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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.
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 Kanemaki, Masato |
 
Other clones in our bank Arabidopsis thaliana AtIAA17 (KEGG orthology K14484) |

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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RDB13929 pMK286 (mAID-NeoR) DNA solution

cDNA Request [in Japanese] [in English]

References and tips

Electronic file

Featured content

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

References

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.

User reports and related articles (go to bottom)


Sequence information

RIKEN BRC has sequenced portions of this material for quality test. Primers and the results are shown below.

Data are summerized on test sheet below.

Test sheet RDB13929_A6B8p1_v2.pdf

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

Primer: M13_-40 RDB13929_A6B8a_2.seq
>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 RDB13929_A6B8b_2.seq
>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 RDB13929_A6B8c_2.seq
>D03070A13_A6B8_1_M13R_B03_06.ab1
    1 GTCGCGCAGT ACCTCACTAA AGGGAACAAA AGCTGGGTAC CGGATCCAAC GACCCAACAC
   61 CGTGCGTTTT ATTCTGTCTT TTTATTGCCG TCATAGCGCG GGTTCCTTCC GGTATTGTCT
  121 CCTTCCGTGT TTCAGTTAGC CTCCCCCTAG GGTGGGCGAA GAACTCCAGC ATGAGATCCC
  181 CGCGCTGGAG GATCATCCAG CCGGCGTCCC GGAAAACGAT TCCGAAGCCC AACCTTTCAT
  241 AGAAGGCGGC GGTGGAATCG AAATCTCGTG ATGGCAGGTT GGGCGTCGCT TGGTCGGTCA
  301 TTTCGAACCC CAGAGTCCCG CTCAGAAGAA CTCGTCAAGA AGGCGATAGA AGGCGATGCG
  361 CTGCGAATCG GGAGCGGCGA TACCGTAAAG CACGAGGAAG CGGTCAGCCC ATTCGCCGCC
  421 AAGCTCTTCA GCAATATCAC GGGTAGCCAA CGCTATGTCC TGATAGCGGT CCGCCACACC
  481 CAGCCGGCCA CAGTCGATGA ATCCAGAAAA GCGGCCATTT TCCACCATGA TATTCGGCAA
  541 GCAGGCATCG CCATGGGTCA CGACGAGATC CTCGCCGTCG GGCATGCTCG CCTTGAGCCT
  601 GGCGAACAGT TCGGCTGGCG CGAGCCCCTG ATGCTCTTCG TCCAGATCAT CCTGATCGAC
  661 AAGACCGGCT TCCATCCGAG TACGTGCTCG CTCGATGCGA TGTTTCGCTT GGTGGTCGAA
  721 TGGGCAGGTA GCCGGATCAA GCGTATGCAG CCGCCGCATT GCATCAGCCA TGATGGATAC
  781 TTTCTCGGCA GGAGCAAGGT GAGATGACAG GAGATCCTGC CCCGGCACTT CGCCCAATAG
  841 CAGCCAGTCC CTTCCCGCTT CAGTGACAAC GTCGAGCACA GCTGCGCAAG GAACGCCCGT
  901 CGTGGCCAGC CACGATAGCC GCGCTGCCTC GTCTTGCAGT TCATTCAGGG CACCGGACAG
  961 GGTCGGTCTT GACAAAAAGA ACCCGGGCGC CCCTGCGCTG ACAGCCCGGA ACACGGCGGC
 1021 ATCAGAGCAG CCGATTGTCT GTGTGCCCAG TCATAGCCCG ATAGCCTCTC CACCCAGGCG
 1081 GCTGAGAACT GCGTGCATCC ATCTTGGTCA TCATGCGAAC GATCCCTCAT CCTGTTCTTC
1141 TTTGGAAA
//

Please visit Sequencing and PCR primers for primer information.


References

References

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 .

2018.12.11

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