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pcDNA3.1 FLAG-hRFWD3 I639K (#RDB17492)

Expression vector of human ring finger and WD repeat domain 3 (RFWD3) mutant, I639K, chromatin localization-defective mutation.

Clone info. Expression vector of human ring finger and WD repeat domain 3 (RFWD3) mutant, I639K, chromatin localization-defective mutation, tagged with FLAG at N-terminus.
Comment Expression was confirmed by the depositor with immunoblotting. Known differences (at least): G to A substitution at 1269nt of NM_018124.4 (R392K; AGG to AAG).
Vector backbone pcDNA3.1/Zeo (plasmid)
Selectable markers E. coli, Ampicillin; mammalian cells, Zeocin.
Gene/insert name Human RFWD3 cDNA
Depositor|Developer Takata, Minoru |
 

Distribution information

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Terms and conditions for distribution In publishing the research results obtained by use of the BIOLOGICAL RESOURCE, a citation of literature designated by the DEPOSITOR is requested. (Inano, S et al. Mol. Cell, 66(5): 622-634, 2017)
Remarks
Gateway® Expression clone. Please refer Life Technologies Corporation signs license agreement with RIKEN BRC.
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MTAに書く使用条件 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 (Inano, S et al. Mol. Cell, 66(5): 622-634, 2017)。
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Catalog # Resource name Shipping form Fee (non-profit org.)
RDB17492 pcDNA3.1 FLAG-hRFWD3 I639K DNA solution JPY 9,460 (not-for-profit academic purpose)
plus cost of shipping containers, dry ice (if required) and shipping charge plus licensing fee equivalent for Gateway® expression clone

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 pcDNA3.1 FLAG-hRFWD3 I639K was provided by the RIKEN BRC through the National BioResource Project of the MEXT, Japan (cat. RDB17492).

Reference section:

Inano, S., Sato, K., Katsuki, Y., Kobayashi, W., Tanaka, H., Nakajima, K., Nakada, S., Miyoshi, H., Knies, K., Takaori-Kondo, A., Schindler, D., Ishiai, M., Kurumizaka, H., Takata, M., RFWD3-Mediated Ubiquitination Promotes Timely Removal of Both RPA and RAD51 from DNA Damage Sites to Facilitate Homologous Recombination. Mol. Cell 66 (5): 622-634.e8 (2017). PMID 28575658. [link to RRC of NBRP]

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

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 RDB17492_A9Iop1-1.pdf check

