LOCUS       sequence-106318-         833 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Mammalian CRISPR-SP-Cas9 reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 833)
  AUTHORS   D'Astolfo DS, Pagliero RJ, Pras A, Karthaus WR, Clevers H, Prasad V,
            Lebbink RJ, Rehmann H, Geijsen N
  TITLE     Efficient Intracellular Delivery of Native Proteins.
  JOURNAL   Cell. 2015 Apr 23;161(3):674-690. doi: 10.1016/j.cell.2015.03.028.
  PUBMED    25910214
REFERENCE   2  (bases 1 to 833)
  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"; doi: 
            "10.1016/j.cell.2015.03"; journalName: "Cell"; date: "2015-04-23- 
            23"; volume: "161"; issue: "3"; pages: "674-690"
COMMENT     Sequence Label: CRISPR-SP-Cas9 reporter
FEATURES             Location/Qualifiers
     source          1..833
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      20..29
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             35..61
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             117..815
                     /codon_start=1
                     /product="dimeric variant of DsRed fluorescent protein 
                     (Shaner et al., 2004)"
                     /label=dTomato
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGL
                     VTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIHQ
                     ALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRHH
                     LFLYGMDELYK"
     primer_bind     complement(247..265)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     515..534
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     709..732
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
ORIGIN
        1 tttcaggtgt cgtgaagcgg ccgccatggg atcttaccct tatgatgtgc ctgactacgc
       61 cggaggagga ggatctgggg ccactaggga caggatcgga gctccggccg cataaggtga
      121 gcaagggcga ggaggtcatc aaagagttca tgcgcttcaa ggtgcgcatg gagggctcca
      181 tgaacggcca cgagttcgag atcgagggcg agggcgaggg ccgcccctac gagggcaccc
      241 agaccgccaa gctgaaggtg accaagggcg gccccctgcc cttcgcctgg gacatcctgt
      301 ccccccagtt catgtacggc tccaaggcgt acgtgaagca ccccgccgac atccccgatt
      361 acaagaagct gtccttcccc gagggcttca agtgggagcg cgtgatgaac ttcgaggacg
      421 gcggtctggt gaccgtgacc caggactcct ccctgcagga cggcacgctg atctacaagg
      481 tgaagatgcg cggcaccaac ttcccccccg acggccccgt aatgcagaag aagaccatgg
      541 gctgggaggc ctccaccgag cgcctgtacc cccgcgacgg cgtgctgaag ggcgagatcc
      601 accaggccct gaagctgaag gacggcggcc actacctggt ggagttcaag accatctaca
      661 tggccaagaa gcccgtgcaa ctgcccggct actactacgt ggacaccaag ctggacatca
      721 cctcccacaa cgaggactac accatcgtgg aacagtacga gcgctccgag ggccgccacc
      781 acctgttcct gtacggcatg gacgagctgt acaagtaagg atccctcccc ccc
//
