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

pMK294 (mAID-mCherry2-Bsr)

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

Catalog number RDB13937
Resource name pMK294 (mAID-mCherry2-Bsr)
Alternative name mAID-mCherry2-Bsr, 976
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.
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 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 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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          RDB13937 pMK294 (mAID-mCherry2-Bsr) 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 pMK294 (mAID-mCherry2-Bsr) was provided by the RIKEN BRC through the National BioResource Project of the MEXT, Japan (cat. RDB13937).

          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 RDB13937_A6B8p1-3.pdf check

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

          Primer: M13_-40 (Pr0410)
          Region: insert mid(Linker,mAID,mCherry2 5')
          Sequence file: RDB13937_A6B8e.seq check
          >D03070A21_A6B8_1_M13(-40)_F05_17.ab1
              1 NNNNNNNNNN NNNCNNNNNN GACTNNCNAT AGGGCGAATT GGAGCTCCCC GGGNTCCGGT
             61 GCAGGCGCCA AGGAGAAGAG TGCTTGTCCT AAAGATCCAG CCAAACCTCC GGCCAAGGCA
            121 CAAGTTGTGG GATGGCCACC GGTGAGATCA TACCGGAAGA ACGTGATGGT TTCCTGCCAA
            181 AAATCAAGCG GTGGCCCGGA GGCGGCGGCG TTCGTGAAGG TATCAATGGA CGGAGCACCG
            241 TACTTGAGGA AAATCGATTT GAGGATGTAT AAAGCTAGCA TGGTGAGCAA GGGCGAGGAG
            301 GATAACATGG CCATCATCAA GGAGTTCATG CGCTTCAAGG TGCACATGGA GGGCTCCGTG
            361 AACGGCCACG AGTTCGAGAT CGAGGGCGAG GGCGAGGGCC GCCCCTACGA GGGCACCCAG
            421 ACCGCCAAGC TGAAGGTGAC CAAGGGTGGC CCCCTGCCCT TCGCCTGGGA CATCCTGTCC
            481 CCTCAGTTCA TGTACGGCTC CAAGGCCTAC GTGAAGCACC CCGCCGACAT CCCCGACTAC
            541 TTGAAGCTGT CCTTCCCCGA GGGCTTCAAT TGGGAGCGCG TGATGAACTT CGAGGACGGC
            601 GGCGTGGTGA CCGTGACCCA GGACTCCTCC CTGCAGGACG GCGAGTTCAT CTACAAGGTG
            661 AAGCTGCGCG GCACNNAACT TCCCCTCCGA CGGCCCCGTA ATGNAGTGTC GTACCATGGG
            721 CTGGGANGNC TCCACTGANC GGATGTACCN CGANGANGGC GCCCTGAANG GNGAGNTCAA
            781 NCANANGCTG AAGCTGANNN CGGCGGCCNC TACNANGCTG ANGTCNAGAC CACCTACNAN
            841 GNNNNAGAAG CCCGTGCANC TGCCCNGNGC CNACAANNNC NANNTCANNT TGGANNNCCT
            901 TTNNCCANNN NNNNNACNNN NCCNNNGNNN AACANTNCNA ACGCNNCNNN NGGGNCNNCN
            961 NNTCCNNCNN NNNGNNNNGN NNNNNNCNGT NNNNANNTAN NNNNANANNN NNGNNNNNNN
          1021 ANNNNNNNNN CNNNNNNNNN N
          //
          Primer: M13R (Pr0160)
          Region: insert 3'(loxP,mPGK ter,bsr)
          Sequence file: RDB13937_A6B8f.seq check
          >D03070A21_A6B8_1_M13R_F04_16.ab1
