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

pMK293 (mAID-mCherry2-Hygro)

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

Catalog number RDB13936
Resource name pMK293 (mAID-mCherry2-Hygro)
Alternative name mAID-mCherry2-Hygro, 977
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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          RDB13936 pMK293 (mAID-mCherry2-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 pMK293 (mAID-mCherry2-Hygro) was provided by the RIKEN BRC through the National BioResource Project of the MEXT, Japan (cat. RDB13936).

          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 RDB13936_A6B8p1-4.pdf check

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

          Primer: M13_-40 (Pr0410)
          Region: T7 pro,insert 5'(Linker,mAID,mCherry2 5')
          Sequence file: RDB13936_A6B8i.seq check
          >D03070A20_A6B8_1_M13(-40)_E05_14.ab1
              1 NNNNNNNNNC NCNNANNNGN NTNNCNATAG GGCGAATTGG AGCTCCCCGG GNTCCGGTGC
             61 AGGCGCCAAG GAGAAGAGTG CTTGTCCTAA AGATCCAGCC AAACCTCCGG CCAAGGCACA
            121 AGTTGTGGGA TGGCCACCGG TGAGATCATA CCGGAAGAAC GTGATGGTTT CCTGCCAAAA
            181 ATCAAGCGGT GGCCCGGAGG CGGCGGCGTT CGTGAAGGTA TCAATGGACG GAGCACCGTA
            241 CTTGAGGAAA ATCGATTTGA GGATGTATAA AGCTAGCATG GTGAGCAAGG GCGAGGAGGA
            301 TAACATGGCC ATCATCAAGG AGTTCATGCG CTTCAAGGTG CACATGGAGG GCTCCGTGAA
            361 CGGCCACGAG TTCGAGATCG AGGGCGAGGG CGAGGGCCGC CCCTACGAGG GCACCCAGAC
            421 CGCCAAGCTG AAGGTGACCA AGGGTGGCCC CCTGCCCTTC GCCTGGGACA TCCTGTCCCC
            481 TCAGTTCATG TACGGCTCCA AGGCCTACGT GAAGCACCCC GCCGACATCC CCGACTACTT
            541 GAAGCTGTCC TTCCCCGAGG GCTTCAATTG GGAGCGCGTG ATGAACTTCG AGGACGGCGG
            601 CGTGGTGACC GTGACCCAGG ACTCCTCCCT GCAGGACGGC GAGTTCATCT ACAAGGTGAA
            661 GCTGCGCGGC ACCAACTTCC CCTCCGACGG CCCCGTAATG CAGTGTCGTA CCATGGGCTG
            721 GGANGCCTCC ACTGAGCGGA TGTACCCCGA GGACGGCGCC CTGAAGGGCG AGATCAAGCA
            781 GANGCTGAAG CTGAANGACG GCGGCCACTA CGACGCTGAN GTCAAGACCA CCTACAAGGC
            841 CNAGAAGCCC GTGCAGCTGC CCGGCGCCTA CNACGTCNAC ATCAAGTTGG ANATCCTTTT
            901 NCCACNACGA NGANNACNNN NTCNNGGNAN CAGTACNAAC GCNCCNAGGG NCGCCNNNNC
            961 NNNNNGNNGG CATGGNNNAN CTGTNNNANT NATCNANNNN NCTGNTCNNA ANCANCCNTA
          1021 NNANNNTNNN NNNNGNNTTT NNNNNNNNNT NANAAANNNN NNNNNN
          //
          Primer: M13R (Pr0160)
          Region: insert 3'(loxP,mouse PGK ter,HygR)
