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pMK292 (mAID-mCherry2-NeoR)

A plasmid for construction of C-terminal mAID-mCherry tagging donors.

Catalog number RDB13935
Resource name pMK292 (mAID-mCherry2-NeoR)
Alternative name mAID-mCherry2-NeoR, 971
Clone info. A plasmid for construction of C-terminal mAID-mCherry tagging donors. Please refer Fig. 4 of Natsume, T., Cell Reports 15, 210-218 (2016) for an instruction about how to use this clone. Please note that this plasmid does not contain the loxP sites, which are intended for removal of the marker.
To make 0.5 M stock solution, dissolve indole-3-acetic acid (IAA) (Sigma-Aldrich, 45533) or 1-naphthaleneacetic acid (NAA) (Sigma-Aldrich, 35745) in 100 % DMSO. IAA solution can be made in 100 % ethanol. Aliquots can be kept for 6 months in the dark at -20oC (Devrekanli, A., Kanemaki, M.T. Methods in Molecular Biology, 1369, 257-278, 2016).
Vector backbone pBluescriptII KS- (Plasmid)
Size of vector backbone 3.0 kb
Selectable markers Am^r
Gene/insert name Arabidopsis thaliana AtIAA17 cDNA Discosoma sp. RFP cDNA
Depositor|Developer Kanemaki, Masato |
Other clones in our bank

External Database
Arabidopsis thaliana AtIAA17 Discosoma sp. RFP

          Reference sequence
            

          Distribution information

          Please check terms and conditions set forth by the depositor, which are specified in the RIKEN BRC Catalog and/or Web Catalog.
          Terms and conditions set forth by the DEPOSITOR In publishing the research results obtained by use of the BIOLOGICAL RESOURCE, a citation of the following literature designated by the DEPOSITOR is requested (Natsume, T., Cell Reports, 15: 210-218, 2016). This DNA clone is used for academic research purpose only. This is not to be used on a commercial basis. For use of this DNA clone except for internal academic research, the RECIPIENT must contact a NIG license representative at chizai@nig.ac.jp.
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          The BIOLOGICAL RESOURCE is not provided to for-profit organization or for for-profit research by non-profit organizations. Please read the Exclusive MTA (Takara Bio USA, Inc. FPs, for use for not-for-profit academic purpose).
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          RDB13935 pMK292 (mAID-mCherry2-NeoR) DNA solution

          Ordering Information [in Japanese] [in English]
          Please review the QC test results indicated by check icon below as well as clone information before placing your order.

          References and tips

          Electronic file

          Original reference

          original Natsume, T., Rapid protein depletion in human cells by auxin-inducible degron tagging with short homology donors. Cell Rep. 15 (1): 210-218 (2016). PMID 27052166.

          Further references such as user reports and related articles (go to bottom)

          Featured content

          Featured content Auxin Inducible Degron (AID) System (English text)
          Featured content Auxin Inducible Degron (AID) System (Japanese text)

          Sequence information

          RIKEN BRC has sequenced portions of this material for quality test.
          Please review the QC test results indicated by check icon as well as clone information before placing your order.

          Test sheet RDB13935_A6B8p1-3.pdf check

          Nucleotide sequence of a portion of this resource (if available).

