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

pMK290 (mAID-mClover-Hygro)

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

Catalog number RDB13933
Resource name pMK290 (mAID-mClover-Hygro)
Alternative name mAID-mClover-Hygro, 1025
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.
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.)
          RDB13933 pMK290 (mAID-mClover-Hygro) 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 RDB13933_A6B8p1_v2.pdf check

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

          Primer: M13_-40
          Region: T7 pro,Linker,insert 5'(mAID,mClover)
          Sequence file: RDB13933_A6B8e.seq check
          >D03070A17_A6B8_1_M13(-40)_B05_05.ab1
              1 CCGCCGCTAA 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 GCAGCTCGGC CGACCACTAC CAGCAGAACA
            841 CCCCCATCGG CGACGGCCCC GTGCTGCTGC CCGACAACCA CTACCTGAGC CATCAGTCCA
            901 AGCTGAGCAA AGACCCCAAC GAGAAGCGCG AATCACATGG GTCCCTGCTG GGAGTTCGTG
            961 ACCGCCCGCC CGGGATTACC ACTTGGGCAA TGGGAACGAG CTTGTACCAG GTTAATCCTA
           1021 GATAAACTGA TCATAAATCC AGCGCATAAC CAACAATTTG GTAGAAGGTT TTACTTGGCT
           1081 TAAAAAAACC CTCCCACAAC CTTCCACCCT GAACCTGAAA CACATAAAAA TGAATTGCCA
          1141 ACTGTGGTTG TTAAACCTTG GATTATATTG GCCAGCCTTT AATATATGCG AGTA
          //
          Primer: PGKpro_F
          Region: mouse PGK pro,HygR
          Sequence file: RDB13933_A6B8f.seq check
          >13933_13933_A6Db_PGKpro_F_H03_24.ab1
              1 AGGGCCGTAT CTCTCTTCTC ATCTCCGGGC CTTTCGACCT GCATCCATCT AGATCTCGAG
             61 CAGCTGAAGC TTACCGCTAG CATGGATAGA TCCGGAAAGC CTGAACTCAC CGCGACGTCT
            121 GTCGAGAAGT TTCTGATCGA AAAGTTCGAC AGCGTCTCCG ACCTGATGCA GCTCTCGGAG
            181 GGCGAAGAAT CTCGTGCTTT CAGCTTCGAT GTAGGAGGGC GTGGATATGT CCTGCGGGTA
            241 AATAGCTGCG CCGATGGTTT CTACAAAGAT CGTTATGTTT ATCGGCACTT TGCATCGGCC
            301 GCGCTCCCGA TTCCGGAAGT GCTTGACATT GGGGAATTCA GCGAGAGCCT GACCTATTGC
            361 ATCTCCCGCC GTGCACAGGG TGTCACGTTG CAAGACCTGC CTGAAACCGA ACTGCCCGCT
            421 GTTCTGCAGC CGGTCGCGGA GGCCATGGAT GCGATCGCTG CGGCCGATCT TAGCCAGACG
            481 AGCGGGTTCG GCCCATTCGG ACCGCAAGGA ATCGGTCAAT ACACTACATG GCGTGATTTC
            541 ATATGCGCGA TTGCTGATCC CCATGTGTAT CACTGGCAAA CTGTGATGGA CGACACCGTC
            601 AGTGCGTCCG TCGCGCAGGC TCTCGATGAG CTGATGCTTT GGGCCGAGGA CTGCCCCGAA
            661 GTCCGGCACC TCGTGCACGC GGATTTCGGC TCCAACAATG TCCTGACGGA CAATGGCCGC
            721 ATAACAGCGG TCATTGACTG GAGCGAGGCG ATGTTCGGGG ATTCCAATAC GAGGTCGCCA
            781 ACATCTTCTT CTGGAGGCCG TGGTTGGCTT GTATGGAGCA GCAGACGCGC TACTTCGAGC
