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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).
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. |

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 literature designated by the DEPOSITOR is requested. (Morikawa, T.J. et al., Sci. Rep. 6: 22342, 2016)
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Schedule A (GFP-A) [Link]
Remarks
Please use Schedule A (GFP-A) additionally.
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Catalog # Resource name Shipping form Fee (non-profit org.)
RDB14204 pGimRET DNA solution

Ordering Information [in Japanese] [in English]
Please review the QC test results indicated by check icon below as well as clone information before placing your order.

References and tips

Electronic file

Original reference

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.
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.

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

Featured content

Featured content Dnaconda's recommendation EXR0059e (English text)
Featured content Dnaconda's recommendation EXR0059j (Japanese text)
Featured content Sensor and Visualization (English text)

Sequence information

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_A6Fop1-2.pdf check

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

Primer: mTurquoise2-M_F
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
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
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
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
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
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.
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.
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).

2021.08.05

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