LOCUS       sequence-138282-        1060 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  C-terminal tagging with mAID-mCherry2 using CRISPR/Cas9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1060)
  AUTHORS   Natsume T, Kiyomitsu T, Saga Y, Kanemaki MT
  TITLE     Rapid Protein Depletion in Human Cells by Auxin-Inducible Degron 
            Tagging with Short Homology Donors.
  JOURNAL   Cell Rep. 2016 Apr 5;15(1):210-8. doi: 10.1016/j.celrep.2016.03.001.
            Epub 2016 Mar 24.
  PUBMED    27052166
REFERENCE   2  (bases 1 to 1060)
  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.celrep.2016.03.001"; journalName: "Cell Rep"; date: 
            "2016-04-5- 5"; volume: "15"; issue: "1"; pages: "210-8"
COMMENT     Sequence Label: pMK294 (mAID-mCherry2-Bsr)
FEATURES             Location/Qualifiers
     source          1..1060
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     7..26
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7..25
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             62..265
                     /codon_start=1
                     /product="minimal degron from the auxin-responsive IAA17 
                     protein of Arabidopsis thaliana (Kubota et al., 2013)"
                     /label=mini-AID
                     /note="acts as an auxin-inducible degron (AID) in the 
                     presence of TIR1"
                     /translation="KEKSACPKDPAKPPAKAQVVGWPPVRSYRKNVMVSCQKSSGGPEA
                     AAFVKVSMDGAPYLRKIDLRMYK"
     CDS             272..982
                     /codon_start=1
                     /product="derivative of the monomeric red fluorescent 
                     protein mCherry"
                     /label=mCherry2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFNWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQCRTMGWEASTERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVDIKLDILSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(417..435)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
ORIGIN
        1 cgcgcgtaat acgactcact atagggcgaa ttggagctcc ccgggatccg gtgcaggcgc
       61 caaggagaag agtgcttgtc ctaaagatcc agccaaacct ccggccaagg cacaagttgt
      121 gggatggcca ccggtgagat cataccggaa gaacgtgatg gtttcctgcc aaaaatcaag
      181 cggtggcccg gaggcggcgg cgttcgtgaa ggtatcaatg gacggagcac cgtacttgag
      241 gaaaatcgat ttgaggatgt ataaagctag catggtgagc aagggcgagg aggataacat
      301 ggccatcatc aaggagttca tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca
      361 cgagttcgag atcgagggcg agggcgaggg ccgcccctac gagggcaccc agaccgccaa
      421 gctgaaggtg accaagggtg gccccctgcc cttcgcctgg gacatcctgt cccctcagtt
      481 catgtacggc tccaaggcct acgtgaagca ccccgccgac atccccgact acttgaagct
      541 gtccttcccc gagggcttca attgggagcg cgtgatgaac ttcgaggacg gcggcgtggt
      601 gaccgtgacc caggactcct ccctgcagga cggcgagttc atctacaagg tgaagctgcg
      661 cggcaccaac ttcccctccg acggccccgt aatgcagtgt cgtaccatgg gctgggaggc
      721 ctccactgag cggatgtacc ccgaggacgg cgccctgaag ggcgagatca agcagaggct
      781 gaagctgaag gacggcggcc actacgacgc tgaggtcaag accacctaca aggccaagaa
      841 gcccgtgcag ctgcccggcg cctacaacgt cgacatcaag ttggacatcc tttcccacaa
      901 cgaggactac accatcgtgg aacagtacga acgcgccgag ggccgccact ccaccggcgg
      961 catggacgag ctgtacaagt aatctagata actgatcata atcagccata ccacatttgt
     1021 agaggtttta cttgctttaa aaaacctccc acacctcccc
//
