Sequencing and PCR primers for primer information.
Expression clone for an auxin-based degron (AID) system, GFP-aid-NLS.
| Catalog number | RDB08468 |
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| Resource name | pNHK60 |
| Alternative name | OsTIR1-9Myc::IRES::GFP-aid-NLS (kan/neoR) |
| Clone info. | Expression clone for an auxin-based degron (AID) system. GFP-aid-NLS. Refer Supplementary figure 10 (pIRES OsTIR1-EGFP-IAA17) of Nat. Methods, 6, 917-922, 2009; PMID: 19915560. Please note that (1) the degron system may not work well in T-antigen expressing cells such as HEK293T because pNHK60 has the SV40 replication origin and the expression level of GFP-degron is quite high, (2) the depositor found that degradation is efficient in stable cells rather than in transiently transiently transfected cells. Accession numbers, OsTIR1 (GeneID 4335696); AtIAA17 (GeneID 839568). 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 | pIRES-EGFP (Plasmid) |
| Selectable markers | Kan^r |
| Gene/insert name | Oryza sativa OsTIR1 cDNA Arabidopsis thaliana AtIAA17 cDNA Aequorea victoria GFP cDNA |
| Depositor | Kanemaki, Masato | |
| Other clones in our bank | Oryza sativa LOC4335696 | Arabidopsis thaliana AXR3 (KEGG orthology K14484) | |
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(s) designated by the DEPOSITOR is requested (Nishimura, K., Nat. Methods., 6, 917-922 (2009)). 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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| Ordering | Please visit Information of Request for Distribution. Order form [Credit Card Payment] [Bank Transfer or Check Payment] Material Transfer Agreement (MTA) [Word] Schedule A (GFP-A) [Link] |
| Remarks | ((Additional form)) GFP-A Please use Schedule A (GFP-A) additionally. |
[open/close]| Catalog # | Resource name | Shipping form | Fee (non-profit org.) |
|---|---|---|---|
| RDB08468 | pNHK60 | DNA solution |
Electronic file
| Electronic file | Sequence RDB08468z.seq from Depositor |
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| Electronic file | PDF RDB08468.pdf from Depositor |
Featured content
| Featured content | Auxin Inducible Degron (AID) System (English text) |
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| Featured content | Auxin Inducible Degron (AID) System (Japanese text) |
References
| original | Nishimura, K., An auxin-based degron system for the rapid depletion of proteins in nonplant cells. Nat Methods. 6(12), 917-922, (2009). PMID 19915560. |
|---|---|
| original | Kanemaki, M., Method for inducing degradation of protein in mammalian cell WO2010125620A1 |
User reports and related articles (go to bottom)
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 | RDB08468_A5Dep1.pdf |
