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pcDNA3.1 FLAG-hRFWD3 LQP/AAA (#RDB17495)

Expression vector of human ring finger and WD repeat domain 3 (RFWD3) mutant, LQP/AAA, LQP motifs replaced by triple alanines.

Clone info. Expression vector of human ring finger and WD repeat domain 3 (RFWD3) mutant, LQP/AAA, LQP motifs replaced by triple alanines, tagged with FLAG at N-terminus, see Fig. S5B of Inano, S. et al., Mol. Cell, 66 (5): 622-634, 2017.
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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MTA, for use for not-for-profit academic purpose [Word] [Example of MTA ]
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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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提供依頼書  [依頼書の記入例 ]
提供同意書 (MTA, 非営利学術目的用)[Word] [MTAの記入例 ]
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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)。
備考
Gateway®テクノロジーを用いた発現クローンです。「Gateway®テクノロジーを用いたクローンの提供と寄託のご案内」をご覧ください。

Catalog # Resource name Shipping form Fee (non-profit org.)
RDB17495 pcDNA3.1 FLAG-hRFWD3 LQP/AAA 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 LQP/AAA was provided by the RIKEN BRC through the National BioResource Project of the MEXT, Japan (cat. RDB17495).

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 RDB17495_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' (LQP/AAA)
Sequence file: RDB17495_A9Ioa.seq check
>D06551A6_A9Io_1_CMV_Forward_E02_14_ABI24.ab1
    1 NNNNNNNNNN NNNNNNNNNN NNNNNNNGCN NNTGGGCGGT 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 CGAAGCAGCT GCAGCTCCTG CTGGCATGGC CAGCAGCCAA GGGGGACCAG
  361 CCCTCGCTGC AGCTGTTCCT GCTGATGTGG TCAGCAGCCA GGGGGTACCA TCCATCGCAG
  421 CTGCAGCTCC TGCTGAGGTG ATCAGCAGCC AAGCGACACC ACCCCTGGCA GCAGCTGCTC
  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 GGGTATCTGC TTCACGGAGG GCAAGANCCG GAGGGTCTCA GANGACANAC AGTGCCAGGT
  781 TGAGAGCACC ATTGGATGCT TACTTTCNGG TGAGCAGGAN CCAGCCTGAC TTGCCNGCTA
  841 CCACTTATGA TTNNNANACT AGGAATCCTG TATCTGAAGA GTTGCNGGTG TCTAGTANTT
  901 CTGATTCTGA NNGTGACAGC TCTGCANANT ATGGANGGGT TGTTGNNNNN GNNNNAGGAA
  961 TCNNGANCTG NCNTTTTANA AGANNNACTN GNNNNGTGNN TCANNNNNNC NANNAANTTA
1021 CNNNNNNNNN TGGNNNNNNN NNN
//
Primer: BGH_rev2 (Pr0606)
Region: bGH p(A), attB2, insert 3'
Sequence file: RDB17495_A9Iob.seq check
>D06551A6_A9Io_1_BGH_rev2_A03_03_ABI24.ab1
    1 NNNNNNNNNN NNNNNGNNNG GGNGNNGGGC AAACNACAGA TGGCTGGCAA CTAGAAGGCA
   61 CAGTCGAGGC TGATCAGCGG GTTTATCGAA CCACTTTGTA CAAGAAAGCT GGGTCGGCGC
  121 GCCCACCCTT TCACTCCCAC TTATAGATGT GGACCATCTT CTCTGTTAAG GTAGCCAAGT
  181 AGCTGTTACG GTTCACCTCA AATGGGCAGA TGTCCAACAC AGGCTGATCG GTCTGTAGGT
  241 CCTGGAGCAA CGAGCCACTG GCAGCATCCC ACAGCAGGGC AGAATTTGCT GCTTCATCCC
  301 CAGTACACAC CAGGATGTTG CCATCATTCT CTGGGCTTTG GAAAATGGCA TTTTTGGTCA
  361 ATAGTTTGCA AGTAGGTCCT CCAAAAAATG TATGTACAGG CTGGCAGGAG CAGATTGGAT
  421 TTCCAGTGTC ATCCAGTCGG TAGGACATTT CCATCAGCAC ACTTCGTATG GTGGTGTGAT
  481 TTTTATCAGG CCTGTAGGTC ACAAGACAGT GCCGGGAGCT GTTCTCTGTC TGAAAGTCTA
  541 TGCAGCCCCC TGGCTCCAAG GGCAGCACAT GAGGCCAATG AGAAAAGTCC ATTTTCTGTT
