcacc gtgaacggcc acgagttcga gatcgagggc
      121 gagggcgagg gccgccccta cgagggccac aacaccgtga agctgaaggt gaccaagggc
      181 ggccccctgc ccttcgcctg ggacatcctg tccccccagt tccagtacgg ctccaaggtg
      241 tacgtgaagc accccgccga catccccgac tacaagaagc tgtccttccc cgagggcttc
      301 aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg cgaccgtgac ccaggactcc
      361 tccctgcagg acggctgctt catctacaag gtgaagttca tcggcgtgaa cttcccctcc
      421 gacggccccg tgatgcagaa gaagaccatg ggctgggagg cctccaccga gcgcctgtac
      481 ccccgcgacg gcgtgctgaa gggcgagacc cacaaggccc tgaagctgaa ggacggcggc
      541 cactacctgg tggagttcaa gtccatctac atggccaaga agcccgtgca gctgcccggc
      601 tactactacg tggacgccaa gctggacatc acctcccaca acgaggacta caccatcgtg
      661 gagcagtacg agcgcaccga gggccgccac cacctgttcc tgtagtaaac cggtgattcg
      721 tcagtagggt tgtaaaggtt tttcttttcc tgagaaaaca accttttgtt ttctcaggtt
      781 ttgctt
//