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

Primer: CMV_Forward (Pr0016)
Region: CMV pro, FLAG, attB1, insert 5'
Sequence file: RDB17492_A9Ioa.seq check
>D06551A3_A9Io_2_CMV_Forward_G01_19_ABI24.ab1
    1 NNNNNNNNNN NNNNNNNNNN NNNNNGNNGC AATGGGCGGT AGGCGTGTAC GGTGGGAGGT
   61 CTATATAAGC AGAGCTCTCT GGCTAACTAG AGAACCCACT GCTTACTGGC TTATCGAAAT
  121 TAATACGACT CACTATAGGG AGACCCAAGC TGGCTAGCAC CATGGATTAT AAAGATGATG
  181 ACGATAAGCA TGGTGGTTCT GGTCCAAGCT ATCAAACAAG TTTGTACAAA AAAGCAGGCT
  241 CCGCGGCCGC CCCCTTCACC ATGGCTCATG AAGCAATGGA ATATGATGTT CAGGTGCAGT
  301 TAAATCATGC CGAACAACAG CCAGCTCCTG CTGGCATGGC CAGCAGCCAA GGGGGACCAG
  361 CCCTCCTCCA GCCTGTTCCT GCTGATGTGG TCAGCAGCCA GGGGGTACCA TCCATCCTCC
  421 AGCCAGCTCC TGCTGAGGTG ATCAGCAGCC AAGCGACACC ACCCCTGCTC CAGCCTGCTC
  481 CGCAACTGTC TGTTGACCTG ACAGAAGTGG AGGTCTTGGG AGAAGACACT GTGGAGAACA
  541 TCAATCCAAG AACTTCAGAA CAACATAGGC AGGGATCTGA TGGTAATCAC ACCATCCCAG
  601 CATCTTCGTT GCATTCAATG ACCAACTTCA TCAGCGGACT GCAGAGACTT CATGGCATGC
  661 TGGAATTCCT GAGACCTTCA TCTTCAAACC ACAGTGTAGG GCCAATGAGA ACAAGAAGGA
  721 NGGTATCTGC TTCACGGAGG GCAAGAGCCG GAGGGTCTCA GANGACAGAC AGTGCCAGGT
  781 TGAGAGCACC ATTGGATGCT TACTTTCANG TGAGCAGGAN CCAGCCTGAC TTGCCAGCTA
  841 CCACTTATGA TTCAGAGANT AGGAATCCTG TATCTGAAGA NTTGCAGGTG TCTAGTAGTT
  901 CTGATTCTGA NAGTGACAGC TCTGCANANT ATNNAGGGGT TGTTGNNNNN NNANANGAAT
  961 CTGGANCTGN CATTTTANAA GANCNNCNNG CANNNGNCNC ANCNNANCNN GNANNTTACA
1021 TNNNTCNNNG NNNNNNNNNN
//
Primer: f1ori_R3 (PR0142)
Region: bGHp(A), attB2, insert 3' (I639K)
Sequence file: RDB17492_A9Iob.seq check
>D06551A3_A9Io_2_f1ori_R3_C03_09_ABI24.ab1
    1 NNNNNNNNNN NNNNNNNNNN NNNGNNCTTT CNGCCTNNGA AGCCNTAGAG CCCACCGCAT
   61 CCCCAGCATG CCTGCTATTG TCTTCCCAAT CCTCCCCCTT GCTGTCCTGC CCCACCCCAC
  121 CCCCCAGAAT AGAATGACAC CTACTCAGAC AATGCGATGC AATTTCCTCA TTTTATTAGG
  181 AAAGGACAGT GGGAGTGGCA CCTTCCAGGG TCAAGGAAGG CACGGGGGAG GGGCAAACAA
  241 CAGATGGCTG GCAACTAGAA GGCACAGTCG AGGCTGATCA GCGGGTTTAT CGAACCACTT
  301 TGTACAAGAA AGCTGGGTCG GCGCGCCCAC CCTTTCACTC CCACTTATAG ATGTGGACCA
  361 TCTTCTCTGT TAAGGTAGCC AAGTAGCTGT TACGGTTCAC CTCAAATGGG CAGATGTCCA
  421 ACACAGGCTG ATCGGTCTGT AGGTCCTGGA GCAACGAGCC ACTGGCAGCA TCCCACAGCA
  481 GGGCAGAATT TGCTGCTTCA TCCCCAGTAC ACACCAGGAT GTTGCCATCA TTCTCTGGGC
  541 TTTGGAAAAT GGCATTTTTG GTCAATAGTT TGCAAGTAGG TCCTCCAAAA AATGTATGTA
  601 CAGGCTGGCA GGAGCAGATT GGATTTCCAG TGTCATCCAG TCGGTAGGAC ATTTCCATCA
  661 GCACACTTCG TATGGTGGTG TGATTTTTAT CAGGCCTGTA GGTCACAAGA CAGTGCCGGG
  721 AGCTGTTCTC TGTCTGAAAG TCTTTGCACC CCCCTGGCTC CAAGGGCAGC ACATGAGGCC
  781 AATGAGAAAA GTCCATTTTC TGTTCCCAGA ATGAAGCATC CTCCAAGGTT CCAGCCAGCA
  841 CCCCACCATA TGGAAATGCA GCTGANGCAG CTCTGGGCAT GTATGACAGG GANACCAGNG
  901 GGCATCTGNC TTTCTGANCT ACTAACTCCT GCNCATGACT GCNCNNGTTT CNCANGNCAT
  961 ATNCCAGANN TGANNNATNG GNNAGTCCAN CATANNTGTA NTNAGNCNCA TNNNNNNNNC
1021 NNCNNCNNNN NNNNNN
//
Primer: hRFWD3-N_F (Pr0753)
Region: insert mid
Sequence file: RDB17492_A9Ioc.seq check
>D06551A3_A9Io_2_hRFWD3-N_F_B04_04_ABI24.ab1
