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pMK243 (TetOne OsTIR1 AAVS1)

A donor vector for introducing Tet-OsTIR1 at the human AAVS1 locus.

Catalog number RDB13919
Resource name pMK243 (TetOne OsTIR1 AAVS1)
Alternative name TetOne OsTIR1 AAVS1
Clone info. A donor vector for introducing Tet-OsTIR1 at the human AAVS1 locus. Please refer Fig. 3 of Natsume, T., Cell Reports 15, 210-218 (2016) for an instruction about how to use this clone.
Oryza sativa OsTIR1 (NP_001052659.1), codon optimized cDNA for the expression in human cells. 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 pCR4Blunt-TOPO (Plasmid)
Size of vector backbone 4.0 kb
Selectable markers Am^r (E. coli), Puro^r (mammalian cells)
Gene/insert name human AAVS1 genomic Oryza sativa OsTIR1 cDNA
Depositor|Developer Kanemaki, Masato |
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External Database
human AAVS1 Oryza sativa OsTIR1

        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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        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]
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        RDB13919 pMK243 (TetOne OsTIR1 AAVS1) DNA solution

        Ordering Information [in Japanese] [in English]
        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 RDB13919_A6B8p1-3.pdf check

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

        Primer: TetOne oligo 1
        Region: SV40 p(A), Tet-On 3G
        Sequence file: RDB13919_A6B8g.seq check
        >D03070A03_A6B8_1_Tet One oligo1_E09_15.ab1
            1 CTGCAGTGAA AAATGCTTTA TTTGTGAAAT TTGTGATGCT ATTGCTTTAT TTGTAACCAT
           61 TATAAGCTGC AATAAACAAG TTAACAACAA CAATTGCATT CATTTTATGT TTCAGGTTCA
          121 GGGGGAGGTG TGGGAGGTTT TTTAAAGCAA GTAAAACCTC TACAAATGTG GTATGGCTGA
          181 TTATGATCCT CTGGAGATTT ACCCGGGGAG CATGTCAAGG TCAAAATCGT CAAGAGCGTC
          241 AGCAGGCAGC ATATCAAGGT CAAAGTCGTC AAGGGCATCG GCTGGGAGCA TGTCTAAGTC
          301 AAAATCGTCA AGGGCGTCGG TCGGCCCGCC GCTTTCGCAC TTTAGCTGTT TCTCCAGGCC
          361 ACATATGATT AGTTCCAGGC CGAAAAGGAA GGCAGGTTCG GCTCCCTGCC GGTCGAACAG
          421 CTCAATTGCT TGTTTCAGAA GTGGGGGCAT AGAATCGGTG GTAGGTGTCT CTCTTTCCTC
          481 TTTTGCTACT TGATGCTCCT GTTCCTCCAA TACGCAGCCC AGTGTAAAGT GGCCCACGGC
          541 GGACAGAGCG TACAGTGCGT TCTCCAGGGA GAAGCCTTGC TGACACAGGA ACGCGAGCTG
          601 ATTTTCCAGG GTTTCGTACT GTTTCTCTGT TGGGCGGGTG CCGAGATGCA CTTTAGCCCC
          661 GTCGCGATGT GAGAGGAGAG CACAGCGGTA TGACTTGGCG TTGTTCCGCA GAAAGTCTTG
