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

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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RDB13919 pMK243 (TetOne OsTIR1 AAVS1) 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 RDB13919_A6B8p1_v2.pdf

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

Primer: TetOne Oligo 1 RDB13919_A6B8a_2.seq
>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 RDB13919_A6B8b_2.seq
>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 RDB13919_A6B8c_2.seq
>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 RDB13919_A6B8d_2.seq
>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 RDB13919_A6B8e_2.seq
>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 RDB13919_A6B8f_2.seq
>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

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 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