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Nucleotide sequence of a portion of this resource (if available).
| Primer: CMV-Forward | RDB08468_A5Dea.seq |
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>08468_08468_A5De_CMV-Forward_G08_20.ab1 1 CATCAACTCC GCCCCATTGA CGCAATGGGC GGTAGGCGTG TACGGTGGGA GGTCTATATA 61 AGCAGAGCTG GTTTAGTGAA CCGTCAGATC CGCTAGCGCT ACCGGACTCA GATCTCGAGA 121 TGACGTACTT CCCGGAGGAG GTGGTGGAGC ACATCTTCAG CTTCCTGCCG GCGCAGCGCG 181 ACCGCAACAC GGTCTCGCTC GTCTGCAAGG TGTGGTACGA GATCGAGAGG CTGAGCCGCC 241 GCGGCGTCTT CGTGGGCAAC TGCTACGCCG TGCGCGCCGG CCGCGTCGCC GCGCGGTTCC 301 CCAACGTGCG GGCGCTCACG GTGAAGGGGA AGCCCCACTT CGCCGACTTC AACCTCGTGC 361 CCCCCGACTG GGGCGGCTAC GCGGGGCCGT GGATCGAGGC GGCCGCGAGG GGATGCCACG 421 GCCTGGAGGA GCTCAGGATG AAGCGGATGG TGGTGTCCGA CGAGAGCCTC GAGCTGCTGG 481 CTCGCTCGTT CCCGCGGTTC AGGGCTCTTG TTCTTATCAG CTGCGAGGGG TTCAGCACTG 541 ACGGGCTAGC CGCCGTCGCG AGCCATTGCA AGCTTCTGAG GGAGTTGGAT TTGCAGGAAA 601 ATGAAGTGGA GGATCGAGGG CCTAGGTGGC TTTCCTGCTT CCCTGATTCC TGCACATCAC 661 TTGTCTCATT GAATTTTGCC TGCATCAAAG GGGAGGTTAA TGCTGGTTCA CTGGAGAGAC 721 TTGTTAGCAG GTCCCCAAAC CTGCGGAGTT TGAGGCTGAA TCGATCTGTA TCGGTAGATA 781 CACTTGCAAA GATACTACTG CGTACCCCTA ACTTGGAGGA TTTGGGGACA GGGAATTTGA 841 CAGATGACTT CCAAACTGAG TCCTACTTTA AGCTTACCAG TGCTCTGGAG AAATGCAAGA 901 TGTTGAGGAG TTTGTCTGGA TTCTGGGATG CTTCTCCTGT TTGCCTGTCA TTTATCTACC 961 CCCCTGTGTG CTCAACTGAC AGGATTGAAC TTGAGCTATG CACCCACACT TGATGCTTCT 1021 GACCCTTACA AAAATGATTA GCCGCTGTGT GAAGCTCCAA CGCCTTTGGG TACTGGATGT 1081 ATCTCGGGAC AAGGGCTGCA GTTGATTGCC TCAAGCTTGC AAGACTTTGC AAGAACTCTC 1141 AGGGATATTT TTGCAT // |
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| Primer: ME-1290Rv | RDB08468_A5Deb.seq |
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>08468_08468_A5De_ME-1290Rv_E04_13.ab1 1 ATGCAATTCA ATTTTATGTT TCAGGTTCAG GGGGAGGTGT GGGAGGTTTT TTAAAGCAAG 61 TAAAACCTCT ACAAATGTGG TATGGCTGAT TATGATCTAG AGTCGCGGCC GCTGGGTACC 121 TTAAACCTTA CGTTTCTTTT TAGGGACCTT TCTTTTCTTC TTGGGGGCGC CAGCACCAGC 181 TCTGCTCTTG CACTTCTCCA TCGCCCTCGG AGCGAGACCA ATGGCATCCG ATCCTTTCAT 241 GAGACGTAAA CGCTTGCATG TATCGACGAA CATTGGCCAA GGAACGTCGC CGACGAGCAT 301 CCAATCACCG TCTTTGTCTT CATAAGAGGG AACATAGTCC CAGCTATTCA CCAAATCCAT 361 CAATTTCCTC TCATTCATGA AGTCTATCAT TCCTTCTTCT CCTCCATGTT TGCCCATGGT 421 AAAAGAGCTG AACATGTTGG ACAAAGCATT AGAAAGCTCA TCGTAGCTTT TATACATCCT 481 CAAATCGATT TTCCTCAAGT ACGGTGCTCC GTCCATTGAT ACCTTCACGA ACGCCGCCGC 541 CTCCGGGCCA CCGCTTGATT TTTGGCAGGA AACCATCACG TTCTTCCGGT ATGATCTCAC 601 CGGTGGCCAT CCCACAACTT GTGCCTTGGC CGGAGGTTTG GCTGGATCTT TAGGACAAGC 661 ACTCTTCTCC TTGGAATCAA AAGTCACGAC GTCATGAGTC GTAGATCCTT CCTTGTTTGC 721 AGGCTCATTA TTCAGATTTA GCTTCAGATC AACCGTCTCT GAGAACCCTC TCTTGTTTCC 781 GGTTACCGGA GCCACTGTAT CTCCACCGGG AAGACCAAGA CACAGCTCAG TCTCCCTCAG 841 ATTCAGCTCG ACACTGCCCA TCATGGGCCC GGCACCCGCT CCAGCGCCTG CACCAGCTCC 901 CTTGTACAGC TCGTCCATGC CGAGAGTGAT CCCGGCGGCG GTCACGAACT CCAGCAGGAC 961 CATGTGATCG CGCTTCTCGT TGGGGTCTTT TGCTCAGGCG GACTGGTGCT CAGGTAGTGG 1021 TTTGTCGGGC AGCAGCACGG GGTCGTCGCG ATGGGGGGTG TTTCTGCTGT AGTGATCGGC 1081 GAGCTGCACG CTGCCGTCTT CGAATGGTGT GGCCGGAATC CTTGAGGTTC ACCCTTTGAT 1141 GACCGGAT // |
