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

pMK232 (AAVS1 CMV-OsTIR1-PURO)

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

Catalog number RDB13918
Resource name pMK232 (AAVS1 CMV-OsTIR1-PURO)
Alternative name AAVS1 CMV-OsTIR1-PURO
Clone info. A donor vector for introducing CMV-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.
Comment 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]
        提供同意書 (MTA、非営利機関による非営利学術研究用)[Word]

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        RDB13918 pMK232 (AAVS1 CMV-OsTIR1-PURO) 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 RDB13918_A6B8p1-3.pdf check

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

        Primer: CMV_Forward
        Region: insert mid (CMV pro, OsTIR1 5')
        Sequence file: RDB13918_A6B8f.seq check
        >D03070A02_A6B8_1_CMV_Forward_D10_10.ab1
            1 ACGTCACTCC GCCCATTGAC GCAATGGGCG GTAGGCGTGT ACGGTGGGAG GTCTATATAA
           61 GCAGAGCTGG TTTAGTGAAC CGTCAGATCC GCTAGCGCTG CCACCATGAC ATACTTTCCT
          121 GAAGAGGTCG TCGAACACAT TTTTAGCTTC CTGCCTGCAC AGAGAGATAG AAACACAGTG
          181 AGCCTGGTCT GCAAAGTGTG GTACGAGATC GAACGCCTGA GCCGGAGAGG AGTGTTCGTC
          241 GGCAACTGCT ATGCTGTGAG AGCAGGCAGG GTCGCCGCTA GGTTTCCAAA TGTGCGCGCA
          301 CTGACCGTCA AGGGGAAACC CCACTTCGCC GACTTTAACC TGGTGCCCCC TGATTGGGGA
          361 GGATACGCCG GCCCTTGGAT CGAGGCAGCC GCTCGCGGCT GTCATGGACT GGAGGAACTG
          421 CGCATGAAGC GAATGGTGGT CTCTGACGAA AGTCTGGAGC TGCTGGCTCG GAGCTTCCCT
          481 AGGTTTCGCG CACTGGTGCT GATTTTCTTG CGAAAGGCTT CAGCACCGAT GGACTGGCAG
          541 CCGTGGCCTC CCACTGTAAG CTGCTGCGGG GAGCTGGACC TTCCAGGAAG AATGGAGTGG
          601 AGGATAGAGG GCCCAAATGG CTGTCTTGCT TCCCAGACTC ATGTACCAGC CTTGGTGTTC
          661 CCTGAACTTT GCCTGCATCA AAGGCGAAAT GATGGCTGGG TCCCTGGGAC CGGCTGGGTT
          721 TTCAAGAAGC CCACACTTGA AGGTCTCTTG CGGCTGGAAC CGGGAGCCTG GAGCGTGGAA
          781 CACTCTGGGC TAAAATTCTG CTGGAGAACC CTTAACCTGG AGAATCTGGG AACGGCAATC
          841 TCGACAGACG AATTTGGACA GGAGTCTACT ATTTTGCCTT CGGCTTTGGA AGTCAATGGC
        901 GGGAGAGGGT CTCAGAAATC CGTGGAGTCT CACACCGGTT GTCC
        //
        Primer: BGH_rev2
        Region: insert mid (bGH p(A), PuroR,hPGK pro)
        Sequence file: RDB13918_A6B8g.seq check
        >D03070A02_A6B8_1_BGH_rev2_E10_13.ab1
            1 CATCCGCGGT CAGGAGGCAC GGGGGAGGGG CAAACAACAG ATGGCTGGCA ACTAGAAGGC
           61 ACAGGGTACC TCAGGCACCG GGCTTGCGGG TCATGCACCA GGTGCGCGGT CCTTCGGGCA
          121 CCTCGACGTC GGCGGTGACG GTGAAGCCGA GCCGCTCGTA GAAGGGGAGG TTGCGGGGCG
          181 CGGAGGTCTC CAGGAAGGCG GGCACCCCGG CGCGCTCGGC CGCCTCCACT CCGGGGAGCA
          241 CGACGGCGCT GCCCAGACCC TTGCCCTGGT GGTCGGGCGA GACGCCGACG GTGGCCAGGA
          301 ACCACGCGGG CTCCTTGGGC CGGTGCGGCG CCAGGAGGCC TTCCATCTGT TGCTGCGCGG
