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Please review the QC test results indicated by check icon below as well as clone information before placing your order.

pMK290 (mAID-mClover-Hygro)

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

Catalog number RDB13933
Resource name pMK290 (mAID-mClover-Hygro)
Alternative name mAID-mClover-Hygro, 1025
Clone info. A plasmid for construction of C-terminal mAID-mClover tagging donors. Please refer Fig. 4 of Natsume, T., Cell Reports 15, 210-218 (2016) for an instruction about how to use this clone.
Comment 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 Aequorea victoria GFP cDNA
Depositor|Developer Kanemaki, Masato |
Other clones in our bank

External Database
Arabidopsis thaliana AtIAA17 Aequorea victoria GFP

          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 for distribution 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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          提供条件 利用者は、研究成果の公表にあたって寄託者の指定する文献を引用する (Natsume, T., Cell Reports, 15: 210-218, 2016)。学術研究にのみ限定、商業営利目的利用を禁ずる。学術内部利用以外の利用の際は、国立遺伝学研究所 (chizai@nig.ac.jp) に相談すること。
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          手続きの概要は、「提供申込みについて[link]」をご覧ください。

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          RDB13933 pMK290 (mAID-mClover-Hygro) DNA solution

          Please review the QC test results indicated by check icon below as well as clone information before placing your order.

          How to cite this biological resource

          Materials & Methods section:

          The pMK290 (mAID-mClover-Hygro) was provided by the RIKEN BRC through the National BioResource Project of the MEXT, Japan (cat. RDB13933).

          Reference section:

          Natsume, T., Kiyomitsu, T., Saga, Y., Kanemaki, M.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. [link to RRC of NBRP]

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


          Tips

          Featured content

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

          QC test results

          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 RDB13933_A6B8p1-3.pdf check

