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

pMK289 (mAID-mClover-NeoR)

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

Catalog number RDB13932
Resource name pMK289 (mAID-mClover-NeoR)
Alternative name mAID-mClover-NeoR, 1002
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. 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 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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          RDB13932 pMK289 (mAID-mClover-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 pMK289 (mAID-mClover-NeoR) was provided by the RIKEN BRC through the National BioResource Project of the MEXT, Japan (cat. RDB13932).

          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 RDB13932_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, mClover 5')
          Sequence file: RDB13932_A6B8i.seq check
          >D03070A16_A6B8_1_M13(-40)_A05_02.ab1
              1 NNNNNNNNNN NNCNCNNANN NGNNTNNCNN TAGGGCGAAT 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 GCAGAAGAAC GGCATCAANG CTAACTTCAA
            781 GATCCGCCAC AACGTTGANG ANGGNAGCGN GCANCTCGCC GACCACTACC AGCAGANCAC
            841 CCCCATCGGC GACGGNCCCG TGCTGCTGNC CGANNACCAC TACCTNANCC ATCAGNCCAA
            901 GCTGANCAAA GANCCNNACG AGAANCGCGA TNNCNTGGNC CNGCTNNNNN NNNNGANNNN
          961 NNNCNNGNNN NNNNNNTGGN NNNNNNNNAN NNNNNNNNN
          //
          Primer: M13R (Pr0160)
          Region: insert 3'(NeoR_KanR 3')
          Sequence file: RDB13932_A6B8j.seq check
          >D03070A16_A6B8_1_M13R_A04_01.ab1
              1 NNNNNNNNNN NNNNNNNCNN NNNNGGGAAN NNAAGCTGGG TACCGGATCC NACGACCCAA
             61 CACCGTGCGT TTTATTCTGT CTTTTTATTG CCGTCATAGC GCGGGTTCCT TCCGGTATTG
            121 TCTCCTTCCG TGTTTCAGTT AGCCTCCCCC TAGGGTGGGC GAAGAACTCC AGCATGAGAT
            181 CCCCGCGCTG GAGGATCATC CAGCCGGCGT CCCGGAAAAC GATTCCGAAG CCCAACCTTT
            241 CATAGAAGGC GGCGGTGGAA TCGAAATCTC GTGATGGCAG GTTGGGCGTC GCTTGGTCGG
            301 TCATTTCGAA CCCCAGAGTC CCGCTCAGAA GAACTCGTCA AGAAGGCGAT AGAAGGCGAT
            361 GCGCTGCGAA TCGGGAGCGG CGATACCGTA AAGCACGAGG AAGCGGTCAG CCCATTCGCC
            421 GCCAAGCTCT TCAGCAATAT CACGGGTAGC CAACGCTATG TCCTGATAGC GGTCCGCCAC
            481 ACCCAGCCGG CCACAGTCGA TGAATCCAGA AAAGCGGCCA TTTTCCACCA TGATATTCGG
            541 CAAGCAGGCA TCGCCATGGG TCACGACGAG ATCCTCGCCG TCGGGCATGC TCGCCTTGAG
            601 CCTGGCGAAC AGTTCGGCTG GCGCGAGCCC CTGATGCTCT TCGTCCAGAT CATCCTGATC
            661 GACAAGACCG GCTTCCATCC GAGTACGTGC TCGCTCGATG CGATGTTTCG CTTGGTGGTC
            721 GAATGGGCAG GTAGCCGGAT CAAGCGTATG CAGCCGCCGC ATTGCATCAG CCATGATGGA
            781 TACTTTCTCG GCAGGAGCAA NGTGAGATGA CAGGAGATCC TGCCCCGGCA CTTCGCCCAA
            841 TAGCAGCNAG TNCCTTCCCG CTTCANTGAC AACGTCGAGC ACAGCTGCNC NNNGAACGCC