          361 CCAGCCGGGA ACCGCTCAAC TCGGCCATGC GCGGGCCGAT CTCGGCGAAC ACCGCCCCCG
          421 CTTCGACGCT CTCCGGCGTG GTCCAGACCG CCACCGCGGC GCCGTCGTCC GCGACCCACA
          481 CCTTGCCGAT GTCGAGCCCG ACGCGCGTGA GGAAGAGTTC TTGCAGCTCG GTGACCCGCT
          541 CGATGTGGCG GTCCGGATCG ACGGTGTGGC GCGTGGCGGG GTAGTCGGCG AACGCGGCGG
          601 CGAGGGTGCG TACGGCCCTG GGGACGTCGT CGCGGGTGGC GAGGCGCACC GTGGGCTTGT
          661 ACTCGGTCAT GGTAAGCTCC GGTGGATCCC CCTGGGGAGA GAGGTCGGTG ATTCGGTCAA
          721 CGAGGGAGCC GACTGCCGAC GTGCGCTCCG GAGGCTTGCA GAATGCGGAA CACCGCGCGG
          781 GCAGGAACAG GGCCCACACT ACCGCCCCAC ACCCCGCCTC CCGCACCGCC CCTTCCCGGC
          841 CGCTGCTCTC GGCGCGCCCT GCTGAGCAGC CGCTATTGGC CACAGCCCAT CGCGGTCGGC
          901 GCGCTGCCAT TGCTCCCTGG CGCTGTCCGT CTGCGAGGGT ACTAGTGAGA CGTGCGGCTT
          961 CCGTTTGTCA CGTCCGGGCA CGCCGCGAAC CGCAAGGAAC CTTTCCCCGA CTTAGGGGCG
         1021 GAGCAAGAAA GCGTCGCCTG GGGGTCCCAA CAAGGGTAGC CGGCTAAGAT CCGGATGACG
         1081 CTGCCGAACC GAACGTGAAG AGTGTTGCCG AGACTCAGGA TCCGCGCCCG CTTGCCGTTT
        1141 TCTCCCCGG
        //
        Primer: M13R
        Region: insert 5' (AAVS1 arm1)
        Sequence file: RDB13918_A6B8h.seq check
        >D03070A02_A6B8_1_M13R_A10_01.ab1
            1 CATCTGATAC CCTCACTAAA GGGACTAGTC CTGCAGGTTT AAACGAATTC GCCCTTTGCT
           61 TTCTCTGACC AGCATTCTCT CCCCTGGGCC TGTGCCGCTT TCTGTCTGCA GCTTGTGGCC
          121 TGGGTCACCT CTACGGCTGG CCCAGATCCT TCCCTGCCGC CTCCTTCAGG TTCCGTCTTC
          181 CTCCACTCCC TCTTCCCCTT GCTCTCTGCT GTGTTGCTGC CCAAGGATGC TCTTTCCGGA
          241 GCACTTCCTT CTCGGCGCTG CACCACGTGA TGTCCTCTGA GCGGATCCTC CCCGTGTCTG
          301 GGTCCTCTCC GGGCATCTCT CCTCCCTCAC CCAACCCCAT GCCGTCTTCA CTCGCTGGGT
          361 TCCCTTTTCC TTCTCCTTCT GGGGCCTGTG CCATCTCTCG TTTCTTAGGA TGGCCTTCTC
          421 CGACGGATGT CTCCCTTGCG TCCCGCCTCC CCTTCTTGTA GGCCTGCATC ATCACCGTTT
          481 TTCTGGACAA CCCCAAAGTA CCCCGTCTCC CTGGCTTTAG CCACCTCTCC ATCCTCTTGC
          541 TTTCTTTGCC TGGACACCCC GTTCTCCTGT GGATTCGGGT CACCTCTCAC TCCTTTCATT
          601 TGGGCAGCTC CCCTACCCCC CTTACTCTCT AGTCTGTGCT AGCTCTTCCA GCCCCCTGTC
          661 ATGGCATCTT CCAAGGGTCC GAGAGCTCAG CTAGTCTTCT TCTCCACCGG GCCCCTATGT
          721 CACTCAGACA GCATGTTTGC TGCCTCAAGG GATCTTGTGT CCCTAGCTGG ACCACCTTTA
          781 ATTCCAGGGC GTAATGTGCT CTGATCTAGT ACTTATCTGT CCCTCCACCC CAATGGCATA
          841 TAAATCATTC CGGCTAATAT GTCTAGCATA TGGAATCCGT TACATAACTT ACAGGTAATG
        //
        Primer: M13_-40
        Region: insert 3' (AAVS1 arm2)
        Sequence file: RDB13918_A6B8i.seq check
        >D03070A02_A6B8_1_M13(-40)_B10_04.ab1
            1 CGCCGGCATG TATACGACTC ACTATAGGGC GAATTGAATT TAGCGGCCGC GAATTCGCCC
           61 TTAGAGCAGA GCCAGGAACC CCTGTAGGGA AGGGGCAGGA GAGCCAGGGG CATGAGATGG
          121 TGGACGAGGA AGGGGGACAG GGAAGCCTGA GCGCCTCTCC TGGGCTTGCC AAGGACTCAA