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| Primer: IRES-F | RDB08468_A5Dec.seq |
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>08468_08468_A5De_IRES-F_A09_03.ab1 1 CGTTCCCTTT AATGTGTTTA GTCGAGGTTA AAAACGTCTA GGCCCCCCGA ACCACGGGGA 61 CGTGGTTTTC CTTTGAAAAA CACGATGATA ATATGGCCAC AACCATGGTG AGCAAGGGCG 121 AGGAGCTGTT CACCGGGGTG GTGCCCATCC TGGTCGAGCT GGACGGCGAC GTAAACGGCC 181 ACAAGTTCAG CGTGTCCGGC GAGGGCGAGG GCGATGCCAC CTACGGCAAG CTGACCCTGA 241 AGTTCATCTG CACCACCGGC AAGCTGCCCG TGCCCTGGCC CACCCTCGTG ACCACCCTGA 301 CCTACGGCGT GCAGTGCTTC AGCCGCTACC CCGACCACAT GAAGCAGCAC GACTTCTTCA 361 AGTCCGCCAT GCCCGAAGGC TACGTCCAGG AGCGCACCAT CTTCTTCAAG GACGACGGCA 421 ACTACAAGAC CCGCGCCGAG GTGAAGTTCG AGGGCGACAC CCTGGTGAAC CGCATCGAGC 481 TGAAGGGCAT CGACTTCAAG GAGGACGGCA ACATCCTGGG GCACAAGCTG GAGTACAACT 541 ACAACAGCCA CAACGTCTAT ATCATGGCCG ACAAGCAGAA GAACGGCATC AAGGTGAACT 601 TCAAGATCCG CCACAACATC GAGGACGGCA GCGTGCAGCT CGCCGACCAC TACCAGCAGA 661 ACACCCCCAT CGGCGACGGC CCCGTGCTGC TGCCCGACAA CCACTACCTG AGCACCCAGT 721 CCGCCCTGAG CAAAGACCCC AACGAGAAGC GCGATCACAT GGTCCTGCTG GAGTTCGTGA 781 CCGCCGCCGG GATCACTCTC GGCATGGACG AGCTGTACAA GGGAGCTGGT GCAGGCGCTG 841 GAGCGGGTGC CGGGCCCATG ATGGGCAGTG TCGAGCTGAA TCTGAGGGAG ACTGAGCTGT 901 GTCTTGGTCT TCCCGGTGGA GATACAGTGG CTCCGGTAAC CGGAAACAAG AGAGGGTTCT 961 CAGAGACGGT TGATCTGAAG CTAAATCTGA ATAATGAGCC TGCAAACAAG GAAAGGATCT 1021 ACGACTCATG ACGTCGTGAC TTTTGATTCC AAGGAGAGGA GTGCTTTGTC CTAAAGATCC 1081 AGCAATCTTC GGGCCAAGGC ACAAGTTGTG GGATGCACGA TGAGATCATA CCGGAAGAAC 1141 GTGATGATTT CCTGGCCCAG A // |
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| Primer: IRES-R | RDB08468_A5Ded.seq |
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>08468_08468_A5De_IRES-R_B09_06.ab1 1 GCAAAAACCG GGCCTTCGGC CAGTAACGTT AGGGGGGGGG GAGGGAGAGG GGCGGATCCC 61 GGGGATCTCT TGAATGATCG TTCCACTTTT TAGCTAGTGG ATCCGTTCAA GTCTTCTTCT 121 GAGATTAATT TTTGTTCACC GTTCAAGTCT TCCTCGGAGA TTAGCTTTTG TTCACCGTTC 181 AAATCTTCTT CAGAAATCAA CTTTTGTTCA CCTCTAGAGG ATCCGTTCAA GTCTTCTTCT 241 GAGATTAATT TTTGTTCACC GTTCAAGTCT TCCTCGGAGA TTAGCTTTTG TTCACCGTTC 301 AAATCTTCTT CAGAAATCAA CTTTTGTTCA CCTCTAGAGG ATCCGTTCAA GTCTTCTTCT 361 GAGATTAATT TTTGTTCACC GTTCAAGTCT TCCTCGGAGA TTAGCTTTTG TTCACCGTTC 421 AAATCTTCTT CAGAAATCAA CTTTTGTTCA CCACTAGCAG CAGAACCGGA GTCGACTAGG 481 ATTTTAACAA AATTTGGTGC ATCATCCCTC GCCCCAGCAG TTGTGCGGTA CACATATAAT 541 TTCTCCACTT TAGGCAGATC