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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 set forth by the DEPOSITOR 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.
          Ordering Please visit Information of Request for Distribution.[link] 
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          Material Transfer Agreement (MTA for use for not-for-profit academic purpose) [Word]
          This RESOURCE contains a fluorescent protein owned or licensed by Global Life Sciences Solutions Operations UK Ltd. (Cytiva). Please refer to Schedule B and use Schedule A (download the Schedule A and Schedule B [link]).
          提供案内 (日本国内) [open/close]

          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。
          提供依頼 手続きの詳細は、「提供申込みについて[link]」をご覧ください。
          提供依頼書 [Word]
          提供同意書 (MTA、非営利機関による非営利学術研究用)[Word]
          GFPリソースを含みます。Schedule Bをご一読の上Schedule A (英語版のみ) の提出をお願いします (Schedule AとSchedule Bを入手[link])。

          Catalog # Resource name Shipping form Fee (non-profit org.)
          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.

          References and tips

          Electronic file

          Original reference

          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.

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

          Featured content

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

          Sequence information

          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_v2.pdf check

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

          Primer: M13_-40
          Region: insert5'(T7pro,Linker,mAID,mClover(A206K))
          Sequence file: RDB13932_A6B8e.seq check
          >D03070A16_A6B8_1_M13(-40)_A05_02.ab1
              1 AAGGCAGCTA CGCGCGTATA CGACTCACTA TAGGGCGAAT TGGAGCTCCC CGGGATCCGG
             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 GGCATCAAGG CTAACTTCAA
            781 GATCCGCCAC AACGTTGAGG ACGGCAGCGT GCAGCTCGCC GACCACTACC AGCAGAACAC
            841 CCCCATCGGC GACGGCCCCG TGCTGCTGCC CGACAACCAC TACCTGAGCC ATCAGTCCAA
            901 GCTGAGCAAA GACCCCAACG AGAAGCGCGA TCACATGGTC CTGCTGGAGT TCGTGACCGC
            961 CGCCCGGGAT TACACATGGC ATGGAACGAG CTGTACAAGT AATCTAGATA ACTGATCATA
           1021 TCAGCCCATA CCACATTTGT AGAGGTTTTA CTGCTTAAAA CCTCCCACAC TCCCCTTGAA
           1081 CTTGAAACAT AAAATGAATG CCATGATGGT GCTACCTGTC TATGCGACTT AAAATGGTCA
          1141 TAGCCATAGC CATTCACGAT
          //
          Primer: SV40polyA-R
          Region: insert mid(mClover(A206K))
          Sequence file: RDB13932_A6B8f.seq check
          >D03070A16_A6B8_1_SV40polyA-R_H04_22.ab1
              1 TCCCCCCCCC CGGTATTTAG CAGTAACCTC TACAATGTTG GTATGGGCTG ATTATGATCA
             61 GTTATCTAGA TTACTTGTAC AGCTCGTCCA TGCCATGTGT AATCCCGGCG GCGGTCACGA
            121 ACTCCAGCAG GACCATGTGA TCGCGCTTCT CGTTGGGGTC TTTGCTCAGC TTGGACTGAT
            181 GGCTCAGGTA GTGGTTGTCG GGCAGCAGCA CGGGGCCGTC GCCGATGGGG GTGTTCTGCT
            241 GGTAGTGGTC GGCGAGCTGC ACGCTGCCGT CCTCAACGTT GTGGCGGATC TTGAAGTTAG
            301 CCTTGATGCC GTTCTTCTGC TTGTCGGCCG TGATATAGAC GTTGTGGCTG TTGAAGTTGT
            361 ACTCCAGCTT GTGCCCCAGG ATGTTGCCGT CCTCCTTGAA GTCGATGCCC TTCAGCTCGA
            421 TGCGGTTCAC CAGGGTGTCG CCCTCGAACT TCACCTCGGC GCGGGTCTTG TAGGTACCGT
            481 CGTCCTTGAA AGAGATGGTG CGCTCCTGGA CGTAGCCTTC GGGCATGGCG GACTTGAAGA
            541 AGTCGTGCTG CTTCATGTGG TCGGGGTACC GGCTGAAGCA GGCCACGCCG TAGCCGAAGG
            601 TGGTCACGAG GTGGGCCAGG GCACGGGCAG CTTGCCGGTG GTGCAGATGA ACTTCAGGGT
          661 CAGCTTGCCG TTG
          //
          Primer: SV40pro_F_V2
          Region: insert mid(NeoR_KanR)
          Sequence file: RDB13932_A6B8g.seq check
          >D03070A16_A6B8_1_SV40pro_F_V2_E10_13.ab1
              1 GGGGCAGCTT TTGGAGGCCT AGGCTTTTGC AAGATCGATC AAGAGACAGG ATGAGGATCG
             61 TTTCGCATGA TTGAACAAGA 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 GGACCGCTAT
            721 CAGGACATAG CGTTGGCTAC CCGTGATATT GCTGAAGAGC TTGGCGGCGA ATGGGCTGAC
            781 CGCTTCCTCG TGCTTTACGG TATCGCCGCT CCCGATTCGC AGCGCATCGC CTTCTATCGC
            841 CTTCTTGACG AGTTCTTCTG AGCGGGACTC TGGGGTTCGA AATGACCGAC CAAGCGACGC
            901 CCAACCTGCC ATCACGAGAT TTCGATTCCA CCGCCGCCTT CTATGAAAGG TTGGGCTTCG
            961 GAATCGTTTT CCGGGACGCC CGGCTGGATG ATCCCTCCAG CGCGGGGATC TCATGCTGGG
           1021 AGTTCTTTCG CCCACCCTAG GGGAGGCTAA CTGAAACACG GAAGAGACAA TACCGGAAAG
          1081 ACCGCGCTAT GACGGCCAAT AAAAGGACAG GAAATAAAAC CGCCCACCG
          //
          Primer: M13R
          Region: insert3'(TKp(A),NeoR_KanR)
          Sequence file: RDB13932_A6B8h.seq check
          >D03070A16_A6B8_1_M13R_A04_01.ab1
              1 CTGGGCGCAA TTAACCTCAC TAAAGGGAAC AAAAGCTGGG TACCGGATCC AACGACCCAA
             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 GGTGAGATGA CAGGAGATCC TGCCCCGGCA CTTCGCCCAA
            841 TAGCAGCCAG TCCCTTCCCG CTTCAGTGAC AACGTCGAGC ACAGCTGCGC AAGGAACGCC
            901 CCGTCGTGGC CCAGCCACGA TAGCCGCGCT GCCTCGTCTT GCAGTTCATT CAAGGCACCG
            961 GGACAGGGTC GGGTCTTTGA CAAAAAGAAT CCGGGGCGGC CCCTGCGCTT GACAGGCCGG
           1021 GAACCACGGG CGGGCAATCA GAAGCAGCCC GAATTGTCTG TTGGTGGCCC AGTCAATAGC
           1081 CGGAAATAGG CTCTCCACCC CAAGCCGGGC CGGGAGGAAC TGCGTGCCAA TTCCAATCCT
          1141 TTGGCTCATT CAATGCCGGA AAACGGAATT CCCTTCACAT CACCTCTG
          //

          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.
          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.

          2022.05.17

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