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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 for distribution 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。
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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.

        How to cite this biological resource

        Materials & Methods section:

        The pMK232 (AAVS1 CMV-OsTIR1-PURO) was provided by the RIKEN BRC through the National BioResource Project of the MEXT, Japan (cat. RDB13918).

        Reference section:

        Natsume, T., Kiyomitsu, T., Saga, Y., Kanemaki, M.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. [link to RRC of NBRP]

        Further references such as user reports and related articles (go to bottom)


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        Featured content

        Featured content Auxin Inducible Degron (AID) System (English text)
        Featured content Auxin Inducible Degron (AID) System (Japanese text)

        QC test results

        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_A6B8p1-4.pdf check

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

        Primer: M13R (Pr0160)
        Region: insert mid(AAVS1 arm1 mid)
        Sequence file: RDB13918_A6B8o.seq check
        >D03070A02_A6B8_1_M13R_A10_01.ab1
            1 NNNNNNNNNN CNNNNNNAAN GGGNNTAGTC CTGCAGGTTT AAACGAATTC GCCCTTTGCT
           61 TTCTCTGNCN 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 TTTCTTANGA TGGCCTTCTC
          421 CGACNGANGT CTCCCTTGCG TCCCGCCTCC CCTTCNTGTN NNNCTGCATC ATCACCGTTT
          481 TTCTGGACAA NCCCNAAGTA CCCCGTCTNC CNGGCTTTAN CCACCTCTNC ATNNNCTTGN
          541 TTTCTTNGCC NNNNNNCCCC NNNCTNNNGN GNANTNNNNT CANNNNNNAN TNNNTTNNNT
        601 NNGGNANCTN CCCNANCCCC NNNNNNNNNN
        //
        Primer: M13_-40 (Pr0410)
        Region: T7 pro,insert 3'(AAVS1 arm2 3')
        Sequence file: RDB13918_A6B8p.seq check
        >D03070A02_A6B8_1_M13(-40)_B10_04.ab1
            1 NNNNNNNNNN NANNNGNNTN NNNNTAGGGC 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 TTTCCNAACT GCTTCTCCTC
          541 TTGGGAAGTG TAAGGAAGCT GCAGCACCAG GATCAGTGAA ACGCACCAGA CAGCCGCGTC
          601 AGANCAGCTC ANGTTCTGGG AGAGGGTAGC GCANGGTGGC CACTGAGAAC CGGGCANGTC
          661 ACGCATCCCC CCCTTCCCTC CCACCCCCTG CCNAGCTCTC CCTCCCANGN TCCTCTCTGG
          721 CTCCNTCGTA AGCAAACCTT ANAGGNTCTG GCAAGGAGAG AGANGGNTCC AGGAAATGGG
          781 GGGTGTGTCA CCAGATANNN AATCTGCCNN ACNNNANNTG GGGGTTAGAN CCNANATCNN
          841 NNANTAGNNA NGNNNNAGNN CNNANNNNNG GGCTTTNNNG TCNNNNNTNC TGNNCCTANN
        901 NANCNNNANA NCCNANNGNN NNCCNNNNNN NNNN
        //
        Primer: CMV_Forward (Pr0016)
        Region: insert mid(CMV pro,OsTIR1 5')
        Sequence file: RDB13918_A6B8q.seq check
        >D03070A02_A6B8_1_CMV_Forward_D10_10.ab1