          721 CCATGACTCG CCTTCCAGGG GGCAGGAGTG GGTATGATGC CTGTCCAGCA TCTCGATTGG
          781 CAGGGCATCG AGCAGGGCCC GCTTGTTCTT CACGTGCCAG TACAGGGTAG GCTGCTCAAC
          841 TCCCAGCTTT TGAGCGAGTT TCCTTGTCGT CAGGCCTTCG ATACCGACTC CATTGAGTAA
          901 TTCCAGGAGG CAGAGTTTAT GACTTTGGCT TCTTGTCCAG TCTAGACATG GGTAATTCGA
          961 TGATCCCCCC TGGGGAGAGA GGTCGGTGAA TTCGGTCAAC GAAGGGAGTC GGACTGGCGG
         1021 ACGTGCGCCT TCGGAGCCTG CCAGATGCCG AAACACCGCG TCGGGCCAGA CATGGCTCAA
        1081 ACTACGACCA AGCCTGCTTC GGACCGGTCG TTCCAGGTCA GCTTAGCT
        //
        Primer: pause site
        Region: TRE3GS pro, OsTIR1 5'
        Sequence file: RDB13919_A6B8h.seq check
        >D03070A03_A6B8_1_Pause site_F09_18.ab1
            1 GATTCCTTTC TCTCTCGAGT TTACTCCCTA TCAGTGATAG AGAACGTATG AAGAGTTTAC
           61 TCCCTATCAG TGATAGAGAA CGTATGCAGA CTTTACTCCC TATCAGTGAT AGAGAACGTA
          121 TAAGGAGTTT ACTCCCTATC AGTGATAGAG AACGTATGAC CAGTTTACTC CCTATCAGTG
          181 ATAGAGAACG TATCTACAGT TTACTCCCTA TCAGTGATAG AGAACGTATA TCCAGTTTAC
          241 TCCCTATCAG TGATAGAGAA CGTATAAGCT TTGCTTATGT AAACCAGGGC GCCTATAAAA
          301 GAGTGCTGAT TTTTTGAGTA AACTTCAATT CCACAACACT TTTGTCTTAT ACCAACTTTC
          361 CGTACCACTT CCTACCCTCG TAAAGAATTC GCGGCCGCAC GCGTGCCACC ATGACATACT
          421 TTCCTGAAGA GGTCGTCGAA CACATTTTTA GCTTCCTGCC TGCACAGAGA GATAGAAACA
          481 CAGTGAGCCT GGTCTGCAAA GTGTGGTACG AGATCGAACG CCTGAGCCGG AGAGGAGTGT
          541 TCGTCGGCAA CTGCTATGCT GTGAGAGCAG GCAGGGTCGC CGCTAGGTTT CCAAATGTGC
          601 GCGCACTGAC CGTCAAGGGG AAACCCCACT TCGCCGACTT TAACCTGGTG CCCCCTGATT
          661 GGGGAGGATA CGCCGGCCCT TGGATCGAGG CAGCCGCTCG CGGCTGTCAT GGACTGGAGG
          721 AACTGCGCAT GAAGCGAATG GTGGTCTCTG ACGAAAGTCT GGAGCTGCTG GCTCGGAGCT
          781 TCCCTAGGTT TCGCGCACTG GTGCTGATTT CTTGCGAAGG CTTCAGCACC GATGGACTGG
          841 CAGCCGTGGC CTCCCACTGT AAGCTGCTGC GGGAGCTGGA CCTCCAGGAG AATGAAGTGG
          901 AGGATAGAGG CCCCAGATGG CTGTCTTGCT TCCCAGACTC ATGTACCAGC CTGGGTGTCC
          961 CTGAACTTTG CCTGCATCAA AGCGAAGTGA ATGCTGGGTC CCTGGAGCGG CTGGGTCTCA
         1021 AGAAGCCCCC ACCTGAGTCT CTGCCGCTGA ACCCGGAGCG TGGAGCGTGA ACACTCTGGC
        1081 TAGATCTGCT TGAGACCCCT TAACCTGCAG GATTCTGTGG GAAACCGC
        //
        Primer: BGH_rev2
        Region: BGH p(A), PuroR
        Sequence file: RDB13919_A6B8i.seq check
        >D03070A03_A6B8_1_BGH_rev2_G09_21.ab1
            1 CTTCGTAGAG CACGGGGGAG GGGCAACAAC AGATGGCTGG CAACTAGAAG GCACAGGGTA
