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pGimRET

Expression vector of GimRET (Glycine inserted mutant FRET probe) for mammalian cells.

Catalog number RDB14204
Resource name pGimRET
Clone info. Expression vector of glycine-inserted mutant FRET (GimRET) fusion protein for Mammalian cells. GimRET works as a protein-crowding sensor that consists of CFP with YFP1G (one glycine insertion).
Comment Expression was confirmed by the depositor with microsopy. YFP1G (Inserted a Gly residue in the position of the 145th Tyr residue).
Vector backbone pECFP-N1 (Plasmid)
Size of vector backbone 5.9 kb
Selectable markers Kan^r (E. coli), Neo^r (mammaliancells)
Gene/insert name Aequorea victoria GFP cDNA
Depositor|Developer Watanabe, Tomonobu M. |
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External Database
Aequorea victoria GFP

            Reference sequence
              
             

            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 for distribution In publishing the research results obtained by use of the BIOLOGICAL RESOURCE, a citation of literature designated by the DEPOSITOR is requested. (Morikawa, T.J. et al., Sci. Rep. 6: 22342, 2016)
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            提供条件 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 (Morikawa, T.J. et al., Sci. Rep. 6: 22342, 2016)。
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            Catalog # Resource name Shipping form Fee (non-profit org.)
            RDB14204 pGimRET DNA solution

            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 pGimRET was provided by the RIKEN BRC through the National BioResource Project of the MEXT, Japan (cat. RDB14204).

            Reference section:

            Konishi, H.A., Asai, S., Watanabe, T.M., Yoshimura, S.H., In vivo analysis of protein crowding within the nuclear pore complex in interphase and mitosis. Sci. Rep. 7 (1): 5709 (2017). PMID 28720791. [link to RRC of NBRP]
            Morikawa, T.J., Fujita, H., Kitamura, A., Horio, T., Yamamoto, J., Kinjo, M., Sasaki, A., Machiyama, H., Yoshizawa, K., Ichimura, T., Imada, K., Nagai, T., Watanabe, T.M., Dependence of fluorescent protein brightness on protein concentration in solution and enhancement of it. Sci. Rep. 6: 22342 (2016). PMID 26956628. [link to RRC of NBRP]

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


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            Featured content

            Featured content GimRET for visualization of protein concentration in cells. (English text)
            Featured content GimRET for visualization of protein concentration in cells. (Japanese text)
            Featured content Sensor and Visualization (English text)

            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 RDB14204_A6Fop1-4.pdf check