  601 CCCAGAATGA AGCATCCTCC AAGGTTCCAG CCAGCACCCC ACCATATGGA AATGCAGCTG
  661 AGGCAGCTCT GGGCATGTAT GACAGGGAGA CCAGTGGGCA TCTGGCTTTC TGAGCTACTA
  721 ACTCCTGCAC ATGACTGCTC GTGTTTCGCA CGTCATATAC CAGAATTGAA CCATTGGCCA
  781 GTCCAGCATA GATGTAGTTA GCCTCATCAA GACACCAGCA ACAGCTCCNG ANAGGACGTC
  841 CAGCATTATA AGTCTGGANC ACGGNATTTG TCTCCAGGCT GGTCANTTTA ATAGNGTTGT
  901 CNANGGAANC ANANANTANC AAGCCTCTGA NGTAANTGCT AANNGCCANT NNNNNGNATC
  961 NGNTTNNCNT GCNTCNGANN GNNCTGACTG NNNNNCNTGN TNGNNAGTAN NNNNNATNNN
1021 NNNNNNN
//
Primer: hRFWD3-N_F (Pr0753)
Region: insert mid
Sequence file: RDB17495_A9Ioc.seq check
>D06551A6_A9Io_1_hRFWD3-N_F_B01_04_ABI24.ab1
    1 NNNNNNNNNN NNNNNNNNTC TGANANTTGC AGGTGTCTAG TAGTTCTGAT TCTGACAGTG
   61 ACAGCTCTGC AGAGTATGGA GGGGTTGTTG ACCAGGCAGA GGAATCTGGA GCTGTCATTT
  121 TAGAAGAGCA ACTAGCAGGT GTCTCAGCAG AGCAAGAAGT TACATGTATC GATGGAGGCA
  181 AGACCCTCCC CAAACAGCCA TCTCCCCAGA AGTCTGAGCC TCTGCTACCT TCTGCTTCTA
  241 TGGATGAGGA AGAAGGGGAC ACTTGTACAA TATGTCTGGA ACAGTGGACC AATGCTGGGG
  301 ACCACCGGCT CTCAGCATTA CGCTGTGGGC ATCTCTTTGG GTATAGGTGC ATTTCCACGT
  361 GGCTTAAAGG ACAAGTACGA AAATGTCCCC AGTGCAACAA GAAAGCCAGG CACAGTGACA
  421 TTGTCGTCCT TTATGCCCGA ACCCTGAGAG CTTTGGACAC TAGTGAACAG GAGCGCATGA
  481 AAAGTTCCCT ACTGAAGGAA CAGATGCTAA GGAAACAGGC CGAGTTAGAA TCAGCACAGT
  541 GCCGACTCCA ACTGCAGGTC CTCACTGATA AGTGCACTAA GCTTCAAAGG CGTGTTCAGG
  601 ACTTGCAAAA ACTTACGTCA CATCAAAGTC AGAATTTACA GCAACCCAGG GGCTCCCAAG
  661 CATGGGTCCT GAGCTGCTCA CCCTCCAGCC AGGGCCAGCA CAAGCACAAG TACCACTTCC
  721 AAAAGANCTT CACAGTATCT CAGGCAGGAA ACTGCCGGAT CATGGCATAC TGTGNTGCTC
  781 TGAGCTGCCT GGTGATATCA CAGCCTTCTC CTCANGCCTC TTTTCTTCCN GGCTTTNNTG
  841 TTAAGANGTT GAGTACTGCN NACATGAAGA NCANNCAGTA CATTCNGATG NATGGCAAAC
  901 AGATNCGNGG NNNGGCGTTT ANCANTTACN TCAGANGCTT GCNACTNTCN GCTTCCNNAN
961 NNNACACTAT NAANNNGANN NGNCNNNNNN NNNANTANNN NNNNNNN
//
Primer: SV40pro_ori_F (Pr0731)
Region: SV40 pro_ori, bleoR
Sequence file: RDB17495_A9Iod.seq check
>D06551A6_A9Io_1_SV40pro_ori_F_B03_06_ABI24.ab1
    1 NNNNNNNNNN NNNNNNNNNN NNNCNCTGCC TCTGAGCNNT TCNNGAAGTA GTGAGGAGGC
   61 TTTTTTGGAG GCCTAGGCTT TTGCAAAAAG CTCCCGGGAG CTTGTATATC CATTTTCGGA
  121 TCTGATCAGC ACGTGTTGAC AATTAATCAT CGGCATAGTA TATCGGCATA GTATAATACG
  181 ACAAGGTGAG GAACTAAACC ATGGCCAAGT TGACCAGTGC CGTTCCGGTG CTCACCGCGC
  241 GCGACGTCGC CGGAGCGGTC GAGTTCTGGA CCGACCGGCT CGGGTTCTCC CGGGACTTCG
  301 TGGAGGACGA CTTCGCCGGT GTGGTCCGGG ACGACGTGAC CCTGTTCATC AGCGCGGTCC
  361 AGGACCAGGT GGTGCCGGAC AACACCCTGG CCTGGGTGTG GGTGCGCGGC CTGGACGAGC
  421 TGTACGCCGA GTGGTCGGAG GTCGTGTCCA CGAACTTCCG GGACGCCTCC GGGCCGGCCA
  481 TGACCGAGAT CGGCGAGCAG CCGTGGGGGC GGGAGTTCGC CCTGCGCGAC CCGGCCGGCA
  541 ACTGCGTGCA CTTCGTGGCC GAGGAGCAGG ACTGACACGT GCTACGAGAT TTCNATTCCN
  601 CCGCCGCCTT CTATGAAAGG NTGGGCTTCG NANTCNTTTT CCGGGACGCC GGCTGNATGA
  661 TCCTCCNGCG CNGGGATCTC ATGCTGGANT NCTNCGCCCN CCCNNACTTG NTTATTGNAG
  721 CTTATAATGG TTANAANTNA AAGCAATAGC ATCNCAAATT TCNNNANNAA AGCNNTTTTT
  781 TNNCTGNNTN CNNNTTGTGG NTTGNNNNAN CTCNTCNNNG NANNNNNNNC NNGNNNNGNN
841 NNNNCGNCNA CNTCNNNNTN NNNNNNNNN
//

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]