          181 ACCCAGAAGC CCAGAGCAGG GCCTTAGGGA AGCGGGACCC TGCTCTGGGC GGAGGAATAT
          241 GTCCCAGATA GCACTGGGGA CTCTTTAAGG AAAGAAGGAT GGAGAAAGAG AAAGGGAGTA
          301 GAGGCGGCCA CGACCTGGTG AACACCTAGG ACGCACCATT CTCACAAAGG GAGTTTTCCA
          361 CACGGACACC CCCCTCCTCA CCACAGCCCT GCCAGGACGG GGCTGGCTAC TGGCCTTATC
          421 TCACAGGTAA AACTGACGCA CGGAGGAACA ATATAAATTG GGGACTAGAA AGGTGAAGAG
          481 CCAAAGTTAG AACTCAGGAC CAACTTATTC TGATTTTGTT TTTCCAAACT GCTTCTCCTC
          541 TTGGGAAGTG TAAGGAAGCT GCAGCACCAG GATCAGTGAA ACGCACCAGA CAGCCGCGTC
          601 AGAGCAGCTC AGGTTCTGGG AGAGGGTAGC GCAGGGTGGC CACTGAGAAC CGGGCAGGTC
          661 ACGCATCCCC CCCTTCCCTC CCACCCCCTG CCAAGCTCTC CCTCCCAGGA TCCTCTCTGG
          721 CTCCATCGTA AGCAAACCTT AGAGGTTCTG GCAAGGAGAG AGATGGCTCC AGGAAATGGG
          781 GGGTGTGTCA CCAGATAAGG AATCTGCCTA ACAGGAGGTG GGGGTTAGAC CCAATATCAG
          841 AGACTAGGAA AGGAGGAGGC CTAAAGATGG GGCTTTTCTG TCACCAATCC TGTCCCTAGT
          901 AACCAATAGA GCCCACCGCA TCCCCAGCAT GCCTGCTATT GTCTTCCCAA TCCTCCCCCT
          961 TGCTGTCCTG CCCCCACCCC AACCCCCCAG ATAGATGACA CTACTCAGAA CAATGCGATG
         1021 CAATTCCTTC ATTTTATAGA AGACAGTGGG GAGTGCACGT CATGGTCAAG GAAGCACGGG
        1081 AGGGTCTACA ACGAATGCTG GCAACCTTAG GAAAAG
        //
        Primer: SV40polyA-R
        Region: insert mid (OsTIR1 3')
        Sequence file: RDB13918_A6B8j.seq check
        >D03070A02_A6B8_1_SV40polyA-R_C10_07.ab1
            1 GGTTTAGCAG TAAACCTCTA CAAATGTGGT ATGGCTGATT ATGATCTAGA GTCGCGGCCC
           61 CGGTGGATCC CGGGCCCGCG GTACCGTCGA CGGGCCCGGG ATCCGATGGA TCCTCACAGA
          121 ATCTTCACAA AGTTGGGAGC ATCATCTCTT GCCCCTGCGG TGGTCCGATA CACATACAGT
          181 TTCTCCACCT TTGGCAGGTC GCCATGATTT TCCTCCATTT CGTTAGACCC GTCCCGCTCA
          241 TTAATGACTT CCACGTTCAG CATAGGCATT TTACTAGCCA GGACCTGACA GCCCTTCAGA
          301 GTCACATTGC AACTAGACAT CCACAGGCTC CTCATTGTCT CGTATCTGGC AAAGTTTCCC
          361 AGCAGAGCGG CATCCCCGAA AGGGCTGTCC CGGATTTCCA GTTTTCGCAG ATTCTTACAC
          421 CCGTTCATCA CGTGCATCAT GCCCTTATCG GAGTCTCCGG CAAATGCAAT GCTCAGCATC
          481 TCCAGCTGCT TAGCATACTT GCCGATGTAC ATGAACACTT TGTCTGTCAG CAGGCCGGAA
          541 ATGCTCAGCC TCTGGAGTCC CTTGCACTCT CTCACGATAG CTCCAAACCC TTCATCCAGT
          601 GGCTGGGATG TGACCACGTC GGGTTTGCCT GGTTCCAGAA TACACAGCCG AAATCGGGTG
          661 AAAATTAGGG CAGTTCTTGG CGACGGTCAC CAGTGCAGCA TTAGTCATCT GATGACAGAA
        721 ATACAGCAGG CTGTTCAGCT TTGGGCATCC CAGTGAAGAC TGCCA
        //

        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 Sathyan, K.M., An improved auxin-inducible degron system preserves native protein levels and enables rapid and specific protein depletion. Genes Dev. 33 (19-20): 1441-1455 (2019). PMID 31467088.
        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.

        2022.05.17

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