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pMK293 (mAID-mCherry2-Hygro)

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

Catalog number RDB13936
Resource name pMK293 (mAID-mCherry2-Hygro)
Alternative name mAID-mCherry2-Hygro, 977
Clone info. A plasmid for construction of C-terminal mAID-mCherry tagging donors. Please refer Fig. 4 of Natsume, T., Cell Reports 15, 210-218 (2016) for an instruction about how to use this clone.
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 Discosoma sp. RFP 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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Remarks An additional license fee for DNA resources containing fluorescent proteins owned by, or licensed to, Clontech Laboratories, Inc. is required.
The BIOLOGICAL RESOURCE may not be used for COMMERCIAL PURPOSES. Please read the Exclusive MTA (Clontech FPs).
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RDB13936 pMK293 (mAID-mCherry2-Hygro) 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 RDB13936_A6B8p1_v2.pdf

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

Primer: M13_-40 RDB13936_A6B8a_2.seq
>D03070A20_A6B8_1_M13(-40)_E05_14.ab1
    1 CGCGGGACGC GCGTATACGA CTCACTATAG GGCGAATTGG AGCTCCCCGG GATCCGGTGC
   61 AGGCGCCAAG GAGAAGAGTG CTTGTCCTAA AGATCCAGCC AAACCTCCGG CCAAGGCACA
  121 AGTTGTGGGA TGGCCACCGG TGAGATCATA CCGGAAGAAC GTGATGGTTT CCTGCCAAAA
  181 ATCAAGCGGT GGCCCGGAGG CGGCGGCGTT CGTGAAGGTA TCAATGGACG GAGCACCGTA
  241 CTTGAGGAAA ATCGATTTGA GGATGTATAA AGCTAGCATG GTGAGCAAGG GCGAGGAGGA
  301 TAACATGGCC ATCATCAAGG AGTTCATGCG CTTCAAGGTG CACATGGAGG GCTCCGTGAA
  361 CGGCCACGAG TTCGAGATCG AGGGCGAGGG CGAGGGCCGC CCCTACGAGG GCACCCAGAC
  421 CGCCAAGCTG AAGGTGACCA AGGGTGGCCC CCTGCCCTTC GCCTGGGACA TCCTGTCCCC
  481 TCAGTTCATG TACGGCTCCA AGGCCTACGT GAAGCACCCC GCCGACATCC CCGACTACTT
  541 GAAGCTGTCC TTCCCCGAGG GCTTCAATTG GGAGCGCGTG ATGAACTTCG AGGACGGCGG
  601 CGTGGTGACC GTGACCCAGG ACTCCTCCCT GCAGGACGGC GAGTTCATCT ACAAGGTGAA
  661 GCTGCGCGGC ACCAACTTCC CCTCCGACGG CCCCGTAATG CAGTGTCGTA CCATGGGCTG
  721 GGAGGCCTCC ACTGAGCGGA TGTACCCCGA GGACGGCGCC CTGAAGGGCG AGATCAAGCA
  781 GAGGCTGAAG CTGAAGGACG GCGGCCACTA CGACGCTGAG GTCAAGACCA CCTACAAGGC
  841 CAAGAAGCCC GTGCAGCTGC CCGGCGCCTA CAACGTCGAC ATCAAGTTGG ACATCCTTTT
  901 CCCACAACGA GGACTACACC ATCGTGGAAA CAGTACGAAC GCGCCGAGGG CCGCCACTCC
  961 CACCGGGCGG CATGGACGAG CTGTACAAGT AATCTAGATA ACTGATCATA ATCAGCCATA
 1021 CCACATTTGT AGAGGGTTTT TACTTTGCTT TAAAAAACTC CCACACCTCC CCCTGACCTG