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

          Primer: M13_-40 (Pr0410)
          Region: insert mid(Linker,mAID,mClover(A206K) 5')
          Sequence file: RDB13933_A6B8i.seq check
          >D03070A17_A6B8_1_M13(-40)_B05_05.ab1
              1 NNNNNNNNNN NNNNCNNANA NNNNTNNNNA TAGGGCGAAN TGGAGCTCCC CGGGNTCCGG
             61 TGCAGGCGCC AAGGAGAAGA GTGCTTGTCC TAAAGATCCA GCCAAACCTC CGGCCAAGGC
            121 ACAAGTTGTG GGATGGCCAC CGGTGAGATC ATACCGGAAG AACGTGATGG TTTCCTGCCA
            181 AAAATCAAGC GGTGGCCCGG AGGCGGCGGC GTTCGTGAAG GTATCAATGG ACGGAGCACC
            241 GTACTTGAGG AAAATCGATT TGAGGATGTA TAAAGCTAGC ATGGTGAGCA AGGGCGAGGA
            301 GCTGTTCACC GGGGTGGTGC CCATCCTGGT CGAGCTGGAC GGCGACGTAA ACGGCCACAA
            361 GTTCAGCGTC CGCGGCGAGG GCGAGGGCGA TGCCACCAAC GGCAAGCTGA CCCTGAAGTT
            421 CATCTGCACC ACCGGCAAGC TGCCCGTGCC CTGGCCCACC CTCGTGACCA CCTTCGGCTA
            481 CGGCGTGGCC TGCTTCAGCC GCTACCCCGA CCACATGAAG CAGCACGACT TCTTCAAGTC
            541 CGCCATGCCC GAAGGCTACG TCCAGGAGCG CACCATCTCT TTCAAGGACG ACGGTACCTA
            601 CAAGACCCGC GCCGAGGTGA AGTTCGAGGG CGACACCCTG GTGAACCGCA TCGAGCTGAA
            661 GGGCATCGAC TTCAAGGAGG ACGGCAACAT CCTGGGGCAC AAGCTGGAGT ACAACTTCAA
            721 CAGCCACAAC GTCTATATCA CGGCCGACAA GCAGANNAAC GGCATCNAGG CTAACTTCAA
            781 GATCCGNCAC NACGTTGAGG ACGGCAGCNN GCANCTNNGC CGACCACNAC NAGCANANNA
            841 CCCCCATCGG CGACGGNCCC GNGCTGCTGC NNNACNACNN NTACCTNANC CNTCAGTNNN
            901 AGCTGAGCAA ANACCCCANN NANNANCGCN NNTNANNNNG NNNCNTGCNN GNNNTTCGTG
          961 ACNNNCNNNC NGGNNNNNNN NNN
          //
          Primer: M13R (Pr0160)
          Region: insert 3'(loxP,mPGK ter,HygR 3')
          Sequence file: RDB13933_A6B8j.seq check
          >D03070A17_A6B8_1_M13R_B04_04.ab1
              1 NNNNNNNNNN NNNNNNNNNN NNAAGGGNNN AAAAGCTGGG TACCGGATCC TAGTGAACCT
             61 CTTCGAGGGA CCTAATAACT TCGTATAGCA TACATTATAC GAAGTTATAT TAAGGGTTCC
            121 GGATCTCGAC CAGCTTCTGA TGGAATTAGA ACTTGGCAAA ACAATACTGA GAATGAAGTG
            181 TATGTGGAAC AGAGGCTGCT GATCTCGTTC TTCAGGCTAT GAAACTGACA CATTTGGAAA
            241 CCACAGTACT TAGAACCACA AAGTGGGAAT CAAGAGAAAA ACAATGATCC CACGAGAGAT
            301 CTATAGATCT ATAGATCATG AGTGGGAGGA ATGAGCTGGC CCTTAATTTG GTTTTGCTTG
            361 TTTAAATTAT GATATCCAAC TATGAAACAT TATCATAAAG CAATAGTAAA GAGCCTTCAG
            421 TAAAGAGCAG GCATTTATCT AATCCCACCC CACCCCCACC CCCGTAGCTC CAATCCTTCC
            481 ATTCAAAATG TAGGTACTCT GTTCTCACCC TTCTTAACAA AGTATGACAG GAAAAACTTC
            541 CATTTTAGTG GACATCTTTA TTGTTTAATA GATCATCAAT TTCTGCATCC CGGGGATCTG
            601 ATATCATCGA TGCATGGGGT CGTGCGCTCC TTTCGGTCGG GCGCTGCGGG TCGTGGGGCG
            661 GGCGGCTAGC CTATTCCTTT GCCCTCGGAC GAGTGCTGGG GCGTCGGTTT CCACTATCGG
            721 CGAGTACTTC TACACAGCCA TCGGTCCAGA CNGCCGCGCT TCTGCGGGCG ANTTGTGTAC
            781 GCCCGACAGT CCCGGCTCCG GATCGGANGA TTGCGTCGCA TCGACCCTGC GCCCNAGCTG
            841 CATCATCNAA ATTGCCGTCN ACCNAGCTCT GANAGANTTN NTCAAGANCA ATGCGGANCA
            901 TATACNCCNG GANCCNCGGC NNTCCTGNNA GCNNNNGANG CCNNCGCTCN NAGTNNNNNG
          961 NNTGNNGCNN NNNANNNGCC NANCNNNGGN CNNNNNNNNN N
          //
          Primer: PGK_N_rev (Pr0243)
          Region: insert mid(loxP,SV40 pA,mClover(A206K) 3')
          Sequence file: RDB13933_A6B8k.seq check
          >D03070A17_A6B8_1_PGK-N-rev_E06_15.ab1
              1 NNNNNNNNNN NNNNNCNNNN NNNNANNNNT TCNAGGTCGA GATCCTAAGC TTGGCTGGAC
             61 GTAAACTCCT CTTCAGACCT AATAACTTCG TATAGCATAC ATTATACGAA GTTATATTAA
            121 GGGTTATTGA ATATGATCGA CGCGTTAAGA TACATTGATG AGTTTGGACA AACCACAACT
            181 AGAATGCAGT GAAAAAAATG CTTTATTTGT GAAATTTGTG ATGCTATTGC TTTATTTGTA
            241 ACCATTATAA GCTGCAATAA ACAAGTTAAC AACAACAATT GCATTCATTT TATGTTTCAG
            301 GTTCAGGGGG AGGTGTGGGA GGTTTTTTAA AGCAAGTAAA ACCTCTACAA ATGTGGTATG
            361 GCTGATTATG ATCAGTTATC TAGATTACTT GTACAGCTCG TCCATGCCAT GTGTAATCCC