    1 NNNNNNNNNN NNNNNNNNAT CTGANANTTG CAGGTGTCTA GTAGTTCTGA TTCTGACAGT
   61 GACAGCTCTG CAGAGTATGG AGGGGTTGTT GACCAGGCAG AGGAATCTGG AGCTGTCATT
  121 TTAGAAGAGC AACTAGCAGG TGTCTCAGCA GAGCAAGAAG TTACATGTAT CGATGGAGGC
  181 AAGACCCTCC CCAAACAGCC ATCTCCCCAG AAGTCTGAGC CTCTGCTACC TTCTGCTTCT
  241 ATGGATGAGG AAGAAGGGGA CACTTGTACA ATATGTCTGG AACAGTGGAC CAATGCTGGG
  301 GACCACCGGC TCTCAGCATT ACGCTGTGGG CATCTCTTTG GGTATAGGTG CATTTCCACG
  361 TGGCTTAAAG GACAAGTACG AAAATGTCCC CAGTGCAACA AGAAAGCCAG GCACAGTGAC
  421 ATTGTCGTCC TTTATGCCCG AACCCTGAGA GCTTTGGACA CTAGTGAACA GGAGCGCATG
  481 AAAAGTTCCC TACTGAAGGA ACAGATGCTA AGGAAACAGG CCGAGTTAGA ATCAGCACAG
  541 TGCCGACTCC AACTGCAGGT CCTCACTGAT AAGTGCACTA AGCTTCAAAG GCGTGTTCAG
  601 GACTTGCAAA AACTTACGTC ACATCAAAGT CAGAATTTAC AGCAACCCAG GGGCTCCCAA
  661 GCATGGGTCC TGAGCTGCTC ACCCTCCAGC CAGGGCCAGC ACAAGCACAA GTACCACTTC
  721 CAAAAGANCT TCACAGTATC TCAGGCAGGA AACTGCCGGA TCATGGCATA CTGTGATGCT
  781 CTGANCTGCC TGGTGATATC ACAGCCTTCT CCTCANGCCT CTTTTCTTCC AGGCTTTGGT
  841 GTTAAGANGT TGAGTACTGC CAACATGAAG AGCAGTCAGT ACATTCCGAT GCNTGGCAAA
  901 CAGATCCGNN NANNGGCGTT TANCAGTTAN CTCANANGNT TGCTACTCTC TGCTTCCCTA
961 NANNACACTA TTAAACNGAN NNNCCNNNNN NNNAANANNN NNNNNNN
//
Primer: SV40pro_ori_F (Pr0731)
Region: SV40pro_ori, bleoR, SV40p(A)
Sequence file: RDB17492_A9Iod.seq check
>D06551A3_A9Io_2_SV40pro_ori_F_A02_02_ABI24.ab1
    1 NNNNNNNNNN NNNNNNNNNN NCGCCTCTGC CTCTGANCTA TTCNNNAAGT AGTGAGGAGG
   61 CTTTTTTGGA GGCCTAGGCT TTTGCAAAAA GCTCCCGGGA GCTTGTATAT CCATTTTCGG
  121 ATCTGATCAG CACGTGTTGA CAATTAATCA TCGGCATAGT ATATCGGCAT AGTATAATAC
  181 GACAAGGTGA GGAACTAAAC CATGGCCAAG TTGACCAGTG CCGTTCCGGT GCTCACCGCG
  241 CGCGACGTCG CCGGAGCGGT CGAGTTCTGG ACCGACCGGC TCGGGTTCTC CCGGGACTTC
  301 GTGGAGGACG ACTTCGCCGG TGTGGTCCGG GACGACGTGA CCCTGTTCAT CAGCGCGGTC
  361 CAGGACCAGG TGGTGCCGGA CAACACCCTG GCCTGGGTGT GGGTGCGCGG CCTGGACGAG
  421 CTGTACGCCG AGTGGTCGGA GGTCGTGTCC ACGAACTTCC GGGACGCCTC CGGGCCGGCC
  481 ATGACCGAGA TCGGCGAGCA GCCGTGGGGG CGGGAGTTCG CCCTGCGCGA CCCGGCCGGC
  541 AACTGCGTGC ACTTCGTGGC CGAGGAGCAG GACTGACACG TGCTACGAGA TTTCGATTCC
  601 ACCGCCGCCT TCTATGAAAG GTTGGGCTTC GGAATCGTTT TCCGGGACGC CGGCTGGATG
  661 ATCCTCCAGC GCGGGGATCT CATGCTGGAG TTCTTCGCCC ACCCCAACTT GTTTATTGCA
  721 GCTTATAATG GTTACAAATA AAGCAATAGC ATCACAAATT TCACAAATAA AGCATTTTTT
  781 TCACTGCATT CTAGTTGTGG TTTGTCCAAA CTCATCAATG TATCTTATCA TGTCTGTATA
  841 CCGTCGANCT CTANCTAGAG CTTGGCGTAA TCATGGTCAT ANCTGTTTNC TGTGTGAAAT
  901 TGTNATCCGC TCACNATTNN NCACAACATA CGAGCCNGNA AGCATAAAGT GNAANNCNGG
  961 GGGNGNCNNA TGAGTGANCT AACNCACATT ANTTGNNTTG CGCNCNNNGN CNNCTTTNCA
1021 GNNNGNNANN NNNNNNNN
//

Please visit Sequencing and PCR primers for primer information.

References

Original, user report and related articles

original Inano, S., RFWD3-Mediated Ubiquitination Promotes Timely Removal of Both RPA and RAD51 from DNA Damage Sites to Facilitate Homologous Recombination. Mol. Cell 66 (5): 622-634.e8 (2017). PMID 28575658. [link to RRC of NBRP]