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pcDNA3.1 FLAG-hRFWD3 S46/63A (#RDB17493)

Expression vector of human ring finger and WD repeat domain 3 (RFWD3) mutant, S46/63A, non-phosphorylatable mutation.

Clone info. Expression vector of human ring finger and WD repeat domain 3 (RFWD3) mutant, S46/63A, non-phosphorylatable 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)。
備考
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Catalog # Resource name Shipping form Fee (non-profit org.)
RDB17493 pcDNA3.1 FLAG-hRFWD3 S46/63A 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 S46/63A was provided by the RIKEN BRC through the National BioResource Project of the MEXT, Japan (cat. RDB17493).

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 RDB17493_A9Iop1-1.pdf check

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

Primer: CMV_Forward (Pr0016)
Region: CMV pro, attB1, FLAG, insert 5' (S46A, S63A)
Sequence file: RDB17493_A9Ioa.seq check
>D06551A4_A9Io_1_CMV_Forward_B02_05_ABI24.ab1
    1 NNNNNNNNNN NNNNNNNNTG NNGCNNATGG GCGGTAGGCG TGTACGGTGG GAGGTCTATA
   61 TAAGCAGAGC TCTCTGGCTA ACTAGAGAAC CCACTGCTTA CTGGCTTATC GAAATTAATA
  121 CGACTCACTA TAGGGAGACC CAAGCTGGCT AGCACCATGG ATTATAAAGA TGATGACGAT
  181 AAGCATGGTG GTTCTGGTCC AAGCTATCAA ACAAGTTTGT ACAAAAAAGC AGGCTCCGCG
  241 GCCGCCCCCT TCACCATGGC TCATGAAGCA ATGGAATATG ATGTTCAGGT GCAGTTAAAT
  301 CATGCCGAAC AACAGCCAGC TCCTGCTGGC ATGGCCAGCA GCCAAGGGGG ACCAGCCCTC
  361 CTCCAGCCTG TTCCTGCTGA TGTGGTCAGC GCCCAGGGGG TACCATCCAT CCTCCAGCCA
  421 GCTCCTGCTG AGGTGATCAG CGCCCAAGCG ACACCACCCC TGCTCCAGCC TGCTCCGCAA
  481 CTGTCTGTTG ACCTGACAGA AGTGGAGGTC TTGGGAGAAG ACACTGTGGA GAACATCAAT
  541 CCAAGAACTT CAGAACAACA TAGGCAGGGA TCTGATGGTA ATCACACCAT CCCAGCATCT
  601 TCGTTGCATT CAATGACCAA CTTCATCAGC GGACTGCAGA GACTTCATGG CATGCTGGAA
  661 TTCCTGAGAC CTTCATCTTC AAACCACAGT GTAGGGCCAA TGAGAACAAG AAGGAGGGTA
  721 TCTGCTTCAC GGAGGGCAAG AGCCGGANGG TCTCAGAGGA NAGACAGTGC CAGGTTGAGA
  781 GCACCATTGG ATGCTTACTT TCAGGTGAGC AGGANCCAGC CTGACTTGCC NGCTACCACT
  841 TATGATTCAN AGACTANGAA TCCTGTATCT GAAGANTTGC ANGTGTCTAG TAGTTCTGAT
  901 TCTGANANTG NNAGCTCTGC ANANTATGGA NGGGTTGNTG NNCNNGCANA NGNNNCTGGA
961 NCTGNCATTT TANANNANCN NNNANCNNNN GNNNNANCAN ANCNNNNNNN NNN
//
Primer: BGH_rev2 (Pr0606)
Region: bGH p(A), attB2, insert 3'
Sequence file: RDB17493_A9Iob.seq check
>D06551A4_A9Io_1_BGH_rev2_C02_08_ABI24.ab1
    1 NNNNNNNNNN NNNNNNNNNG GNGGNNGGGC 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 NCAGTGGGCA TCTGGCTTTC TGAGCTACTA
  721 ACTCCTGCAC ATGACTGCTC GTGTTTCGCA CGTCATATAC CAGAATTGAA CCATTGGCCA
  781 GTCCAGCATA GATGTAGTTA GCCTCATCAA GACACCAGCA ACAGCTCCAG ACAGGANGTC
  841 CAGCATTATA AGTCTGGANC ACGGNATTTG TCTNCNGGCT GGTCAGTTTA ATAGTGNTGN
  901 CTANGGAAGC AGANAGTANC AAGCCTCTGA NNTNNCTGNT AAACGCCAGN NNNCGGATCN
961 GNTNGNNNTG CATCNGANNG TACTGACNGN TCNTCNTGNN NNNNNNNN
//
