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

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.
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 Discosoma sp. RFP cDNA
Depositor|Developer Kanemaki, Masato |
Other clones in our bank

External Database
Arabidopsis thaliana AtIAA17 Discosoma sp. RFP

          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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          This RESOURCE contains a fluorescent protein owned or licensed by Takara Bio USA, Inc. (TBUSA). Please refer to Clontech Limited Use Label License and use Exclusive MTA (download the LULL and the MTA [link]).
          Remarks An additional license fee for DNA resources containing fluorescent proteins owned by, or licensed to, Clontech Laboratories, Inc. is required.
          This RESOURCE contains a fluorescent protein owned or licensed by Takara Bio USA, Inc. A part of license fees is required. This RESOURCE is not provided to for-profit organization or for for-profit research by non-profit organizations.
          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]
          Takara Bio USA, Inc.が保有している蛍光タンパク質を含みます。 Clontech Limited Use Label Licenseをご一読の上専用MTAの提出をお願いします (専用MTA (英語版のみ) とClontech Limited Use Label Licenseを入手[link])。
          備考 クロンテック社の蛍光タンパク質 (DsRed2、mCherry)を含むクローン。ライセンス料相当額を負担して頂きます。
          Takara Bio USA, Inc.が保有している蛍光タンパク質を含みます。ライセンス料相当額を負担して頂きます。本件リソースは営利機関および非営利機関による営利目的研究には提供いたしません。

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          RDB13936 pMK293 (mAID-mCherry2-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 RDB13936_A6B8p1-2 check

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

          Primer: M13_-40
          Region: Linker, mAID, mCherry2 5'
          Sequence file: RDB13936_A6B8e.seq check
          >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
          Region: loxP, SV40 p(A), mCherry2 3'
          Sequence file: RDB13936_A6B8f.seq check
          >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
          Region: PGK pro, HygR 5'
          Sequence file: RDB13936_A6B8g.seq check
          >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
          Region: loxP, mouse PGK ter, HygR 3'
          Sequence file: RDB13936_A6B8h.seq check
          >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

          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 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.

          2022.05.17

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