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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 |
Other clones in our bank

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 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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          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.

          How to cite this biological resource

          Materials & Methods section:

          The pMK286 (mAID-NeoR) was provided by the RIKEN BRC through the National BioResource Project of the MEXT, Japan (cat. RDB13929).

          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 RDB13929_A6B8p1-4.pdf check

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

          Primer: pUC-M13_-100 (Pr0310)
          Region: insert5'(T7 pro,Linker,mAID,SV40 pA,f1 ori)
          Sequence file: RDB13929_A6B8g.seq check
          >13929_13929_B3BoF_pUC-M13_-100_F01_06_ABI08.ab1
              1 NNNNNNNNNN NNNNNNNNNN NNGCNNGNNN TTTCCNNGTC NCGNCGTTGT AAAACGACGG
             61 CCAGTGAGCG CGCGTAATAC GACTCACTAT AGGGCGAATT GGAGCTCCCC GGGATCCGGT
            121 GCAGGCGCCA AGGAGAAGAG TGCTTGTCCT AAAGATCCAG CCAAACCTCC GGCCAAGGCA
            181 CAAGTTGTGG GATGGCCACC GGTGAGATCA TACCGGAAGA ACGTGATGGT TTCCTGCCAA
            241 AAATCAAGCG GTGGCCCGGA GGCGGCGGCG TTCGTGAAGG TATCAATGGA CGGAGCACCG
            301 TACTTGAGGA AAATCGATTT GAGGATGTAT AAATAATCTA GATAACTGAT CATAATCAGC
            361 CATACCACAT TTGTAGAGGT TTTACTTGCT TTAAAAAACC TCCCACACCT CCCCCTGAAC
            421 CTGAAACATA AAATGAATGC AATTGTTGTT GTTAACTTGT TTATTGCAGC TTATAATGGT
            481 TACAAATAAA GCAATAGCAT CACAAATTTC ACAAATAAAG CATTTTTTTC ACTGCATTCT
            541 AGTTGTGGTT TGTCCAAACT CATCAATGTA TCTTAACGCG TAAATTGTAA GCGTTAATAT
            601 TTTGTNNNNN NTCGCGTTAA ATTTTTGTTA AATCANCTCA TTTTTTAACC AATAGGCCGA
            661 AAATCGGCAA AATCCCTTAT AAATCAAAAA GAATAGACCG AGATAGGGTT GAGTGTTGTT
            721 CCNGTTTGGA ACAAGAGTCC ACTATTANNN NNNCGTGGAC TCNNACGTNN AAGGGCGAAA
            781 AAANCNTCTA TCAGGNNGAT GGCCCNNTAC GTGNANCCAT CNCCCNTANT CAAGTTTTTT
            841 NGGGGGNNNA NGTGNCNTNN NNNNNNNAAA TNCGGAACCC NNAAAGGGNN NNCCCCNNNA
            901 NTTTNNAGCT TGNNNNGGNN AANNCNNCNN ANCNTGGNNN ANNAAAGNNA NNGNNNNNNN
          961 NNN
          //
          Primer: AFR (Pr0324)
          Region: insert3'(T3 pro,TK pA,NeoR_KanR)
          Sequence file: RDB13929_A6B8h.seq check
          >13929_13929_B3BoF_AFR_G01_07_ABI08.ab1
              1 NNNNNNNNNN NNNNNNNNNN NGCNNNGCGC GCANTTAACC CTCACTAAAG GGAACAAAAG
             61 CTGGGTACCG GATCCAACGA CCCAACACCG TGCGTTTTAT TCTGTCTTTT TATTGCCGTC
            121 ATAGCGCGGG TTCCTTCCGG TATTGTCTCC TTCCGTGTTT CAGTTAGCCT CCCCCTAGGG
            181 TGGGCGAAGA ACTCCAGCAT GAGATCCCCG CGCTGGAGGA TCATCCAGCC GGCGTCCCGG
            241 AAAACGATTC CGAAGCCCAA CCTTTCATAG AAGGCGGCGG TGGAATCGAA ATCTCGTGAT
            301 GGCAGGTTGG GCGTCGCTTG GTCGGTCATT TCGAACCCCA GAGTCCCGCT CAGAAGAACT
            361 CGTCAAGAAG GCGATAGAAG GCGATGCGCT GCGAATCGGG AGCGGCGATA CCGTAAAGCA
            421 CGAGGAAGCG GTCAGCCCAT TCGCCGCCAA GCTCTTCAGC AATATCACGG GTAGCCAACG
            481 CTATGTCCTG ATAGCGGTCC GCCACACCCA GCCGGCCACA GTCGATGAAT CCAGAAAAGC
            541 GGCCATTTTC CACCATGATA TTCGGCAAGC AGGCATCGCC ATGGGTCACG ACGAGATCCT
            601 CGCCGTCGGG CATGCTCGCN NNNNNCCTGG CGAACAGTTC GGCTGGCGCG AGCCCCTGAT
            661 GCTCTTCGTC CAGATCATCC TGATCGACAA GACCGGCTTC CATCCGAGTA CGTGCTCGCT
            721 CGATGCGATG TTTCGCTNGG TGGTCGAATG GGCAGGTAGC CGGATCAAGC GTATGCAGCC
            781 GCCGCATTGC ATCAGCCATG ATGGATACTT TCTCGGCAGG ANCANNGTGA GATGACAGGA
            841 GATCCTGCCC CGGNACTTCG CCNNATAGCA GCCAGTCCCT TCCCGCTTCA NNGNCNNCGT
            901 CNNNCACANC TGCNCAAGGA ACGCCNGTCG NNNNNNNGCC NCGATANNCN NCGCNNCNNN
          961 NNCNTNNNNN NTNNNNTNNN GNNNNNNNNN N
          //
          Primer: SV40pro_F_V2 (Pr0154)
          Region: insert mid(Neo_KanR)
          Sequence file: RDB13929_A6B8i.seq check
          >D03070A13_A6B8_1_SV40pro_F_V2_D10_10.ab1
              1 NNNNNNNNNN NNNNNNNNNN NNNGCTTTTG CAAGNNCGAT CAAGANNNAG GNTGAGGNTC
             61 GTTTCGCATG ATTGAACNNN 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 CCTTCTATCN
            841 NCTTCNNGAC GAGTTCTTCT GANCGGGANT CTGGGGGTTC NAAATGNNCG ACCNNGCGNC
            901 GCCCNANCCT GCCNNNNCNN NATTTNNANN NCNNCGCCGC CNNCTATGAA ANNNTNNGGN
            961 CTTCGGANNN NTTTTNNNGG ANNNNNGNCN GGNTGATCCN NCNNNCGNNG GGNNNNNNNN
          1021 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 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.

          2023.10.05

          GNP_filter3_RDBDEP_html_231001.pl