LOCUS       sequence-138280-        1116 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 1116)
  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 1116)
  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: pMK293 (mAID-mCherry2-Hygro)
FEATURES             Location/Qualifiers
     source          1..1116
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             47..250
                     /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             257..967
                     /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(402..420)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
ORIGIN
        1 tcactatagg gcgaattgga gctccccggg atccggtgca ggcgccaagg agaagagtgc
       61 ttgtcctaaa gatccagcca aacctccggc caaggcacaa gttgtgggat ggccaccggt
      121 gagatcatac cggaagaacg tgatggtttc ctgccaaaaa tcaagcggtg gcccggaggc
      181 ggcggcgttc gtgaaggtat caatggacgg agcaccgtac ttgaggaaaa tcgatttgag
      241 gatgtataaa gctagcatgg tgagcaaggg cgaggaggat aacatggcca tcatcaagga
      301 gttcatgcgc ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt tcgagatcga
      361 gggcgagggc gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa
      421 gggtggcccc ctgcccttcg cctgggacat cctgtcccct cagttcatgt acggctccaa
      481 ggcctacgtg aagcaccccg ccgacatccc cgactacttg aagctgtcct tccccgaggg
      541 cttcaattgg gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg tgacccagga
      601 ctcctccctg caggacggcg agttcatcta caaggtgaag ctgcgcggca ccaacttccc
      661 ctccgacggc cccgtaatgc agtgtcgtac catgggctgg gaggcctcca ctgagcggat
      721 gtaccccgag gacggcgccc tgaagggcga gatcaagcag aggctgaagc tgaaggacgg
      781 cggccactac gacgctgagg tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc
      841 cggcgcctac aacgtcgaca tcaagttgga catcctttcc cacaacgagg actacaccat
      901 cgtggaacag tacgaacgcg ccgagggccg ccactccacc ggcggcatgg acgagctgta
      961 caagtaatct agataactga tcataatcag ccataccaca tttgtagagg ttttacttgc
     1021 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt
     1081 tgttaacttg tttattgcag cttataatgg ttacaa
//