TCCATGGTTT TCCTCCATTT CATTGCTACC ATCCCGCTCA 601 TTTATGACCT CAACATTGAG CATCGGCATC TTTGACGCAA GGACTTGGCA CCCCTTTAAC 661 GTGACATTGC AAGATGACAT CCAAAGGGAT CGCATTGTCT CGTACCTAGC AAAATTCCCC 721 AAGAGTGCAG CATCACCAAA CGGGCTATCT CTTATCTCCA GTTTCCTTAA ATTCTTGCAT 781 CCATTCATAA CATGCATCAT ACCCTTATCA CTGTCACCAG CAAATGCTAT AGAAAGCATC 841 TCAAGTTGTT TTGCATATTT CCCAATATAC ATGAAAACTT TGTCTGTGAG AAGACCAGAT 901 ATTGACAAAC GTTGTAATCC CTTGCACTCA CGAACAATAG CTCCAAAGCC TTCATCTAAT 961 GGTTGGCTTG TCACAACATC AGGCTTCCCT GGCTCAAGAA TACAAAGTCT GAATCGTGTG 1021 AAATTTGGAC AGTTCTTGGC GACAGTAACT AGTGCAGCAT TAGTCATTTG GTGACAGAAG 1081 TACAGTAGTG AGTCAGTTTT GGACAGCCAG GATACTGCAC AGGTTCCTTC GTCTGGTCAC 1141 TGCCAGATAC CCAGCTACGT TAAGAAAATC TCTGAATGTG // |
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| Primer: SV40pro_F_V2 | RDB08468_A5Dee.seq |
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>08468_08468_A5De_SV40proF_V2_F04_16.ab1 1 GACGGGCTTT TTTGGAGGCC 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 GCCCAACCTG CCATCACGAG ATTTCGATTC CACCGCCGCC TTCTATGAAG GTTGGGGCTT 961 CGGAATCGTT TTCCGGGACG CCGGCTGGAT GATCCTCCAG CGCGGGGATC TCATGCTGGA 1021 GTTCTTTCGC CCACCCTAGG GGGAGGCTAA CTGAAACACG GAAGGAGACA TACCGGAGAC 1081 CCGCGCTATG ACGGCATTAA AAGACCGATT AAACGCACGG TGATGGGATC GGATTGGTTC 1141 ATACGCGGAG TCGGATTCCA GGATCTGTGG CA // |
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Please visit Sequencing and PCR primers for primer information.
References
| original | Nishimura, K., An auxin-based degron system for the rapid depletion of proteins in nonplant cells. Nat Methods. 6(12), 917-922, (2009). PMID 19915560. |
|---|---|
| original | Kanemaki, M., Method for inducing degradation of protein in mammalian cell WO2010125620A1 |
| 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 | Delacour, Q., Light-Activated Proteolysis for the Spatiotemporal Control of Proteins. ACS Chem. Biol.10 (7): 1643-1647 (2015). PMID 25938742. |
| reference | Wend, S., A quantitative ratiometric sensor for time-resolved analysis of auxin dynamics. Sci. Rep. 3: 2052 (2014). PMID 23787479. |
| reference | Farr, C.J., The alpha isoform of topoisomerase II is required for hypercompaction of mitotic chromosomes in human cells. Nucleic Acids Res. 42 (7): 4414-4426 (2014). PMID 24476913. |
| 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 . |
| reference | Kampf, M.M., Rewiring and dosing of systems modules as a design approach for synthetic mammalian signaling networks. Mol. Biosyst. 8 (6): 1824-1832 (2012). PMID 22532387. |
| reference | Holland, A.J., Inducible, reversible system for the rapid and complete degradation of proteins in mammalian cells. Proc. Natl. Acad. Sci. U.S.A. 109 (49): E3350-7 PMID 23150568. |
2018.12.10
GNP_filter3_RDBDEP_html_181012.pl