           61 CCTCAGGCAC CGGGCTTGCG GGTCATGCAC CAGGTGCGCG GTCCTTCGGG CACCTCGACG
          121 TCGGCGGTGA CGGTGAAGCC GAGCCGCTCG TAGAAGGGGA GGTTGCGGGG CGCGGAGGTC
          181 TCCAGGAAGG CGGGCACCCC GGCGCGCTCG GCCGCCTCCA CTCCGGGGAG CACGACGGCG
          241 CTGCCCAGAC CCTTGCCCTG GTGGTCGGGC GAGACGCCGA CGGTGGCCAG GAACCACGCG
          301 GGCTCCTTGG GCCGGTGCGG CGCCAGGAGG CCTTCCATCT GTTGCTGCGC GGCCAGCCGG
          361 GAACCGCTCA ACTCGGCCAT GCGCGGGCCG ATCTCGGCGA ACACCGCCCC CGCTTCGACG
          421 CTCTCCGGCG TGGTCCAGAC CGCCACCGCG GCGCCGTCGT CCGCGACCCA CACCTTGCCG
          481 ATGTCGAGCC CGACGCGCGT GAGGAAGAGT TCTTGCAGCT CGGTGACCCG CTCGATGTGG
          541 CGGTCCCGGA TCGACGTGTG GCGCGTGGCG GGGTAGTCGG CGAACGCGGC GGCGAGGGTG
          601 CGTACGGCCC TGGGGACGTC GTCGCGGGTG GCGAGGGCGC ACCGTGGGCT TGTACTTCGG
        661 GTCATGGTAA GCTCCGGTGG ATCCCCCTTG
        //
        Primer: M13R
        Region: AAVS1 arm 1
        Sequence file: RDB13919_A6B8j.seq check
        >D03070A03_A6B8_1_M13R_F10_16.ab1
            1 AAATCTCAAA TTACCCCTCA CTAAAGGGAC TAGTCCTGCA GGTTTAAAAC GAATTCGCCC
           61 TTTGCTTTCT CTGACCAGCA TTCTCTCCCC TGGGCCTGTG CCGCTTTCTG TCTGCAGCTT
          121 GTGGCCTGGG TCACCTCTAC GGCTGGCCCA GATCCTTCCC TGCCGCCTCC TTCAGGTTCC
          181 GTCTTCCTCC ACTCCCTCTT CCCCTTGCTC TCTGCTGTGT TGCTGCCCAA GGATGCTCTT
          241 TCCGGAGCAC TTCCTTCTCG GCGCTGCACC ACGTGATGTC CTCTGAGCGG ATCCTCCCCG
          301 TGTCTGGGTC CTCTCCGGGC ATCTCTCCTC CCTCACCCAA CCCCATGCCG TCTTCACTCG
          361 CTGGGTTCCC TTTTCCTTCT CCTTCTGGGG CCTGTGCCAT CTCTCGTTTC TTAGGATGGC
          421 CTTCTCCGAC GGATGTCTCC CTTGCGTCCC GCCTCCCCTT CTTGTAGGCC TGCATCATCA
          481 CCGTTTTTCT GGACAACCCC AAAGTACCCC GTCTCCCTGG CTTTAGCCAC CTCTCCATCC
          541 TCTTGCTTTC TTTGCCTGGA CACCCCGTTC TCCTGTGGAT TCGGGTCACC TCTCACTCCT
          601 TTCATTTGGG CAGCTCCCCT ACCCCCCTTA CCTCTCTAGT CTGTGCTAGC TCTTCCAGCC
          661 CCCTGTCATG GCATCTTCCA GGGGTCCGAG AGCTCAGCTA GTCTTCTTCC TCCAACCCGG
          721 GCCCCTATGT CCACTTCAGG ACAGCATGTT TGCTGCCTCC AGAGATCCCT GTGTCCCCGA
        781 GCTGGGACCA ACCTT
        //
        Primer: M13_-40
        Region: T7 pro, AAVS1 arm 2
        Sequence file: RDB13919_A6B8k.seq check
        >D03070A03_A6B8_1_M13(-40)_G10_19.ab1
            1 CAGCCCGGAA TGTATACGAC TCACTATAGG GCGAATTGAA TTTAGCGGCC GCGAATTCGC
           61 CCTTAGAGCA GAGCCAGGAA CCCCTGTAGG GAAGGGGCAG GAGAGCCAGG GGCATGAGAT
          121 GGTGGACGAG GAAGGGGGAC AGGGAAGCCT GAGCGCCTCT CCTGGGCTTG CCAAGGACTC
          181 AAACCCAGAA GCCCAGAGCA GGGCCTTAGG GAAGCGGGAC CCTGCTCTGG GCGGAGGAAT