          Sequence file: RDB13936_A6B8j.seq check
          >D03070A20_A6B8_1_M13R_E04_13.ab1
              1 NNNNNNNNNN NNNNNNNNNN ANGNNNNNNA AGCTGGGTAC CGGATCCTAG TGAACCTCTT
             61 CGAGGGACCT AATAACTTCG TATAGCATAC ATTATACGAA GTTATATTAA GGGTTCCGGA
            121 TCTCGACCAG CTTCTGATGG AATTAGAACT TGGCAAAACA ATACTGAGAA TGAAGTGTAT
            181 GTGGAACAGA GGCTGCTGAT CTCGTTCTTC AGGCTATGAA ACTGACACAT TTGGAAACCA
            241 CAGTACTTAG AACCACAAAG TGGGAATCAA GAGAAAAACA ATGATCCCAC GAGAGATCTA
            301 TAGATCTATA GATCATGAGT GGGAGGAATG AGCTGGCCCT TAATTTGGTT TTGCTTGTTT
            361 AAATTATGAT ATCCAACTAT GAAACATTAT CATAAAGCAA TAGTAAAGAG CCTTCAGTAA
            421 AGAGCAGGCA TTTATCTAAT CCCACCCCAC CCCCACCCCC GTAGCTCCAA TCCTTCCATT
            481 CAAAATGTAG GTACTCTGTT CTCACCCTTC TTAACAAAGT ATGACAGGAA AAACTTCCAT
            541 TTTAGTGGAC ATCTTTATTG TTTAATAGAT CATCAATTTC TGCATCCCGG GGATCTGATA
            601 TCATCGATGC ATGGGGTCGT GCGCTCCTTT CGGTCGGGCG CTGCGGGTCG TGGGGCGGGC
            661 GGCTAGCCTA TTCCTTTGCC CTCGGACGAG TGCTGGGGCG TCGGTTTCCA CTATCGGCGA
            721 GTACTTCTAC ACAGCCATCG GTCCAGACGG CCGCGCTTCT GCGGGCGANT TGTGTACGCC
            781 CGACAGTCCC GGCTCCGGAT CGGACGATTG CGTCGCATCN ACCCNGCGCC CAAGCTGCAT
            841 CATCNAAATT GCCGTCNACC AAGCTCTGAT AGAGTTGGTC AAGANNAATG CNNANCATAT
            901 ACGCCNNGAN CCNCGGCGAT NCNGCAANCT CCNGANGNCN NNNCNCNNNN TANCNNGNCN
          961 GCNGNNNNNT ANNNNNCNAN NCNNNNGNNN NNNNNNN
          //
          Primer: PGK_N_rev (Pr0243)
          Region: insert mid(loxP,SV40 pA,mCherry2 3')
          Sequence file: RDB13936_A6B8k.seq check
          >D03070A20_A6B8_1_PGK-N-rev_G06_21.ab1
              1 NNNNNNNNNN NNNNNNNNGN ANANTTCGAG GTCGAGNTCC TAAGCTTGGC TGGACGTAAA
             61 CTCCTCTTCA GACCTAATAA CTTCGTATAG CATACATTAT ACGAAGTTAT ATTAAGGGTT
            121 ATTGAATATG ATCGACGCGT TAAGATACAT TGATGAGTTT GGACAAACCA CAACTAGAAT
            181 GCAGTGAAAA AAATGCTTTA TTTGTGAAAT TTGTGATGCT ATTGCTTTAT TTGTAACCAT
            241 TATAAGCTGC AATAAACAAG TTAACAACAA CAATTGCATT CATTTTATGT TTCAGGTTCA
            301 GGGGGAGGTG TGGGAGGTTT TTTAAAGCAA GTAAAACCTC TACAAATGTG GTATGGCTGA
            361 TTATGATCAG TTATCTAGAT TACTTGTACA GCTCGTCCAT GCCGCCGGTG GAGTGGCGGC
            421 CCTCGGCGCG TTCGTACTGT TCCACGATGG TGTAGTCCTC GTTGTGGGAA AGGATGTCCA
            481 ACTTGATGTC GACGTTGTAG GCGCCGGGCA GCTGCACGGG CTTCTTGGCC TTGTAGGTGG
            541 TCTTGACCTC AGCGTCGTAG TGGCCGCCGT CCTTCAGCTT CAGCCTCTGC TTGATCTCGC