          Primer: M13_-40
          Region: insert5'(Linker,mAID,mCherry2)
          Sequence file: RDB13935_A6B8e.seq check
          >D03070A19_A6B8_1_M13(-40)_D05_11.ab1
              1 CCGGGCAGAT GGAGCGCGCG TAATACGACT CACTATAGGG CGAATTGGAG CTCCCCGGGA
             61 TCCGGTGCAG GCGCCAAGGA GAAGAGTGCT TGTCCTAAAG ATCCAGCCAA ACCTCCGGCC
            121 AAGGCACAAG TTGTGGGATG GCCACCGGTG AGATCATACC GGAAGAACGT GATGGTTTCC
            181 TGCCAAAAAT CAAGCGGTGG CCCGGAGGCG GCGGCGTTCG TGAAGGTATC AATGGACGGA
            241 GCACCGTACT TGAGGAAAAT CGATTTGAGG ATGTATAAAG CTAGCATGGT GAGCAAGGGC
            301 GAGGAGGATA ACATGGCCAT CATCAAGGAG TTCATGCGCT TCAAGGTGCA CATGGAGGGC
            361 TCCGTGAACG GCCACGAGTT CGAGATCGAG GGCGAGGGCG AGGGCCGCCC CTACGAGGGC
            421 ACCCAGACCG CCAAGCTGAA GGTGACCAAG GGTGGCCCCC TGCCCTTCGC CTGGGACATC
            481 CTGTCCCCTC AGTTCATGTA CGGCTCCAAG GCCTACGTGA AGCACCCCGC CGACATCCCC
            541 GACTACTTGA AGCTGTCCTT CCCCGAGGGC TTCAATTGGG AGCGCGTGAT GAACTTCGAG
            601 GACGGCGGCG TGGTGACCGT GACCCAGGAC TCCTCCCTGC AGGACGGCGA GTTCATCTAC
            661 AAGGTGAAGC TGCGCGGCAC CAACTTCCCC TCCGACGGCC CCGTAATGCA GTGTCGTACC
            721 ATGGGCTGGG AGGCCTCCAC TGAGCGGATG TAACCCCGAG GACGGCGCCC TGAAGGGCGA
            781 GATCAAGCAG AGGCTGAAAG CTGAAGGACG GCGGCCACTA CGACGCTGAG GTCAAGACCA
            841 CCTACAAGGC CAAGAAGCCC GTGCAGCTGC CCGGCGCCTA CAACGTCGAC ATCAAGTTGG
            901 GACATCCTTT CCCACAACGA GGACTACACC ATCGTGGAAA CAGTACGAAA CGCGCCGAGG
            961 GGCCGCCCAC TCCACCGGCC GGCATGGAAC GAGCTGTACA AGTAATCTAA GATAAACTGA
           1021 TACATAAATC AGCCATACCA CATTTGGTAA GAGGGTTTTT ACTATGCTTT TAAAAAACTT
           1081 CCCACACCCT TCCCCCTTGA ACTGAAACCA TAGAATGAAT GCAATTGATT GTGGGTAACT
           1141 TGGTTTTATT GGCAAGGCCT TATCAACGTG GGTTTGTAGC AGACAATTGA AGAGCCGTCA
          1201 T
          //
          Primer: SV40polyA-R
          Region: insert mid(SV40 ter,mCherry2)
          Sequence file: RDB13935_A6B8f.seq check
          >D03070A19_A6B8_1_SV40polyA-R_H05_23.ab1
              1 CGCCTCGCAG TTATTAGCAG TAACCTCTAC AATGTGGTAT GGCTGATTAT GATCAGTTAT
             61 CTAGATTACT TGTACAGCTC GTCCATGCCG CCGGTGGAGT GGCGGCCCTC GGCGCGTTCG
            121 TACTGTTCCA CGATGGTGTA GTCCTCGTTG TGGGAAAGGA TGTCCAACTT GATGTCGACG
            181 TTGTAGGCGC CGGGCAGCTG CACGGGCTTC TTGGCCTTGT AGGTGGTCTT GACCTCAGCG
            241 TCGTAGTGGC CGCCGTCCTT CAGCTTCAGC CTCTGCTTGA TCTCGCCCTT CAGGGCGCCG
            301 TCCTCGGGGT ACATCCGCTC AGTGGAGGCC TCCCAGCCCA TGGTACGACA CTGCATTACG
            361 GGGCCGTCGG AGGGGAAGTT GGTGCCGCGC AGCTTCACCT TGTAGATGAA CTCGCCGTCC
            421 TGCAGGGAGG AGTCCTGGGT CACGGTCACC ACGCCGCCGT CCTCGAAGTT CATCACGCGC