Primer: hRFWD3-N_F (Pr0753)
Region: insert mid
Sequence file: RDB17493_A9Ioc.seq check
>D06551A4_A9Io_1_hRFWD3-N_F_D04_10_ABI24.ab1
    1 NNNNNNNNNN NNNNNCNNNN ATCTGNNNNN TGCAGGTGTC TAGTAGTTCT GATTCTGACA
   61 GTGACAGCTC TGCAGAGTAT GGAGGGGTTG TTGACCAGGC AGAGGAATCT GGAGCTGTCA
  121 TTTTAGAAGA GCAACTAGCA GGTGTCTCAG CAGAGCAAGA AGTTACATGT ATCGATGGAG
  181 GCAAGACCCT CCCCAAACAG CCATCTCCCC AGAAGTCTGA GCCTCTGCTA CCTTCTGCTT
  241 CTATGGATGA GGAAGAAGGG GACACTTGTA CAATATGTCT GGAACAGTGG ACCAATGCTG
  301 GGGACCACCG GCTCTCAGCA TTACGCTGTG GGCATCTCTT TGGGTATAGG TGCATTTCCA
  361 CGTGGCTTAA AGGACAAGTA CGAAAATGTC CCCAGTGCAA CAAGAAAGCC AGGCACAGTG
  421 ACATTGTCGT CCTTTATGCC CGAACCCTGA GAGCTTTGGA CACTAGTGAA CAGGAGCGCA
  481 TGAAAAGTTC CCTACTGAAG GAACAGATGC TAAGGAAACA GGCCGAGTTA GAATCAGCAC
  541 AGTGCCGACT CCAACTGCAG GTCCTCACTG ATAAGTGCAC TAAGCTTCAA AGGCGTGTTC
  601 AGGACTTGCA AAAACTTACG TCACATCAAA GTCAGAATTT ACAGCAACCC AGGGGCTCCC
  661 AAGCATGGGT CCTGAGCTGC TCACCCTCCA GCCAGGGCCA GCACAAGCAC AAGTACCACT
  721 TCCAAAAGAC CTTCACAGTA TCTCAGGCAG GAAACTGCCG GATCATGGCA TACTGTGATG
  781 CTCTGAGCTG CCTGGTGATA TCACAGCCTT CTCCTCANGC CTCTTTNCTT CCAGGCTTTG
  841 GTGTTAAGAT GTTGAGTANT GCCAACATGA AGAGCAGTCN GTACATTCCG ATGCATGGCA
  901 AACAGATCCG NGNANTGNCG TTNANCAGTT ACCTCANANG CTTGCTACTC TNNGCTNNCN
  961 NANANNNCAC NATTAANNTG ANNAGCCNNN NNNNNAATAC CGNNGNNNNN ANNTATNNNG
1021 NNNNNNNNNN
//
Primer: SV40pro_ori_F (Pr0731)
Region: SV40 pro_ori, bleoR
Sequence file: RDB17493_A9Iod.seq check
>D06551A4_A9Io_1_SV40pro_ori_F_D02_11_ABI24.ab1
    1 NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN TGCNTCTGAG CTNTTCNNGA AGTAGTGAGG
   61 AGGCTTTTTT GGAGGCCTAG GCTTTTGCAA AAAGCTCCCG GGAGCTTGTA TATCCATTTT
  121 CGGATCTGAT CAGCACGTGT TGACAATTAA TCATCGGCAT AGTATATCGG CATAGTATAA
  181 TACGACAAGG TGAGGAACTA AACCATGGCC AAGTTGACCA GTGCCGTTCC GGTGCTCACC
  241 GCGCGCGACG TCGCCGGAGC GGTCGAGTTC TGGACCGACC GGCTCGGGTT CTCCCGGGAC
  301 TTCGTGGAGG ACGACTTCGC CGGTGTGGTC CGGGACGACG TGACCCTGTT CATCAGCGCG
  361 GTCCAGGACC AGGTGGTGCC GGACAACACC CTGGCCTGGG TGTGGGTGCG CGGCCTGGAC
  421 GAGCTGTACG CCGAGTGGTC GGAGGTCGTG TCCACGAACT TCCGGGACGC CTCCGGGCCG
  481 GCCATGACCG AGATCGGCGA GCAGCCGTGG GGGCGGGAGT TCGCCCTGCG CGACCCGGCC
  541 GGCAACTGCG TGCACTTCGT GGCCGAGGAG CANGACTGAC ACGTGCTACG AGATTTCGAT
  601 TCCACCGCCG CCTTCTATGA AAGGTTGGGC TTCGGAATCG TTTTCCGGGA CGCCGGCTGG
  661 ATGATCCTCC AGCGCGGGGA TCTCATGCTG GAGTTCTTCG CCCACCCCAA CTTGTTTATT
  721 GCAGCTTATA ATGGTTACAA ATAAAGCAAT AGCATCACAA ATTTCACAAA TAAAGCATTT
  781 TTTTCACTGC ATTCTAGTTG TGGTTTGTCC NAACTCATCN ATGTATCTTA TCATGTCTGT
  841 ATACCGTCGA CCTCTANCTA GAGCTTGGCG TANNCATGGT CATAGCTGTT TCCTGTGTGA
  901 AATTGTTANC CGCTCACANT TNCNCACAAC NTACGANCCN GNANNCNNNA NGTGNNNAAG
  961 CCNGGGGGTG NCNNNNNGAG TGAGCNNACT CACATTNANT TGCNNTTGCN CTNNNNNNCC
1021 NNNTTTNNNG NNNNGGGAAA NNNNNNNNNN
//

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]