 1081 AAACATAAAA TGATGCAATT GAGTGTACCT GGTTAATTGC AGCCTAATAA TGGATTACAA
1141 AATTAAAGGC AATTAG
//
Primer: PGK_N_rev RDB13936_A6B8b_2.seq
>D03070A20_A6B8_1_PGK-N-rev_G06_21.ab1
    1 CATGGACGCT CCCTACCGGT AGATTTCGAG GTCGAGATCC TAAGCTTGGC TGGACGTAAA
   61 CTCCTCTTCA GACCTAATAA CTTCGTATAG CATACATTAT ACGAAGTTAT ATTAAGGGTT
  121 ATTGAATATG ATCGACGCGT TAAGATACAT TGATGAGTTT GGACAAACCA CAACTAGAAT
  181 GCAGTGAAAA AAATGCTTTA TTTGTGAAAT TTGTGATGCT ATTGCTTTAT TTGTAACCAT
  241 TATAAGCTGC AATAAACAAG TTAACAACAA CAATTGCATT CATTTTATGT TTCAGGTTCA
  301 GGGGGAGGTG TGGGAGGTTT TTTAAAGCAA GTAAAACCTC TACAAATGTG GTATGGCTGA
  361 TTATGATCAG TTATCTAGAT TACTTGTACA GCTCGTCCAT GCCGCCGGTG GAGTGGCGGC
  421 CCTCGGCGCG TTCGTACTGT TCCACGATGG TGTAGTCCTC GTTGTGGGAA AGGATGTCCA
  481 ACTTGATGTC GACGTTGTAG GCGCCGGGCA GCTGCACGGG CTTCTTGGCC TTGTAGGTGG
  541 TCTTGACCTC AGCGTCGTAG TGGCCGCCGT CCTTCAGCTT CAGCCTCTGC TTGATCTCGC
  601 CCTTCAGGGC GCCGTCCTCG GGGTACATCC GCTCAGTGGA GGCCTCCCAG CCCATGGTAC
  661 GACACTGCAT TACGGGGCCG TCGGAGGGGA AGTTGGTGCC GCGCAGCTTC ACCTTGTAGA
  721 TGAACTCGCC GTCCTGCAGG GAGGAGTCCT GGGTCACGGT CACCACGCCG CCGTCCTCGA
  781 AGTTCATCAC GCGCTCCCAA TTGAAGCCCT CGGGGAAGGA CAGCTTCAAG TAGTCGGGGA
  841 TGTCGGCGGG GTGCTTCACG TAGGCCTTGG AGCCGTACAT GAACTGAAGG GGACAGGATG
  901 TCCCAGGCGA AGGCAGGGGG CCCACCTTGG GTCACCTTCA GCTTGGCGGT CTGGGTGCCC
  961 TCGTAAGGGC GGCCCTCGCC CCTCGCCCTC GAATCTCGAA CTCGTGGCCC GTTCACGGAG
 1021 CCCTCATGTG CACCTTGAAG CCGCATGGAA CCTCCCTGGA TGATGTGACC ATGTTATCCA
 1081 TCATCAGCCT TGCTTCACCC ATGGCTAGCC CTTATAGCAT CTCGAATTCG AATTCTTCAA
1141 GTACGGTCTC GTTCCATTGG ATACCATCAA CT
//
Primer: PGKpro_F RDB13936_A6B8c_2.seq
>D03070A20_A6B8_1_PGK-proF_B08_05.ab1
    1 ACCCGCGATC TCTCTTCTCA TCTCCGGGCC TTTCGACCTG CATCCATCTA GATCTCGAGC
   61 AGCTGAAGCT TACCGCTAGC ATGGATAGAT CCGGAAAGCC TGAACTCACC GCGACGTCTG
  121 TCGAGAAGTT TCTGATCGAA AAGTTCGACA GCGTCTCCGA CCTGATGCAG CTCTCGGAGG
  181 GCGAAGAATC TCGTGCTTTC AGCTTCGATG TAGGAGGGCG TGGATATGTC CTGCGGGTAA
  241 ATAGCTGCGC CGATGGTTTC TACAAAGATC GTTATGTTTA TCGGCACTTT GCATCGGCCG
  301 CGCTCCCGAT TCCGGAAGTG CTTGACATTG GGGAATTCAG CGAGAGCCTG ACCTATTGCA
  361 TCTCCCGCCG TGCACAGGGT GTCACGTTGC AAGACCTGCC TGAAACCGAA CTGCCCGCTG
  421 TTCTGCAGCC GGTCGCGGAG GCCATGGATG CGATCGCTGC GGCCGATCTT AGCCAGACGA
  481 GCGGGTTCGG CCCATTCGGA CCGCAAGGAA TCGGTCAATA CACTACATGG CGTGATTTCA
  541 TATGCGCGAT TGCTGATCCC CATGTGTATC ACTGGCAAAC TGTGATGGAC GACACCGTCA
  601 GTGCGTCCGT CGCGCAGGCT CTCGATGAGC TGATGCTTTG GGCCGAGGAC TGCCCCGAAG
