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

pMK286 (mAID-NeoR)

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

Catalog number RDB13929
Resource name pMK286 (mAID-NeoR)
Alternative name mAID-NeoR, 1006
Clone info. A plasmid for construction of C-terminal mAID 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 Sequence analysis is not reliable around MluI site adjacent to SV40 poly(A) site probably because of tangled tertiary structure induced by unexpected palindrome sequences. 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
Depositor|Developer Kanemaki, Masato |
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External Database
Arabidopsis thaliana AtIAA17

          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.
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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。
          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。
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          提供同意書 (MTA、非営利機関による非営利学術研究用)[Word]

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          RDB13929 pMK286 (mAID-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 RDB13929_A6B8p1-3.pdf check

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

          Primer: M13_-40
          Region: insert5'(T7 pro,Linker,mAID,SV40p(A),f1ori)
          Sequence file: RDB13929_A6B8d.seq check
          >D03070A13_A6B8_1_M13(-40)_F03_18.ab1
              1 AGGCGTTACG CGCGTATACG ACTCACTATA GGGCGAATTG GAGCTCCCCG GGATCCGGTG
             61 CAGGCGCCAA GGAGAAGAGT GCTTGTCCTA AAGATCCAGC CAAACCTCCG GCCAAGGCAC
            121 AAGTTGTGGG ATGGCCACCG GTGAGATCAT ACCGGAAGAA CGTGATGGTT TCCTGCCAAA
            181 AATCAAGCGG TGGCCCGGAG GCGGCGGCGT TCGTGAAGGT ATCAATGGAC GGAGCACCGT
            241 ACTTGAGGAA AATCGATTTG AGGATGTATA AATAATCTAG ATAACTGATC ATAATCAGCC
            301 ATACCACATT TGTAGAGGTT TTACTTGCTT TAAAAAACCT CCCACACCTC CCCCTGAACC
            361 TGAAACATAA AATGAATGCA ATTGTTGTTG TTAACTTGTT TATTGCAGCT TATAATGGTT
            421 ACAAATAAAG CAATAGCATC ACAAATTTCA CAAATAAAGC ATTTTTTTCA CTGCATTCTA
            481 GTTGTGGTTT GTCCAAACTC ATCAATGTAT CTTAACGCGT AAATTGTAAG CGTTAATATT
            541 TTGTTAAAAT TCGCGTTAAA TTTTTGTTAA ATCAGCTCAT TTTTTAACCA ATAGGCCGAA
            601 ATCGGCAAAA TCCCTTATAA ATCAAAAGAA TAGACCGAGA TAGGGTTGAG TGTTGTTCCA
            661 GTTTGGAACA AGAGTCCACT ATTAAAGAAC GTGGACTCCA ACGTCAAAGG GCGAAAAACC
            721 GTCTATCAGG GCGATGGCCC CACTACGTGA ACCATCACCC TAATCAAGTT TTTTGGGGTC
            781 GAGGTGCCTT AAAGCACTAA ATCGGAACCC TAAGGGAGCC CCCCATTTAA ACTTGACGGG
            841 AAAATCCCGC GAACGTTGCC AGAAAGGAAA GGAAAAAAGA CAAAAGGACC GGGAGCTAGG
            901 GCACTAGAAA GGCTAACGGG ACCCCCTGCA CTACCACCAC AAAAGCCGCG CTTACTTTCT
            961 CCTGTCATGG ATTCACGGTG CGAAATTAGT CAGTAGAAGA AAACAGAACC CGATTATCAT
          1021 CTCTTAACAG CAAAAACTCA AAAAATTAAG C
          //
          Primer: SV40pro_F_V2
          Region: insert mid(Sv40pro_ori,Neo_KanR)
          Sequence file: RDB13929_A6B8e.seq check
          >D03070A13_A6B8_1_SV40pro_F_V2_D10_10.ab1
              1 GTACCACTTT TTTGGGAGGC TAGGCTTTTG CAAGATCGAT CAAGAGACAG GATGAGGATC
             61 GTTTCGCATG ATTGAACAAG 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 TGCTGAAGAG CTTGGCGGCG AATGGGCTGA
            781 CCGCTTCCTC GTGCTTTACG GTATCGCCGC TCCCGATTCG CAGCGCATCG CCTTCTATCG
            841 CCTTCTTGAC GAGTTCTTCT GAGCGGGACT CTGGGGGTTC GAAATGACCG ACCAAGCGAC
            901 GCCCAAACCT GCCATCACGA GATTTCGATT CCACCGCCGC CTTCTATGAA AAGGTTGGGG
            961 CTTCGGAATC GTTTTTCGGG ACGCCGGGCT GGATGATCCT CCAAGCGCGG GGGATCTCCA
           1021 TGCTGGGAGG TTCTTTCGCC CCACCCCTAG GGGGAGGGCT TACTTGAGAC ACGGGAAGGA
           1081 GACAATACCG GAAGGACCGC GCTATGAACG GCAATTAAAA GGACAGAACT TAAAACGCGC
          1141 CCACCCG
          //
          Primer: M13R
          Region: insert3'(TKp(A),NeoR_KanR)
          Sequence file: RDB13929_A6B8f.seq check
          >D03070A19_A6B8_1_M13R_D04_10.ab1
              1 GCCGCGGCAT TACCTCACTA AAGGGAACAA AAGCTGGGTA CCGGATCCAA 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 CGTCGAGCAC AGCTGCGCAA GGAACGCCCG
            901 TCGTGGCCAG CCACGATAGC CGCGCTGCCT CGTCTTGCAG TTCATTCAGG GCACCGGACA
            961 GGTCGGTCTT GACAAAAAGA ACCGGGCGCC CCTGCGCTGA CAGCCCGGAA CACGCGGCAT
           1021 CAGAGCAGCC CGATTGTCTG TGTGCCCAGT CATAGGCCGG ATAGGCCTCT CCACCCCAAG
           1081 GCGGGCCAGG AGATCTTGCG ATGCCAGTTC CAATTCTTGA TCAGTCATGC CGAAAACCGA
          1141 TTCCCT
          //

          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 Chen, F.X., PAF1 regulation of promoter-proximal pause release via enhancer activation. Science 357 (6357): 1294-1298 (2017). PMID 28860207.
          reference Natsume, T., Acute inactivation of the replicative helicase in human cells triggers MCM8-9-dependent DNA synthesis. Genes Dev. (2017). PMID 28487407.
          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.

          2022.05.17

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