            421 GGCGGCGGTC ACGAACTCCA GCAGGACCAT GTGATCGCGC TTCTCGTTGG GGTCTTTGCT
            481 CANCTTGGAC TGATGGCTCA GGTAGTGGTT GTCGGGCAGC AGCACGGGGC CGTCGCCGAT
            541 GGGGGTGTTC TGCTGGTAGT GGTCGGCGAG CTGCACGCNN NNNNCCTCAA CGTTGTGGCG
          601 GATCTTGAAG TTANNNNNNN NNN
          //
          Primer: PGKpro_F (Pr0249)
          Region: HygR 5'
          Sequence file: RDB13933_A6B8l.seq check
          >D03070A17_A6B8_1_PGK-proF_H07_22.ab1
              1 NNNNNNNNNN NNNNNNNNNN NTCTCCNGGC CTTTCGACCT GCATCCNTCT AGATCTCGAG
             61 CAGCTGAAGC TTACCGCTAG CATGGATAGA TCCGGAAAGC CTGAACTCAC CGCGACGTCT
            121 GTCGAGAAGT TTCTGATCGA AAAGTTCGAC AGCGTCTCCG ACCTGATGCA GCTCTCGGAG
            181 GGCGAAGAAT CTCGTGCTTT CAGCTTCGAT GTAGGAGGGC GTGGATATGT CCTGCGGGTA
            241 AATAGCTGCG CCGATGGTTT CTACAAAGAT CGTTATGTTT ATCGGCACTT TGCATCGGCC
            301 GCGCTCCCGA TTCCGGAAGT GCTTGACATT GGGGAATTCA GCGAGAGCCT GACCTATTGC
            361 ATCTCCCGCC GTGCACAGGG TGTCACGTTG CAAGACCTGC CTGAAACCGA ACTGCCCGCT
            421 GTTCTGCAGC CGGTCGCGGA GGCCATGGAT GCGATCGCTG CGGCCGATCT TAGCCAGACG
            481 AGCGGGTTCG GCCCATTCGG ACCGCAAGGA ATCGGTCAAT ACACTACATG GCGTGATTTC
            541 ATATGCGCGA TTGCTGATCC CCATGTGTAT CACTGGCAAA CTGTGATGGA CGACACCGTC
            601 AGTGCGTCCG TCGCGCAGGC TCTCGATGAG CTGATGCTTT GGGCCGAGGA CTGCCCCGAA
            661 GTCCGGCACC TCGTGCACGC GGATTTCGGC TCCNANAATG TCCTGACGGA CAATGGCCGC
            721 ATAACAGCGG TCATTGACTG GAGCGAGGCG ATGTTCGGGG ATTCCCAATA CGAGGTCGCC
            781 AACATCTTCT TCTGGANGCC GTGGTTGGCT TGTATGGAGC AGCANACGCG CTACTTCGNG
            841 CGGANGCNTC CGGAGCTNGC NNNTCGCCGC NGCTCCNNNC GTATATGCTC CGCNTTNNNC
            901 NTGANCAACT CTATCANAGC TNGNNTNGAN NNCNNTTTNN ATGANGCANC TNNNGNGCNN
            961 NNTCGANGNN ACGCNNNCNN NCCNNNNCNG ANNCNNGNNN NGNNNNGGNG NNNNNNANNN
          1021 NNNNNNN
          //
          Primer: RV5F (pGL3_RV5F) (Pr0227)
          Region: insert mid(mClover(A206K) mid,mAID 3')
          Sequence file: RDB13933_A6B8m.seq check
          >13933_13933_B2JhF_RV5F_C02_03_ABI08.ab1
              1 NNNNNNNNNN NNNNNGNNNN NGGNTGNNTN NGNNNNGTTA TCTAGATTAC TTGTACAGCT
             61 CGTCCATGCC ATGTGTAATC CCGGCGGCGG TCACGAACTC CAGCAGGACC ATGTGATCGC
            121 GCTTCTCGTT GGGGTCTTTG CTCAGCTTGG ACTGATGGCT CAGGTAGTGG TTGTCGGGCA
            181 GCAGCACGGG GCCGTCGCCG ATGGGGGTGT TCTGCTGGTA GTGGTCGGCG AGCTGCACGC
            241 TGCCGTCCTC AACGTTGTGG CGGATCTTGA AGTTAGCCTT GATGCCGTTC TTCTGCTTGT
            301 CGGCCGTGAT ATAGACGTTG TGGCTGTTGA AGTTGTACTC CAGCTTGTGC CCCAGGATGT
            361 TGCCGTCCTC CTTGAAGTCG ATGCCCTTCA GCTCGATGCG GTTCACCAGG GTGTCGCCCT
            421 CGAACTTCAC CTCGGCGCGG GTCTTGTAGG TACCGTCGTC CTTGAAAGAG ATGGTGCGCT
            481 CCTGGACGTA GCCTTCGGGC ATGGCGGACT TGAAGAAGTC GTGCTGCTTC ATGTGGTCGG
            541 GGTAGCGGCT GAAGCAGGCC ACGCCGTAGC CGAAGGTGGT CACGAGGGTG GGCCAGGGCA
            601 CGGGCAGCTT GCCGGTGGTG CAGATGAACT TCAGGGTCAG CTTGCCGTTG GTGGCATCGC
            661 CCTCGCCCTC GCCGCGGACG CTGAACTTGT GGCCGTTTAC GTCGCCGTCC AGCTCGACCA
            721 GGATGGGCAC CACCCCGGTG AACAGCTCCT CGCCCTTGCT CACCATGCTA GCTTTATACA
            781 TCCTCAAATC GATTTTCCTC AAGTACGGTG CTCCGTCCAT TGATACCTTC ACGAACGCCG
            841 CCGCCTCCNG NNCANCGCTT GANTTTTGGC NNNAACCATC ACGTTCNTNC CNGTATGATC
            901 TCACNNGNGG NCATCCCACA NCTTGTGCCT TGGCCNGNNG NTTTGGCNNG NATCTTTNNN
          961 NNAGCACTCT TNNCCTNNGN CNNNNNNNNN CNGNNTNCNN GNNNNNNNNN N
          //

          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. [link to RRC of NBRP]
          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 Takagi, M., Perichromosomal protein Ki67 supports mitotic chromosome architecture. Genes Cells 21 (10): 1113-1124 (2016). PMID 27610954.
          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.

          2023.10.05

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