              1 NNNNNNNNNN NNNNNNNNNN NAANGGGNNN NAAAGCTGGG TANCGGATCC 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 TTAATTTCGG GTATATTTGA GTGGAATGAG TTCTTCAATC GTAGTTTTGA
            721 CTAACTTGCC ATTCATTTCT ATTAACACAA AACAATCTGG TGCATAGTCT GAAATCAACT
            781 CCCTACACAT ACCACAAGGA CTTACNNCTC GAATACTTCT ATCTACTTCG TCAGANNANN
            841 NTGTCNNACA GCTACNATCG TGTCNAANTC CTTTTGTNNN TTCNNAACTG CNCNANCNAT
          901 CNCAANGGCN NCTGCNCAAA CNGTTANTCN NNCNNNNNNN NNN
          //
          Primer: PGK_N_rev (Pr0243)
          Region: insert mid(loxP,SV40 pA,mCherry2 3')
          Sequence file: RDB13937_A6B8g.seq check
          >D03070A21_A6B8_1_PGK-N-rev_H06_24.ab1
              1 NNNNNNNNNN NNNNNNNGNN NANTTCGAGG TCGAGNTCCT AAGCTTGGCT GGACGTAAAC
             61 TCCTCTTCNG ACCTAATAAC TTCGTATAGC ATACATTATA CGAAGTTATA TTAAGGGTTA
            121 TTGAATATGA TCGACGCGTT AAGATACATT GATGAGTTTG GACAAACCAC AACTAGAATG
            181 CAGTGAAAAA AATGCTTTAT TTGTGAAATT TGTGATGCTA TTGCTTTATT TGTAACCATT
            241 ATAAGCTGCA ATAAACAAGT TAACAACAAC AATTGCATTC ATTTTATGTT TCAGGTTCAG
            301 GGGGAGGTGT GGGAGGTTTT TTAAAGCAAG TAAAACCTCT ACAAATGTGG TATGGCTGAT
            361 TATGATCAGT TATCTAGATT ACTTGTACAG CTCGTCCATG CCGCCGGTGG AGTGGCGGCC
            421 CTCGGCGCGT TCGTACTGTT CCACGATGGT GTAGTCCTCG TTGTGGGAAA GGATGTCCAA
            481 CTTGATGTCG ACGTTGTAGG CGCCGGGCAG CTGCACGGGC TTCTTGGCCT TGTAGGTGGT
            541 CTTGACCTCA GCGTCGTAGT GGCCGCCGTC CTTCAGCTTC AGCCTCTGCT TGATCTCGCC
            601 CTTCAGGGCG CCGTCCTCGG GGTACATCCG CTCAGTGGAG GCCTCCCAGC CCATGGTACG
            661 ACACTGCATT ACGGGGCCGT CGGANGGGAA GTTGGTGCCG CGCAGCTTCA CCTTGTAGAT
            721 GAACTCGCCG TCCTGCANGG ANGANTCCTG GGTCACGGTC ACCACGCCGC CNTCCTCGAA
            781 NTTCATCACG CGCTCCCAAT TGAAGCCCTC GGGGAANGNN AGCTTCAAGT ANTCNGGGNN
            841 GTCNGCGGGG TGCTTCACGT ANGCCTTNNA NCCNTACATG AACTGANGGG NNAGGANGTC
            901 CNAGGCGAAN GGNANGGGGN CACCCTNNGT NNNCNTCANC TTGGCGGNCN GGGNGCCCNC
          961 NNNNGGCNGN CNNCGCCCTC NNNNNNNTNN NNNACNNNNN NNNNN
          //
          Primer: PGKpro_F (Pr0249)
          Region: insert mid(bsr,mPGK ter)
          Sequence file: RDB13937_A6B8h.seq check
          >D03070A21_A6B8_1_PGK-proF_C08_08.ab1
              1 NNNNNNNNNN NNNNNNNNNN NTCTCCNGGC CTTTCGACCT GCATCCNTCT AGATCTCGAG
             61 CAGCTGAAGC TTACCGCTAG CATGAAAACA TTTAACATTT CTCAACAAGA TCTAGAATTA
            121 GTAGAAGTAG CGACAGAGAA GATTACAATG CTTTATGAGG ATAATAAACA TCATGTGGGA
            181 GCGGCAATTC GTACGAAAAC AGGAGAAATC ATTTCGGCAG TACATATTGA AGCGTATATA
            241 GGACGAGTAA CTGTTTGTGC AGAAGCCATT GCGATTGGTA GTGCAGTTTC GAATGGACAA
            301 AAGGATTTTG ACACGATTGT AGCTGTTAGA CACCCTTATT CTGACGAAGT AGATAGAAGT
