LOCUS       sequence-138245-        1102 bp    DNA     linear   UNA 14-APR-2025
DEFINITION  C-terminal tagging with mAID using CRISPR/Cas9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1102)
  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 1102)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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: pMK288 (mAID-Bsr)
FEATURES             Location/Qualifiers
     source          1..1102
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     91..113
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     105..122
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     106..122
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     132..151
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        132..150
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             187..390
                     /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    511..632
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(548..567)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     602..621
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    663..696
                     /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(792..815)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
ORIGIN
        1 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
       61 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
      121 gtgagcgcgc gtaatacgac tcactatagg gcgaattgga gctccccggg atccggtgca
      181 ggcgccaagg agaagagtgc ttgtcctaaa gatccagcca aacctccggc caaggcacaa
      241 gttgtgggat ggccaccggt gagatcatac cggaagaacg tgatggtttc ctgccaaaaa
      301 tcaagcggtg gcccggaggc ggcggcgttc gtgaaggtat caatggacgg agcaccgtac
      361 ttgaggaaaa tcgatttgag gatgtataaa taatctagat aactgatcat aatcagccat
      421 accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg
      481 aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac
      541 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
      601 tgtggtttgt ccaaactcat caatgtatct taacgcgtcg atcatattca ataaccctta
      661 atataacttc gtataatgta tgctatacga agttattagg tctgaagagg agtttacgtc
      721 cagccaagct taggatctcg acctcgaaat tctaccgggt aggggaggcg cttttcccaa
      781 ggcagtctgg agcatgcgct ttagcagccc cgctgggcac ttggcgctac acaagtggcc
      841 tctggcctcg cacacattcc acatccaccg gtaggcgcca accgactccg ttctttggtg
      901 gccccttcgc gccaccttct actcctcccc tagtcaggaa gttccccccc gccccgcagc
      961 tcgcgtcgtg caggacgtga caaatggaag tagcacgtct cactagtctc gtgcagatgg
     1021 acagcaccgc tgagcaatgg aagcgggtag gcctttgggg cagcggccaa tagcagcttt
     1081 gctccttcgc tttctgggct ca
//