            601 CCTTCAGGGC GCCGTCCTCG GGGTACATCC GCTCAGTGGA GGCCTCCCAG CCCATGGTAC
            661 GACACTGCAT TACGGGGCCG TCGGANGGGA AGTTGGTGCN NNNCAGCTTC ACCTTGTAGA
            721 TGAACTCGCC GTCCTGCANG GAGGAGTCCT GGGTCACGGT CACCACGCCG CCNTCCTCGA
            781 AGTTCATNAN NNNCTCCCNA TTGAAGCCCT CGGGGANNGN NAGCTTCNAG TAGTCNNGGA
            841 NGTNNGCGGG NNGCTTCACG TNNNCCNNNN ANNNNNACAT GANNNNNNGG GGNNAGGNNG
            901 NNCCANGNNN NNGNNNGNGG NCNNNCNTGG NNNNNCTTCN NCNNGNCNGT CNGGGTGNCN
          961 NNNNNNNGGC NGNCCNNNNN NNNNN
          //
          Primer: PGKpro_F (Pr0249)
          Region: insert mid( HygR 5')
          Sequence file: RDB13936_A6B8l.seq check
          >D03070A20_A6B8_1_PGK-proF_B08_05.ab1
              1 NNNNNNNNNN NNNNNNNNNN TCTCCNGGCC TTTCGACCTG CATCCNTCTA GATCTCGAGC
             61 AGCTGAAGCT TACCGCTAGC ATGGATAGAT CCGGAAAGCC TGAACTCACC GCGACGTCTG
            121 TCGAGAAGTT TCTGATCGAA AAGTTCGACA GCGTCTCCGA CCTGATGCAG CTCTCGGAGG
            181 GCGAAGAATC TCGTGCTTTC AGCTTCGATG TAGGAGGGCG TGGATATGTC CTGCGGGTAA
            241 ATAGCTGCGC CGATGGTTTC TACAAAGATC GTTATGTTTA TCGGCACTTT GCATCGGCCG
            301 CGCTCCCGAT TCCGGAAGTG CTTGACATTG GGGAATTCAG CGAGAGCCTG ACCTATTGCA
            361 TCTCCCGCCG TGCACAGGGT GTCACGTTGC AAGACCTGCC TGAAACCGAA CTGCCCGCTG
            421 TTCTGCAGCC GGTCGCGGAG GCCATGGATG CGATCGCTGC GGCCGATCTT AGCCAGACGA
            481 GCGGGTTCGG CCCATTCGGA CCGCAAGGAA TCGGTCAATA CACTACATGG CGTGATTTCA
            541 TATGCGCGAT TGCTGATCCC CATGTGTATC ACTGGCAAAC TGTGATGGAC GACACCGTCA
            601 GTGCGTCCGT CGCGCAGGCT CTCGATGAGC TGATGCTTTG GGCCGAGGAC TGCCCCGAAG
            661 TCCGGCACCT CGTGCACGCG GATTTCGGCT CCAACAATGT CCTGACGGAC AATGGCCGCA
            721 TAACAGCGGT CATTGACTGG AGCGAGGCGA TGTTCGGGGN NTCCCAATAC GANGTCGCCA
            781 ACATCTTCTT CTGGANGCCG TGGTTGGCTT GTATGGAGCA GCAGACGCGC TACTTCNAGC
            841 GGANGCATCC NGANCTTGCN NNTCGCCGCG GCTCCNGGCG TATATGCTNC GCATTNGTCT
            901 TGANNNACTC TATCNNANCT TGNTNGANNN NNNTTTCGAT GANGCNNCNN GGGNNNNNGN
          961 NCNANGCNNC NNNNNNNNNC NNNNNNNNAN NNNGNNNNNN NNGGNNNNNN NNNN
          //

          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 Tungadi, E.A., Human microcephaly ASPM protein is a spindle pole-focusing factor that functions redundantly with CDK5RAP2. J. Cell Sci. 130 (21): 3676-3684 (2017). PMID 28883092.
          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

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