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

            Primer: mTurquoise2-M_F (Pr0577)
            Region: ECFP 3',insert 5'(YFP1G 5')
            Sequence file: RDB14204_A6Fod.seq check
            >14204_14204_B1GgF_mTurquoise2-M_F_B01_02_ABI08.ab1
                1 NNNNNNNNNN NNCTCGCCGN NNNCTACCNG CAGAACACCC CCATCGGCGA CGGCCCCGTG
               61 CTGCTGCCCG ACAACCACTA CCTGAGCACC CAGTCCGCCC TGAGCAAAGA CCCCAACGAG
              121 AAGCGCGATC ACATGGTCCT GCTGGAGTTC GTGACCGCCG CCGGGATCAC TCTCGGCATG
              181 GACGAGCTGT ACAAGTCCGG AGGCAGCACC AGCGGTACTG GAGGTTCAGG CGGCTCAGGT
              241 GGAACTGGTG TGAGCAAGGG CGAGGAGCTG TTCACCGGGG TGGTGCCCAT CCTGGTCGAG
              301 CTGGACGGCG ACGTAAACGG CCACAAGTTC AGCGTGTCCG GCGAGGGCGA GGGCGATGCC
              361 ACCTACGGCA AGCTGACCCT GAAGTTCATC TGCACCACCG GCAAGCTGCC CGTGCCCTGG
              421 CCCACCCTCG TGACCACCTT CGGCTACGGC CTGCAGTGCT TCGCCCGCTA CCCCGACCAC
              481 ATGAAGCAGC ACGACTTCTT CAAGTCCGCC ATGCCCGAAG GCTACGTCCA GGAGCGCACC
              541 ATCTTCTTCA AGGACGACGG CAACTACNAG ACCCGCGCCG AGGTGAAGTT CGAGGGCGAC
              601 ACCCTGGTGN ACCGCATCGA GCTGNAGGGC ATCGANTTCA AGGAGGANGG CNACATNCNT
              661 GGGGCACAAG CTGGANTACN NCGGNNANAN AGCCNCNACG TCNATATCAN NNNNNANNNG
            721 CAGNNNNNNN GNNTCNNNNN NNANNNNNNN NNN
            //
            Primer: pAxEF R2 (Pr0059)
            Region: SV40 pA,insert 3'(YFP1G)
            Sequence file: RDB14204_A6Foe.seq check
            >14204_14204_B1GgF_pAxEF_R2_F03_18_ABI24.ab1
                1 NNNNNNNNNN NNTTNNNTTG TGNANNTTGT GANGCTATTG CTTTATTTGT AACCATTATA
               61 AGCTGCAATA AACAAGTTAA CAACAACAAT TGCATTCATT TTATGTTTCA GGTTCAGGGG
              121 GAGGTGTGGG AGGTTTTTTA AAGCAAGTAA AACCTCTACA AATGTGGTAT GGCTGATTAT
              181 GATCAGTTAT CTAGATCCGG TGGATCCCGG GCCCGCGGTA CCGTCGACTG CAGAATTCGA
              241 AGCTTGAGCT CGAGTTACTT GTACAGCTCG TCCATGCCGA GAGTGATCCC GGCGGCGGTC
              301 ACGAACTCCA GCAGGACCAT GTGATCGCGC TTCTCGTTGG GGTCTTTGCT CAGGGCGGAC
              361 TGGTAGCTCA GGTAGTGGTT GTCGGGCAGC AGCACGGGGC CGTCGCCGAT GGGGGTGTTC
              421 TGCTGGTAGT GGTCGGCGAG CTGCACGCTG CCGTCCTCGA TGTTGTGGCG GATCTTGAAG
              481 TTCACCTTGA TGCCGTTCTT CTGCTTGTCG GCCATGATAT ANACNNTGTG GCTGTTGTAT
              541 CCGTTGTACT CCNGCTTGNN CCCCANGATG TTGNNGTCCT CCTTGAANNN NNNNNNCCTN
              601 NNNCTCGNNG CGGNNNNNNN NNGNNGNCNC CNNNNAANNT NNNNNNCNNC GCGGGNNNTN
            661 GNANNTGCCG NCNNCNNTGN ANNANNANNN NGNNNNNNNN NN
            //
            Primer: CMV_Forward (Pr0016)
            Region: CMV pro,ECFP 5'
            Sequence file: RDB14204_A6Fof.seq check
            >14204_14204_B1GgF_CMV_Forward_A01_01_ABI08.ab1
                1 NNNNNNNNNN NNNNNNNGNN GCNNTGGGCG GTAGGCGTGT ACGGTGGGAG GTCTATATAA
               61 GCAGAGCTGG TTTAGTGAAC CGTCAGATCC GCTAGCGCTA CCGGTCGCCA CCATGGTGAG
              121 CAAGGGCGAG GAGCTGTTCA CCGGGGTGGT GCCCATCCTG GTCGAGCTGG ACGGCGACGT
              181 AAACGGCCAC AAGTTCAGCG TGTCCGGCGA GGGCGAGGGC GATGCCACCT ACGGCAAGCT