          241 ATGTCCCAGA TAGCACTGGG GACTCTTTAA GGAAAGAAGG ATGGAGAAAG AGAAAGGGAG
          301 TAGAGGCGGC CACGACCTGG TGAACACCTA GGACGCACCA TTCTCACAAA GGGAGTTTTC
          361 CACACGGACA CCCCCCTCCT CACCACAGCC CTGCCAGGAC GGGGCTGGCT ACTGGCCTTA
          421 TCTCACAGGT AAAACTGACG CACGGAGGAA CAATATAAAT TGGGGACTAG AAAGGTGAAG
          481 AGCCAAAGTT AGAACTCAGG ACCAACTTAT TCTGATTTTG TTTTTCCAAA CTGCTTCTCC
          541 TCTTGGGAAG TGTAAGGAAG CTGCAGCACC AGGATCAGTG AAACGCACCA GACAGCCGCG
          601 TCAGAGCAGC TCAGGTTCTG GGAGAGGGTA GCGCAGGGTG GCCACTGAGA ACCGGGCAGG
          661 TCACGCATCC CCCCCTTCCC TCCCACCCCC TGCCAAGCTC TCCCTCCCAG GATCCTCTCT
          721 GGCTCCATCG TAAGCAAACC TTAGAGGTTC TGGCAAGGAG AGAGATGGCT CCAGGAAATG
          781 GGGGTGTGTC ACCAGATAAG GAATCTGCCT AACAGGAGGT GGGGGTTAGA CCCAATATCA
          841 GGAGACTAGG AAAGGAGGAG GCCTAAGGAT GGGGCTTTTC TGTCACCAAT CCTGTCCCTA
          901 GTAAAGCTTG GTACCCCATA GAGCCCACCG CATCCCCAGC ATGCCTGCTA TTGTCTTCCC
          961 AATCCTCCCC CTTGCTGTCC TGCCCACCCA CCCCCAGAAT AAGAATGACA CCTACTCAGA
         1021 CAATGCGATG CAATTTTCTC ATTTATTTAG GAAAGGACAG TGGGAGTGGC ACTTCCAGGT
        1081 CCAGATGCAC TGGGAAGGTC AAACCACAGA ATGGCTTGCC AACCTTGAGG AAAGAGGCA
        //
        Primer: TetOne oligo 2
        Region: OsTIR1 3'
        Sequence file: RDB13919_A6B8l.seq check
        >D03070A03_A6B8_1_TetOne oligo 2_H10_22.ab1
            1 CAGGTGCAAA AAATGCTTTA TTTGTGAAAT TTGTGATGCT ATTGCTTTAT TTGTAACCAT
           61 TATAAGCTGC AATAAACAAG TTAACAACAA CAATTGCATT CATTTTATGT TTCAGGTTCA
          121 GGGGGAGGTG TGGGAGGTTT TTTAAAGCAA GTAAAACCTC TACAAATGTG GTATGGCTGA
          181 TTATGATCCT CTAGACATAT GCTGCAGAGA TCTTCACAGA ATCTTCACAA AGTTGGGAGC
          241 ATCATCTCTT GCCCCTGCGG TGGTCCGATA CACATACAGT TTCTCCACCT TTGGCAGGTC
          301 GCCATGATTT TCCTCCATTT CGTTAGACCC GTCCCGCTCA TTAATGACTT CCACGTTCAG
          361 CATAGGCATT TTACTAGCCA GGACCTGACA GCCCTTCAGA GTCACATTGC AACTAGACAT
          421 CCACAGGCTC CTCATTGTCT CGTATCTGGC AAAGTTTCCC AGCAGAGCGG CATCCCCGAA
          481 AGGGCTGTCC CGGATTTCCA GTTTTCGCAG ATTCTTACAC CCCGTTCATC ACGTGCATCA
          541 TGCCCTTATC GGAGTCTCCG GCAAATGCAT GGTTCACATC TCAGCTGCTT AGCATACTTG
          601 CCGATGTACA TGAACACTTT TGTCTGTCAG CAGGCCGGAA ATGCTAAGCC TCTGAGATTC
        661 CTTGC
        //

        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 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.

        2022.02.09

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