            841 GGAGGCATCC GGAGCTTGCA GATCGCCGCG GCTCCGGGCG TATATGCTCC GCATTGGTCT
            901 TGACCAACTC TATCAGAGCT TGGGTTTGAC GGCAATTTCG ATGATGCAGC TTTGGGCGCA
            961 GGGTCGATGC GACGCAATCG TCCCGATCCG GAGCCGGACT GTCGGGCGTA CACAAATCGC
           1021 CCGCAGAAGC GGCGTCGGTC TGACCGGATG GCCTGTGTAG AGTTACTTCG CGAATAGGTG
          1081 CCAAAACCGG AACGG
          //
          Primer: M13R
          Region: T3 pro,loxP,mouse PGK ter.,HygR
          Sequence file: RDB13933_A6B8g.seq check
          >D03070A17_A6B8_1_M13R_B04_04.ab1
              1 CAGGCCGGCA TTACCCTCAC TAAAGGGAAC AAAAGCTGGG TACCGGATCC 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 CTATTCCTTT GCCCTCGGAC GAGTGCTGGG GCGTCGGTTT CCACTATCGG
            721 CGAGTACTTC TACACAGCCA TCGGTCCAGA CGGCCGCGCT TCTGCGGGCG ATTTGTGTAC
            781 GCCCGACAGT CCCGGCTCCG GATCGGACGA TTGCGTCGCA TCGACCCTGC GCCCAAGCTG
            841 CATCATCGAA ATTGCCGTCA ACCAAGCTCT GATAGAGTTG GTCAAGACCA ATGCGGAGCA
            901 TATACGCCCG GAGCCGCGGC GATCCTGCAA GCTCCGGATG CCTCCGCTCG AAGTAGCGCG
            961 TCTGCTGCTC CATACAAGCC AACCCACGGC CTCCAGAAGA AGATGTTGGG CGGACCTCGT
           1021 ATTGGGGAAA TCCCCCGAAC ATCGCCTCGC CTCCAGTCAA ATGACCCGCT GGTATGGCCG
           1081 GCCATGGTCG TCAGGGACAT TGGTGGAGTG AATCCGCGGT GCACGAAGGG TGGCCGGAAC
          1141 CTTCCGGGGG GC
          //
          Primer: PGK_N_rev
          Region: mouse PGK pro,loxP,SV40pA,insert 3'(mClover)
          Sequence file: RDB13933_A6B8h.seq check
          >D03070A17_A6B8_1_PGK-N-rev_E06_15.ab1
              1 CATGGGGAAC GCCTCCCTAC CGGTAGAATT TCGAGGTCGA GATCCTAAGC TTGGCTGGAC
             61 GTAAACTCCT CTTCAGACCT AATAACTTCG TATAGCATAC ATTATACGAA GTTATATTAA
            121 GGGTTATTGA ATATGATCGA CGCGTTAAGA TACATTGATG AGTTTGGACA AACCACAACT
            181 AGAATGCAGT GAAAAAAATG CTTTATTTGT GAAATTTGTG ATGCTATTGC TTTATTTGTA
            241 ACCATTATAA GCTGCAATAA ACAAGTTAAC AACAACAATT GCATTCATTT TATGTTTCAG
            301 GTTCAGGGGG AGGTGTGGGA GGTTTTTTAA AGCAAGTAAA ACCTCTACAA ATGTGGTATG
            361 GCTGATTATG ATCAGTTATC TAGATTACTT GTACAGCTCG TCCATGCCAT GTGTAATCCC
            421 GGCGGCGGTC ACGAACTCCA GCAGGACCAT GTGATCGCGC TTCTCGTTGG GGTCTTTGCT
            481 CAGCTTGGAC TGATGGCTCA GGTAGTGGTT GTCGGGCAGC AGCACGGGGC CGTCGCCGAT
            541 GGGGGTGTTC TGCTGGTAGT GGTCGGCGAG CTGCACGCTG CCGTCCTCAA CGTTGTGGCG
            601 GATCTTGAAG TTAGCCTTGA TGCCGTTCTT CTGCTTGTCG GCCGTGATAT AGACGTTGTG
            661 GCTGTTGAAG TTGTACTCCA GCTTGTGCCC CAGGATGTTG CCGTCCTCCT TGAAAGTCGA
          721 TGCCCTTCAG CTCGATGCGG TTCACCAGGG GTGTCGCCCT
          //

          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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