              241 GACCCTGAAG TTCATCTGCA CCACCGGCAA GCTGCCCGTG CCCTGGCCCA CCCTCGTGAC
              301 CACCCTGACC TGGGGCGTGC AGTGCTTCAG CCGCTACCCC GACCACATGA AGCAGCACGA
              361 CTTCTTCAAG TCCGCCATGC CCGAAGGCTA CGTCCAGGAG CGCACCATCT TCTTCAAGGA
              421 CGACGGCAAC TACAAGACCC GCGCCGAGGT GAAGTTCGAG GGCGACACCC TGGTGAACCG
              481 CATCGAGCTG AAGGGCATCG ACTTCAAGGA GGACGGCAAC ATCCTGGGGC ACAAGCTGGA
              541 GTACAACTAC ATCAGCCACA ACGTCTATAT CACCGCCGAC NAGCAGAAGA ACGGCATCAA
              601 GGCCAACTTC AAGATCCGCC ACAACATCGA GGANGGCAGC GTGCAGCTCG CCGANCACTA
            661 CCNGCANAAN ACCCCCNTCN GCGANNGNCC CNNNCTGCTG NCNNANNNNN NNNNN
            //
            Primer: FP-N_F1 (Pr0920)
            Region: ECFP 3',insert 5'(YFP1G 5')
            Sequence file: RDB14204_A6Fog.seq check
            >14204_14204_B1GgF_EP-N_F1_G01_07_ABI08.ab1
                1 NNNNNNNNNN NNNNGTCCGN CCATNNCCCG AAGGCTACGT CCNGGAGCGC ACCATCTTCT
               61 TCAAGGACGA CGGCAACTAC AAGACCCGCG CCGAGGTGAA GTTCGAGGGC GACACCCTGG
              121 TGAACCGCAT CGAGCTGAAG GGCATCGACT TCAAGGAGGA CGGCAACATC CTGGGGCACA
              181 AGCTGGAGTA CAACTACATC AGCCACAACG TCTATATCAC CGCCGACAAG CAGAAGAACG
              241 GCATCAAGGC CAACTTCAAG ATCCGCCACA ACATCGAGGA CGGCAGCGTG CAGCTCGCCG
              301 ACCACTACCA GCAGAACACC CCCATCGGCG ACGGCCCCGT GCTGCTGCCC GACAACCACT
              361 ACCTGAGCAC CCAGTCCGCC CTGAGCAAAG ACCCCAACGA GAAGCGCGAT CACATGGTCC
              421 TGCTGGAGTT CGTGACCGCC GCCGGGATCA CTCTCGGCAT GGACGAGCTG TACAAGTCCG
              481 GAGGCAGCAC CAGCGGTACT GGAGGTTCAG GCGGCTCAGG TGGAACTGGT GTGAGCAAGG
              541 GCGANGANCT GTTCACCGGG GTGGTGCCCN TCCTGNTCGA GCTGGACGGC GACGTAANCG
              601 GNCNCAAGTT CNGCGTGTCC GGCGANGGCN ANGNNGANGN CNNCNANNNN NNGCTGANCC
            661 TNNNGTTCNT CTGNNNNNNN GGNNNNCTGN CCNNNNCNNN NNCNNNCCNN NNNNNNNN
            //
            Primer: pGL3_RV5F (Pr0227)
            Region: insert 3'(YFP1G 3')
            Sequence file: RDB14204_A6Foh.seq check
            >D03467A2_A6Fo_1_pGL3_RV5F_A03_03.ab1
                1 NNNNNNNNNN NNNNNNNNGG CTGNNTANGN NNNGTTATCT AGATCCGGTG GNTCCCGGGC
               61 CCGCGGTACC GTCGACTGCA GAATTCGAAG CTTGAGCTCG AGTTACTTGT ACAGCTCGTC
              121 CATGCCGAGA GTGATCCCGG CGGCGGTCAC GAACTCCAGC AGGACCATGT GATCGCGCTT
              181 CTCGTTGGGG TCTTTGCTCA GGGCGGACTG GTAGCTCAGG TAGTGGTTGT CGGGCAGCAG
              241 CACGGGGCCG TCGCCGATGG GGGTGTTCTG CTGGTAGTGG TCGGCGAGCT GCACGCTGCC
              301 GTCCTCGATG TTGTGGCGGA TCTTGAAGTT CACCTTGATG CCGTTCTTCT GCTTGTCGGC
              361 CATGATATAG ACGTTGTGGC TGTTGTATCC GTTGTACTCC AGCTTGTGCC CCAGGATGTT
              421 GCCGTCCTCC TTGAAGTCGA TGCCCTTCAG CTCGATGCGG TTCACCAGGG TGTCGCCCTC
              481 GAACTTCACC TCGGCGCGGG TCTTGTAGTT GCCGTCGTCC TTGAAGAAGA TGGTGCGCTC
              541 CTGGACGTAG CCTTCGGGCA TGGCGGACTT GAAGAAGTCG TGCTGCTTCA TGTGGTCGGG
              601 GTAGCGGGCG AAGCACTGCA GGCCGTAGCC GAAGGTGGTC ACGAGGGTGG GCCAGGGCAC
              661 GGGCAGCTTG CCGGTGGTGC AGATGAACTT CAGGGTCAGC TTGCCGTANG TGGCATCGCC
