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

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.
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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          RDB13935 pMK292 (mAID-mCherry2-NeoR) 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 pMK292 (mAID-mCherry2-NeoR) was provided by the RIKEN BRC through the National BioResource Project of the MEXT, Japan (cat. RDB13935).

          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 RDB13935_A6B8p1-4.pdf check

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

          Primer: M13_-40 (Pr0410)
          Region: insert 5'(Linker,mAID,mCherry2 5')
          Sequence file: RDB13935_A6B8i.seq check
          >D03070A19_A6B8_1_M13(-40)_D05_11.ab1
              1 NNNNNNNNNN NNNNNNNNCN NNNNNNGANT NNNNNTAGGG CGAANTGGAG CTCCCCGGGN
             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 GACNNNNNAC TCCTCCCTGC AGGACGGCGA GTTCATCTAC
            661 AAGGTGAAGC TGCGCGGCAC CNACTTCCCC TCCGACGGCC CCGTAATGCA GTGTCGTACC
            721 ATGGGCTGGG AGGCCTCCAC TGNNNNGATG TNNNCCCGAG GACGGCGCCC TGAANGGCGA
            781 GATCNAGCAG ANGCTGAANG CTGANNGACG GCGGCCACTA CNACGCTGAN GTCAAGACCA
            841 CCTANNNNGC CNAGAAGCCC GTGCAGCTGN CCNGCGCNNN NNACGTCNAN NNCNNGTTNG
            901 GANATCCTTN NNNNNNNCNN GGACTANNNC NTCGNGGAAN NANNNNNNAA NNNNNNNNNG
            961 GGNCGNCNNC TNNNNCNGNN GGCNNNGNNN NNNCNGTNCA NNNNNTCTNN NANNANCTGA
           1021 NNNNNNANNC NNNNNTNNCA CATTTNNNNN NANGGNTTTT ANNNNNNNTT TNNNNNNNNN
          1081 N
          //
          Primer: M13R (Pr0160)
          Region: insert 3'(NeoR_KanR 3')
          Sequence file: RDB13935_A6B8j.seq check
          >D03070A19_A6B8_1_M13R_D04_10.ab1
              1 NNNNNNNNNN NNNNNNNNNN ANGGGNNNNN AAGCTGGGTA NCGGATCCNA 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 CGTCNAGCAC AGCTGCNCAN NGAACGCCCG
            901 NCGNGGCCAG CCACGATAGC CGCGCNGCCN CGTCTNGCAG TTCANTCNNN GCANCNGNNN
          961 GGTCNGTCTN GANNAAAANA NNCNGGCGCC CNNNCNNNNN NNNNCNNNNN NNNNN
          //
          Primer: SV40polyA_R2 (Pr0623)
          Region: insert mid(SV40 ter,mCherry2 3')
          Sequence file: RDB13935_A6B8k.seq check
          >13935_13935_A9KsF_SV40polyA_R2_F09_18_ABI24.ab1
              1 NNNNNNNNGN ANNANNNNGT TAACAACAAC AATTGCATTC ATTTTATGTT TCNGGTTCAG
             61 GGGGAGGTGT GGGAGGTTTT TTAAAGCAAG TAAAACCTCT ACAAATGTGG TATGGCTGAT
            121 TATGATCAGT TATCTAGATT ACTTGTACAG CTCGTCCATG CCGCCGGTGG AGTGGCGGCC
            181 CTCGGCGCGT TCGTACTGTT CCACGATGGT GTAGTCCTCG TTGTGGGAAA GGATGTCCAA
            241 CTTGATGTCG ACGTTGTAGG CGCCGGGCAG CTGCACGGGC TTCTTGGCCT TGTAGGTGGT
            301 CTTGACCTCA GCGTCGTAGT GGCCGCCGTC CTTCAGCTTC AGCCTCTGCT TGATCTCGCC
            361 CTTCAGGGCG CCGTCCTCGG GGTACATCCG CTCAGTGGAG GCCTCCCAGC CCATGGTACG
            421 ACACTGCATT ACGGGGCCGT CGGAGGGGAA GTTGGTGCCG CGCAGCTTCA CCTTGTAGAT
            481 GAACTCGCCG TCCTGCAGGG AGGAGTCCTG GGTCACGGTC ACCACGCCGC CGTCCTCGAA
            541 GTTCATCACG CGCTCCCAAT TGAAGCCCTC GGGGAAGGAC AGCTTCAAGT AGTCGGGGAT
            601 GTCGGCGGGG TGCTTCACGT AGGCCTTGGA GCCGTACATG AACTGAGGGG ACAGGATGTC
            661 CCAGGCGAAG GGCAGGGGGC CACCCTTGGT CACCTTCAGC TTGNCGGTCT GGGTGCCCTC
            721 GTANGGGCGG NCCTCGCCCT CGCCCTCGAT CTCGAACTCG TGGCCGTTCA CGGAGCCCTC
            781 CATGTGCANC TTGAAGCGCA TGAACTCCTT GATGATGGCC ATGTTATCCT CCTCGCCCTT
            841 GCTCACCATG CTAGCTTTAT ACATCCTCAA NTCGATTTTC CTCNAGTACN GNGCNNCGNN
            901 NATTGATNNC TTCACNAACG CCGCCGCNNN CNGGNCACCG CTNGANTTTT GGNNNNAANN
          961 NNTCNCGTTC NNNCGGTATG ATNNNNNNNN NN
          //
          Primer: SV40pro_F_V2 (Pr0154)
          Region: insert mid(NeoR_KanR 5 ')
          Sequence file: RDB13935_A6B8l.seq check
          >D03070A19_A6B8_1_SV40pro_F_V2_F10_16.ab1
              1 NNNNNNNNNN NNNNNNNNNN NNNGCTTTTG CAAGNNCGAT CNAGAGANNG GATGAGGNTC
             61 GTTTCGCATG ATTGAACNNG 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 TGCTGANNAG CTTGGCGGCG AATGGGCTGA
            781 CCGCTTCCTC GTGCTTTACG GTATCGCCGC TNCCGATTCG CAGCGCATCG CCTTCTATCG
            841 CCTTCTTGAC GANTTCTTCT GAGCGGGNNT CTGGGGTTCG AANTGACCGA NCAAGCGNNG
            901 CCCNANCTGN CNTCNCGANN NTNCNATTCC NCCGCCGCCT TCTATGAANN NNNGGGGCTT
          961 NNNNNNNNNT TTNCNNGNNN NNNNGNTNNN NGNNNNNNNN NN
          //

          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 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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