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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.
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 Kanemaki, Masato |
 
Other clones in our bank human AAVS1 (NCBI Gene 17) | Oryza sativa |

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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RDB13918 pMK232 (AAVS1 CMV-OsTIR1-PURO) DNA solution

cDNA Request [in Japanese] [in English]

References and tips

Electronic file

Featured content

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

References

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.

User reports and related articles (go to bottom)


Sequence information

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 RDB13918_A6B8p1_v2.pdf

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

Primer: CMV-Forward RDB13918_A6B8a_2.seq
>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 RDB13918_A6B8b_2.seq
>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 RDB13918_A6B8c_2.seq
>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 RDB13918_A6B8d_2.seq
>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 RDB13918_A6B8e_2.seq
>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

References

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 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 .

2018.12.11

GNP_filter3_RDBDEP_html_181012.pl