            901 CCGTCNNNGN CNAGCNNCNA NAGCCGCGCN GCCNCGTCTT GCAGTTNATT CNNGGNNNCN
            961 GNACANGGTC NGNNNNTNGN NAAAAANNAN NNNGGGNNNC CCCNNCNNNN NANNNNCNNG
          1021 NNANNANNNN CNGNCNNNNA NNNNNNNNNN
          //
          Primer: SV40polyA-R (Pr0109)
          Region: insert mid(SV40 pA,mClover 3')
          Sequence file: RDB13932_A6B8k.seq check
          >D03070A16_A6B8_1_SV40polyA-R_H04_22.ab1
              1 NNNNNNNNNN NNNNNNNNNN NNNNNNCCTC NNNNNNGNNG GNNNNNGCTG ATTANGNTNN
             61 NTTATCTAGA TTACTTGNNN NNNTCGTCCA TGCCATGTGT AATCCCGGCG GCGGTCACNA
            121 ACTCCANCNN GACCATGTGA TCGCGCTTCT CGNNNNNGTC TTTGCTCANC TTGGACTGAT
            181 GGCTCANGTA GNGNNNGTCG GGCAGCANCA CGGGGCCGTC GCCGATGGGG GTGTTCTGCT
            241 GGTAGTGGTC GGCNAGCTGC ACNCTGCCGT CCTCAACGTT GTGGCGGATC TTGNNNNTNN
            301 CCTTGATGCC GTTCTTCTGC TTGTNNNNNN TGATATAGAC NTTGNGNCTG TNNANNNTGT
            361 ACTCCANCTT GTGCCCCANG NNNNNNCCGT CCTCCNNNNN NTCNATGCCC TTCAGCTCGA
            421 TGCGGTNNNN NNNNGTGTNN NCNTCNAACT NNNNNNNNGC NCGNNNNNNG TANGTNCCGT
          481 CNTCNTNNAN AGANANNNNN NNNNN
          //
          Primer: SV40pro_F_V2 (Pr0154)
          Region: insert mid(NeoR_KanR5 ')
          Sequence file: RDB13932_A6B8l.seq check
          >D03070A16_A6B8_1_SV40pro_F_V2_E10_13.ab1
              1 NNNNNNNNNN NNNNNNNNNN NNGCTTTNGN NNNNNCGATC NAGAGNNAGG NNGAGGNNCG
             61 TTTCGCATGA TTGAACNAGA TGGATTGCAC GCAGGTTCTC CGGCCGCTTG GGTGGAGAGG
            121 CTATTCGGCT ATGACTGGGC ACAACAGACA ATCGGCTGCT CTGATGCCGC CGTGTTCCGG
            181 CTGTCAGCGC AGGGGCGCCC GGTTCTTTTT GTCAAGACCG ACCTGTCCGG TGCCCTGAAT
            241 GAACTGCAAG ACGAGGCAGC GCGGCTATCG TGGCTGGCCA CGACGGGCGT TCCTTGCGCA
            301 GCTGTGCTCG ACGTTGTCAC TGAAGCGGGA AGGGACTGGC TGCTATTGGG CGAAGTGCCG
            361 GGGCAGGATC TCCTGTCATC TCACCTTGCT CCTGCCGAGA AAGTATCCAT CATGGCTGAT
            421 GCAATGCGGC GGCTGCATAC GCTTGATCCG GCTACCTGCC CATTCGACCA CCAAGCGAAA
            481 CATCGCATCG AGCGAGCACG TACTCGGATG GAAGCCGGTC TTGTCGATCA GGATGATCTG
            541 GACGAAGAGC ATCAGGGGCT CGCGCCAGCC GAACTGTTCG CCAGGCTCAA GGCGAGCATG
            601 CCCGACGGCG AGGATCTCGT CGTGACCCAT GGCGATGCCT GCTTGCCGAA TATCATGGTG
            661 GAAAATGGCC GCTTTTCTGG ATTCATCGAC TGTGGCCGGC TGGGTGTGGC GGANCGCTAT
            721 CANGACATAG CGTTGGCTAC CCGTGATATT GCTGAAGAGC TTGGCGGCGA ATGGGCTGAC
            781 CGCTTCCTCG TGCTTTACGG TATCGCCGCT CCCGATTCGC ANCGCATCGC CTTCTATCGN
            841 CTTCTTGACG ANTTCTTCTG ANCGGGANTC TGGGGNTCGA AATGNNCGNN CNNNCGACGC
            901 CCAACCNGNC NTCACGANAN TNCGANNCCN CCGCCGNNTT CTATGAAAGN NNNGNNTNNN
          961 NAATCGTTTT NNGGGANNNN NNNNNN
          //

          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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