              721 CTCGCCCTCG CCGGACACGC TGAACTTGTG GCCGTTTACG TCGCCGTCCA GCTCGACCAG
              781 GATGGGCACC ACCCCGGTGA ACAGCTCCTC GCCCTTNCTC ACACCAGTTC CACCTGANCC
              841 GCCTGAACCT CCAGTACCGC TGGTGCTGCC NNCGGANTTG TACAGCTCGN NCNNGCCNAN
              901 ANTGNTNCNN NCGGNNGNCA CGAACTCCNG CANGANCATG NNATCGCGCT TNNCNTTNGG
              961 GGNCTTTGNN NNNNNCNNNN TNGGGNGCTN NNNNTAGTNG NTNGNNNGGN NNNNNCNNNG
            1021 GGNNNNNNNN NN
            //
            Primer: SV40pro_F_V2 (Pr0154)
            Region: KanR_NeoR
            Sequence file: RDB14204_A6Foi.seq check
            >D03467A2_A6Fo_1_SV40pro_F_V2_B03_06.ab1
                1 NNNNNNNNNN NNGNNCNNNG CTTTTGCAAG NNCGNTCNAG AGACAGGNNG AGGNTCGTTT
               61 CGCATGATTG AACAAGATGG ATTGCACGCA GGTTCTCCGG CCGCTTGGGT GGAGAGGCTA
              121 TTCGGCTATG ACTGGGCACA ACAGACAATC GGCTGCTCTG ATGCCGCCGT GTTCCGGCTG
              181 TCAGCGCAGG GGCGCCCGGT TCTTTTTGTC AAGACCGACC TGTCCGGTGC CCTGAATGAA
              241 CTGCAAGACG AGGCAGCGCG GCTATCGTGG CTGGCCACGA CGGGCGTTCC TTGCGCAGCT
              301 GTGCTCGACG TTGTCACTGA AGCGGGAAGG GACTGGCTGC TATTGGGCGA AGTGCCGGGG
              361 CAGGATCTCC TGTCATCTCA CCTTGCTCCT GCCGAGAAAG TATCCATCAT GGCTGATGCA
              421 ATGCGGCGGC TGCATACGCT TGATCCGGCT ACCTGCCCAT TCGACCACCA AGCGAAACAT
              481 CGCATCGAGC GAGCACGTAC TCGGATGGAA GCCGGTCTTG TCGATCAGGA TGATCTGGAC
              541 GAAGAGCATC AGGGGCTCGC GCCAGCCGAA CTGTTCGCCA GGCTCAAGGC GAGCATGCCC
              601 GACGGCGAGG ATCTCGTCGT GACCCATGGC GATGCCTGCT TGCCGAATAT CATGGTGGAA
              661 AATGGCCGCT TTTCTGGATT CATCGACTGT GGCCGGCTGG GTGTGGCGGA NCGCTATCAG
              721 GACATAGCGT TGGCTACCCG TGATATTGCT GAAGAGCTTG GCGGCGAATG GGCTGACCGC
              781 TTCCTCGTGC TTTACGGTAT CGCCGCTCCC GATTCGCAGC GCATCGCCTT CTATCGCCTT
              841 CTTGACGANT TCTTCTGANC GGGNNTCNNG GGTTCGAAAT GNNCGANNNA NCGACGCCCN
              901 ACCTGCCATC ACGANATTTC NANTCCNCCG CCGCCNTCTA NNAANNNNTG GGCTTNNNNN
            961 NNNNTTTNCN NGNNNNNCNN NNNNNNNN
            //

            Please visit Sequencing and PCR primers for primer information.


            References

            Original, user report and related articles

            original Konishi, H.A., In vivo analysis of protein crowding within the nuclear pore complex in interphase and mitosis. Sci. Rep. 7 (1): 5709 (2017). PMID 28720791. [link to RRC of NBRP]
            original Morikawa, T.J., Dependence of fluorescent protein brightness on protein concentration in solution and enhancement of it. Sci. Rep. 6: 22342 (2016). PMID 26956628. [link to RRC of NBRP]
            reference Machiyama, H., The use of a genetically encoded molecular crowding sensor in various biological phenomena. Biophys. Physicobiol. 14: 119-125 (2017). PMID 28900589.
            review Watanabe, T.M., An intracellular crowding sensor based on fluorescent protein. (Japanese text) Journal of Japanese Biochemical Society 89 (4): 577-586 (2017).

            2024.03.26

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