            481 TCCCAATTGA AGCCCTCGGG GAAGGACAGC TTCAAGTAGT CGGGGATGTC GGCGGGGTGC
            541 TTCACGTAGG CCTTGGAGCC GTACATGAAC TGAGGGGACA GGATGTCCCA GGCGAAGGGC
            601 AGGGGGCCAC CCTTGGTCAC CTTCAGCTTG GCGGTCTGGG TGCCCTCGTA GGGGCGGCCC
            661 TCGCCCTCGC CCTCGATCTC GAACTCGTGG CCGTTCACGG AGCCCTCCAT GTGCACCTTG
            721 AAGCGCATGA ACTCCTTGAT GATGGCCATG TTATCCTCCT CGCCCTTGCT CACCATGCTA
            781 GCTTTATACA TCCTCAAATC GATTTTCCTC AAGTACGGTG CTCCGTCCAT TGATACCTTC
            841 ACGAACGCCG CCCGCCTCCG GGCCACCGCT TGATTTTTGG CAGGAAACCA TCACGTTCTT
            901 CCGGTATGAT CTCACCGGTG GCCATCCCAC AACTTGTGCC CTTGGCCGGA AGGTTTGGGC
            961 TGGGATCTTT AGGAACAAGC AACTCCTTCT CCCTTGGCGC CTGCACCGGA ATCCCGGGGG
           1021 GAGCTCCAAT TCGCCCTATA AGTGAGTCGT AATACGCGCG CTCCCACTGG CGGTCGATTT
           1081 ACCACGTCCT GACTGGAAAC CCTGCGGTTA CCCACTTAAA TCGCCCTTGG CAGCCACAAT
          1141 CCCCCATATC AGCCACAAGC T
          //
          Primer: SV40pro_F_V2
          Region: insert mid(NeoR)
          Sequence file: RDB13935_A6B8g.seq check
          >D03070A19_A6B8_1_SV40pro_F_V2_F10_16.ab1
              1 GTTGGCCCTT TTTTGGAGGC TAGGCTTTTG CAAGATCGAT CAAGAGACAG GATGAGGATC
             61 GTTTCGCATG ATTGAACAAG ATGGATTGCA CGCAGGTTCT CCGGCCGCTT GGGTGGAGAG
            121 GCTATTCGGC TATGACTGGG CACAACAGAC AATCGGCTGC TCTGATGCCG CCGTGTTCCG
            181 GCTGTCAGCG CAGGGGCGCC CGGTTCTTTT TGTCAAGACC GACCTGTCCG GTGCCCTGAA
            241 TGAACTGCAA GACGAGGCAG CGCGGCTATC GTGGCTGGCC ACGACGGGCG TTCCTTGCGC
            301 AGCTGTGCTC GACGTTGTCA CTGAAGCGGG AAGGGACTGG CTGCTATTGG GCGAAGTGCC
            361 GGGGCAGGAT CTCCTGTCAT CTCACCTTGC TCCTGCCGAG AAAGTATCCA TCATGGCTGA
            421 TGCAATGCGG CGGCTGCATA CGCTTGATCC GGCTACCTGC CCATTCGACC ACCAAGCGAA
            481 ACATCGCATC GAGCGAGCAC GTACTCGGAT GGAAGCCGGT CTTGTCGATC AGGATGATCT
            541 GGACGAAGAG CATCAGGGGC TCGCGCCAGC CGAACTGTTC GCCAGGCTCA AGGCGAGCAT
            601 GCCCGACGGC GAGGATCTCG TCGTGACCCA TGGCGATGCC TGCTTGCCGA ATATCATGGT
            661 GGAAAATGGC CGCTTTTCTG GATTCATCGA CTGTGGCCGG CTGGGTGTGG CGGACCGCTA
            721 TCAGGACATA GCGTTGGCTA CCCGTGATAT TGCTGAAGAG CTTGGCGGCG AATGGGCTGA
            781 CCGCTTCCTC GTGCTTTACG GTATCGCCGC TCCCGATTCG CAGCGCATCG CCTTCTATCG
            841 CCTTCTTGAC GAGTTCTTCT GAGCGGGACT CTGGGGTTCG AAATGACCGA CCAAGCGACG
            901 CCCAACCTGC CATCACGAGA TTTCGATTCC ACCGCCGCCT TCTATGAAAG GTTGGGGCTT
            961 CGGAATCGTT TTCCCGGGAC GCCGGCTGGA TGATCCTCCA GCGCGGGGAT CTCATGCTGG
           1021 GAGTTCTTTC GCCCACCCTA GGGGGAAGGC TAACTGAAAT CACGAAGGAG ACAATACCGG
          1081 AAGGACCGCG CTATGAACCG GCAATTAAAA AGACAGGAAT AAAAACGGCA
          //
          Primer: M13R
          Region: insert3'(NeoR)