            1 NNNNNNNNNN NNNNNTNNNN GCNNTGGGCG GTANGCGTGT 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 GACTTTNACC TGGTGCCCCC TGATTGGGGA
          361 GGANACNCCN GNCCTTGGAT CGAGGNAGCC GCTCGCGGCT GTCNNGGACT GGANGAACTG
          421 NNCNTGAANC GAANGGNGGN CTCTGANNAA AGTCNGGNNC TGCNNNNNNN NANCNNNCNN
          481 NNGNTNNNNN NNNNGNNNNT GANTTNNNNN NNNANNNNNN CNNNNCNNNN NGNNNNNNNN
        541 NN
        //
        Primer: SV40polyA_R2 (Pr0623)
        Region: insert mid(OsTIR1 3')
        Sequence file: RDB13918_A6B8r.seq check
        >13918_13918_A9KrF_SV40polyA_R2_D09_12_ABI24.ab1
            1 NNNNNNNNNN NGNANNNNNN GTTAANNANN ACNATTGCAT TCNTTTTATG TTTCNGGTTC
           61 AGGGGGAGGT GTGGGAGGTT TTTTAAAGCA AGTAAAACCT CTACAAATGT GGTATGGCTG
          121 ATTATGATCT AGAGTCGCGG CCCCGGTGGA TCCCGGGCCC GCGGTACCGT CGACGGGCCC
          181 GGGATCCGAT GGATCCTCAC AGAATCTTCA CAAAGTTGGG AGCATCATCT CTTGCCCCTG
          241 CGGTGGTCCG ATACACATAC AGTTTCTCCA CCTTTGGCAG GTCGCCATGA TTTTCCTCCA
          301 TTTCGTTAGA CCCGTCCCGC TCATTAATGA CTTCCACGTT CAGCATAGGC ATTTTACTAG
          361 CCAGGACCTG ACAGCCCTTC AGAGTCACAT TGCAACTAGA CATCCACAGG CTCCTCATTG
          421 TCTCGTATCT GGCAAAGTTT CCCAGCAGAG CGGCATCCCC GAAAGGGCTG TCCCGGATTT
          481 CCAGTTTTCG CAGATTCTTA CACCCGTTCA TCACGTGCAT CATGCCCTTA TCGGAGTCTC
          541 CGGCAAATGC AATGCTCAGC ATCTCCAGCT GCTTAGCATA CTTGCCGATG TACATGAACA
          601 CTTTGTCTGT CAGCAGGCCG GAAATGCTCA GCCTCTGGAG TCCCTTGCAC TCTCTCACGA
          661 TAGCTCCAAA CCCTTCATCC AGTGGCTGGG ANGTGACCAC GTCGGGNTTG CCTGGTTCCA
          721 GAATACACAG CCGAANTCGG GTGAAATTAG GGCAGTTCTT GGCGACNGTC ACCAGTGCAG
          781 CATTAGTCAT CTGATGACAG AAATACAGCA NGNTGTTCNN CTTTGGGCAT CCCAGTGAGA
          841 NTGNCACCAN GNNNTNCTCA NNGACNGCAC TATATNCNNN CNCNTAAAAA TCNNANNGGN
          901 ANNNTCNCAN CTNCNNGANG NNCTNNNNNN ANCTNNNNAN NNCCNNGNNC CNNNNNNNNN
        961 N
        //
        Primer: BGH_rev2 (Pr0606)
        Region: insert mid(bGH pA,PuroR,hPGK pro)
        Sequence file: RDB13918_A6B8s.seq check
        >D03070A02_A6B8_1_BGH_rev2_E10_13.ab1
            1 NNNNNNNNNN NNNNNNNNNN GGGGGNNGGG CNNACNACAG 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 GANGCTTGCA GANTGCGGAN NACCGCGCGG
          781 GCANGAACAG GGCCCACACT ACCGCNNCNC ACCCCGCCTC CCGCACNGCC CCTTCCCGGC
          841 CGCTGCTCTC GGCGCGNCCT GCTGANCAGC CGCTNTTGGC CNCAGCCNNT CNNNGNCGGC
          901 GCGCTGCNNT NNCTNCCTNN CGCTGNNCGT CTGCGAGGGN NNNNNNNAGA CGTGNNNCTT
          961 NCGNTTNNNN CNNNNNGGCA NGCCNCNNNN NGNNNGNNNN NNTNNCCNNN NNTNNNNNNN
        1021 NNN
        //

        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. [link to RRC of NBRP]
        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.

        2023.10.05

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