LOCUS       sequence-138166-         979 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  C-terminal tagging with mAID using CRISPR/Cas9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 979)
  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 979)
  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: pMK287 (mAID-Hygro)
FEATURES             Location/Qualifiers
     source          1..979
                     /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"
     polyA_signal    386..507
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(423..442)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     477..496
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    538..571
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(667..690)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, 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 ataaataatc tagataactg atcataatca gccataccac
      301 atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca
      361 taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata
      421 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
      481 tttgtccaaa ctcatcaatg tatcttaacg cgtcgatcat attcaataac ccttaatata
      541 acttcgtata atgtatgcta tacgaagtta ttaggtctga agaggagttt acgtccagcc
      601 aagcttagga tctcgacctc gaaattctac cgggtagggg aggcgctttt cccaaggcag
      661 tctggagcat gcgctttagc agccccgctg ggcacttggc gctacacaag tggcctctgg
      721 cctcgcacac attccacatc caccggtagg cgccaaccga ctccgttctt tggtggcccc
      781 ttcgcgccac cttctactcc tcccctagtc aggaagttcc cccccgcccc gcagctcgcg
      841 tcgtgcagga cgtgacaaat ggaagtagca cgtctcacta gtctcgtgca gatggacagc
      901 accgctgagc aatggaagcg ggtaggcctt tggggcagcg gccaatagca gctttgctcc
      961 ttcgctttct gggctcaga
//