  661 TCCGGCACCT CGTGCACGCG GATTTCGGCT CCAACAATGT CCTGACGGAC AATGGCCGCA
  721 TAACAGCGGT CATTGACTGG AGCGAGGCGA TGTTCGGGGA TTCCCAATAC GAGGTCGCCA
  781 ACATCTTCTT CTGGAGGCCG TGGTTGGCTT GTATGGAGCA GCAGACGCGC TACTTCGAGC
  841 GGAGGCATCC GGAGCTTGCA GATCGCCGCG GCTCCGGGCG TATATGCTCC GCATTGGTCT
  901 TGACCAACTC TATCAGAGCT TGGTTGACGG CAATTTCGAT GATGCAGCTT GGGCGCAGGG
  961 TCGATGCGAC GCAATCGTCC CGATCCGGAG CCGGGACTGT CGGGGCGTAC ACAAATCGCC
 1021 GCAGAAGCGC GGCCGTCTGG ACGATGGCTG TGTAGAGTAC TTCGCGATAG TGGAACGACG
 1081 CTCAGCACTC GTCCGAGCAA AGATTAGCTA GCCGGCCGTC CCCCACGACC CCGCCCAGTC
1141 G
//
Primer: M13R RDB13936_A6B8d_2.seq
>D03070A20_A6B8_1_M13R_E04_13.ab1
    1 CTCGCGCATT AACCTCACTA AAGGGACAAA AGCTGGGTAC CGGATCCTAG TGAACCTCTT
   61 CGAGGGACCT AATAACTTCG TATAGCATAC ATTATACGAA GTTATATTAA GGGTTCCGGA
  121 TCTCGACCAG CTTCTGATGG AATTAGAACT TGGCAAAACA ATACTGAGAA TGAAGTGTAT
  181 GTGGAACAGA GGCTGCTGAT CTCGTTCTTC AGGCTATGAA ACTGACACAT TTGGAAACCA
  241 CAGTACTTAG AACCACAAAG TGGGAATCAA GAGAAAAACA ATGATCCCAC GAGAGATCTA
  301 TAGATCTATA GATCATGAGT GGGAGGAATG AGCTGGCCCT TAATTTGGTT TTGCTTGTTT
  361 AAATTATGAT ATCCAACTAT GAAACATTAT CATAAAGCAA TAGTAAAGAG CCTTCAGTAA
  421 AGAGCAGGCA TTTATCTAAT CCCACCCCAC CCCCACCCCC GTAGCTCCAA TCCTTCCATT
  481 CAAAATGTAG GTACTCTGTT CTCACCCTTC TTAACAAAGT ATGACAGGAA AAACTTCCAT
  541 TTTAGTGGAC ATCTTTATTG TTTAATAGAT CATCAATTTC TGCATCCCGG GGATCTGATA
  601 TCATCGATGC ATGGGGTCGT GCGCTCCTTT CGGTCGGGCG CTGCGGGTCG TGGGGCGGGC
  661 GGCTAGCCTA TTCCTTTGCC CTCGGACGAG TGCTGGGGCG TCGGTTTCCA CTATCGGCGA
  721 GTACTTCTAC ACAGCCATCG GTCCAGACGG CCGCGCTTCT GCGGGCGATT TGTGTACGCC
  781 CGACAGTCCC GGCTCCGGAT CGGACGATTG CGTCGCATCG ACCCTGCGCC CAAGCTGCAT
  841 CATCGAAATT GCCGTCAACC AAGCTCTGAT AGAGTTGGTC AAGACCAATG CGGAGCATAT
  901 ACGCCCGGAG CCGCGGCGAT CCTGCAAGCT CCGGATGCCT CCGCTCGAAG TAGCGCGTCT
  961 GCTGCTCCAT ACAAGCCCAA CCCACGGGCC TCCAGAAGAA GATGTTGGCG ACCTCGTATT
 1021 GGGGAATCCC CGAACATCGC TCGCTCAGTC ATGAACCGCT GTATGCCGGC ATTGTCGTCA
1081 GGGACATGAT GAGCGGAATT CGCGTGCACG AGTGCCGGAA CCTTCCGGGG GGATCA
//

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 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 Tungadi, E.A., Human microcephaly ASPM protein is a spindle pole-focusing factor that functions redundantly with CDK5RAP2. J. Cell Sci. 130 (21): 3676-3684 (2017). PMID 28883092.
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 .

2019.03.08

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