            361 ATTCGAGTGG TAAGTCCTTG TGGTATGTGT AGGGAGTTGA TTTCAGACTA TGCACCAGAT
            421 TGTTTTGTGT TAATAGAAAT GAATGGCAAG TTAGTCAAAA CTACGATTGA AGAACTCATT
            481 CCACTCAAAT ATACCCGAAA TTAAGCTAGC CGCCCGCCCC ACGACCCGCA GCGCCCGACC
            541 GAAAGGAGCG CACGACCCCA TGCATCGATG ATATCAGATC CCCGGGATGC AGAAATTGAT
            601 GATCTATTAA ACAATAAAGA TGTCCACTAA AATGGAAGTT TTTCCTGTCA TACTTTGTTA
            661 AGAAGGGTGA GAACAGAGTA CCTACATTTT GAATGGAAGG ATTGGAGCTA CGGGGGTGGG
            721 GGTGGGGTGG GATTAGATAA ATGCCTGCTC TTTACTGAAG GCTCTTTACT ATTGCTTTAT
            781 GATAATGNTT CATAGTTGGA TATCATAATT TAAACAAGCN AAACCAAATT NANGGGNCAG
            841 CTCANTCCTC CCACTCATGA TCNATAGATC TATAGATCTC TCGNGGGATC ATTGNTTTTC
            901 TCTTGANTCC CACTTTNNGG NNCTANGTNN TGNGGTTNNC NNNANGNGNN ANTTTCNNAG
          961 CNNGNANNAN CNAGATNNNN NNNNCNNNNN NNCNNNNNNN NNN
          //
          Primer: mCherry-C_R2 (Pr0792)
          Region: insert mid(mCherry2 5',mAID 3')
          Sequence file: RDB13937_A6B8i.seq check
          >13937_13937_B3C8F_mCherry-C_R2_E01_05_ABI08.ab1
              1 NNNNNNNNNN NNNNCNNNNT TNNGCTTNNG CCTCTGCTTG ATCTCGCCCT TCAGGGCGCC
             61 GTCCTCGGGG TACATCCGCT CAGTGGAGGC CTCCCAGCCC ATGGTACGAC ACTGCATTAC
            121 GGGGCCGTCG GAGGGGAAGT TGGTGCCGCG CAGCTTCACC TTGTAGATGA ACTCGCCGTC
            181 CTGCAGGGAG GAGTCCTGGG TCACGGTCAC CACGCCGCCG TCCTCGAAGT TCATCACGCG
            241 CTCCCAATTG AAGCCCTCGG GGAAGGACAG CTTCAAGTAG TCGGGGATGT CGGCGGGGTG
            301 CTTCACGTAG GCCTTGGAGC CGTACATGAA CTGAGGGGAC AGGATGTCCC AGGCGAAGGG
            361 CAGGGGGCCA CCCTTGGTCA CCTTCAGCTT GGCGGTCTGG GTGCCCTCGT AGGGGCGGCC
            421 CTCGCCCTCG CCCTCGATCT CGAACTCGTG GCCGTTCACG GAGCCCTCCA TGTGCACCTT
            481 GAAGCGCATG AACTCCTTGA TGATGGCCAT GTTATCCTCC TCGCCCTTGC TCACCATGCT
            541 AGCTTTATAC ATCCTCAAAT CGATTTTCCT CAAGTACGGT GCTCCGTCCA TTGATACCTT
            601 CACGAACGCC GCCGCCTCCG GGCCACCGCT TGATTTTTGG CAGGAAACCA TCACGTTCTT
            661 CCGGTATGAT CTCACCGGTG GCCATCCCAC AACTTGTGCC TTGGCCGGAG GTTTGGCTGG
            721 ATCTTTANGA CAAGCACTCT TCTCCTTGGC GCCTGCACCG GATCCCGGGG AGCTCCAATT
            781 CGCCCTATAG TGAGTCGTAT TACGCGCGCT CACTGGCCGT CGTTTTACAA CGTCNTGACT
            841 GNGAAAACCC TGGCGTTACC CAACNTAATC GCCTNNCAGC ACATCCCCCT TTCGCCAGCT
            901 NGCGTAATAN CNAANANGCC CGCACCGATN GCCNNTCCNN NNAGTNGCNC ANCCTGNNTG
          961 GCNNNNNNNA ANTNGNNNGC GNNNNTATTT TNNNNAANNN NNNNNNN
          //

          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 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.

          2023.12.13

          GNP_filter3_RDBDEP_html_231213.pl