          Sequence file: RDB13935_A6B8h.seq check
          >D03070A19_A6B8_1_M13R_D04_10.ab1
              1 GCCGCGGCAT TACCTCACTA AAGGGAACAA AAGCTGGGTA CCGGATCCAA CGACCCAACA
             61 CCGTGCGTTT TATTCTGTCT TTTTATTGCC GTCATAGCGC GGGTTCCTTC CGGTATTGTC
            121 TCCTTCCGTG TTTCAGTTAG CCTCCCCCTA GGGTGGGCGA AGAACTCCAG CATGAGATCC
            181 CCGCGCTGGA GGATCATCCA GCCGGCGTCC CGGAAAACGA TTCCGAAGCC CAACCTTTCA
            241 TAGAAGGCGG CGGTGGAATC GAAATCTCGT GATGGCAGGT TGGGCGTCGC TTGGTCGGTC
            301 ATTTCGAACC CCAGAGTCCC GCTCAGAAGA ACTCGTCAAG AAGGCGATAG AAGGCGATGC
            361 GCTGCGAATC GGGAGCGGCG ATACCGTAAA GCACGAGGAA GCGGTCAGCC CATTCGCCGC
            421 CAAGCTCTTC AGCAATATCA CGGGTAGCCA ACGCTATGTC CTGATAGCGG TCCGCCACAC
            481 CCAGCCGGCC ACAGTCGATG AATCCAGAAA AGCGGCCATT TTCCACCATG ATATTCGGCA
            541 AGCAGGCATC GCCATGGGTC ACGACGAGAT CCTCGCCGTC GGGCATGCTC GCCTTGAGCC
            601 TGGCGAACAG TTCGGCTGGC GCGAGCCCCT GATGCTCTTC GTCCAGATCA TCCTGATCGA
            661 CAAGACCGGC TTCCATCCGA GTACGTGCTC GCTCGATGCG ATGTTTCGCT TGGTGGTCGA
            721 ATGGGCAGGT AGCCGGATCA AGCGTATGCA GCCGCCGCAT TGCATCAGCC ATGATGGATA
            781 CTTTCTCGGC AGGAGCAAGG TGAGATGACA GGAGATCCTG CCCCGGCACT TCGCCCAATA
            841 GCAGCCAGTC CCTTCCCGCT TCAGTGACAA CGTCGAGCAC AGCTGCGCAA GGAACGCCCG
            901 TCGTGGCCAG CCACGATAGC CGCGCTGCCT CGTCTTGCAG TTCATTCAGG GCACCGGACA
            961 GGTCGGTCTT GACAAAAAGA ACCGGGCGCC CCTGCGCTGA CAGCCCGGAA CACGCGGCAT
           1021 CAGAGCAGCC CGATTGTCTG TGTGCCCAGT CATAGGCCGG ATAGGCCTCT CCACCCCAAG
           1081 GCGGGCCAGG AGATCTTGCG ATGCCAGTTC CAATTCTTGA TCAGTCATGC CGAAAACCGA
          1141 TTCCCT
          //

          Please visit Sequencing and PCR primers for primer information.


          References

          Original, user report and related articles

          original Natsume, T., Rapid protein depletion in human cells by auxin-inducible degron tagging with short homology donors. Cell Rep. 15 (1): 210-218 (2016). PMID 27052166.
          reference Goto, H., Chk1-mediated Cdc25A degradation as a critical mechanism for normal cell cycle progression. J Cell Sci. 2019 Jan 25;132(2). pii: jcs223123 (2019). PMID 30635443.
          reference Devrekanli, A., Conditional Budding Yeast Mutants with Temperature-Sensitive and Auxin-Inducible Degrons for Screening of Suppressor Genes. Methods in Molecular Biology 1369: 257-278 (2016). PMID 26519318.
          reference Nishimura, K., Rapid Depletion of Budding Yeast Proteins via the Fusion of an Auxin-Inducible Degron (AID). Curr. Protoc. Cell Biol. \64: 20.9.1-16 (2014). PMID 25181302.

          2021.11.02

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