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pPAL7_GimRET (#RDB14203)

Expression vector of GimRET (Glycine inserted mutant FRET probe) for E. coli.

Alternative name CFP-YFP1G GimRET
Clone info. Expression vector of glycine-inserted mutant FRET (GimRET) fusion protein for E.coli. GimRET works as a protein-crowding sensor that consists of CFP with YFP1G (one glycine insertion).
Comment YFP1G (Inserted a Gly residue in the position of the 145th Tyr residue).
Vector backbone pAL7 (Plasmid)
Size of vector backbone 5.9 kb
Selectable markers Amp^r
Gene/insert name Aequorea victoria GFP cDNA
Depositor|Developer Watanabe, Tomonobu |
 

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. (Morikawa, T.J. et al., Sci. Rep. 6: 22342, 2016)
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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 (Morikawa, T.J. et al., Sci. Rep. 6: 22342, 2016)。

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RDB14203 pPAL7_GimRET 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 pPAL7_GimRET was provided by the RIKEN BRC through the National BioResource Project of the MEXT, Japan (cat. RDB14203).

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)

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 RDB14203_A6Fop1-2.pdf check

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

Primer: pBR322_BamHI_R (Pr0065)
Region: lac O,RBS,Profinity eXact tag,ECFP 5'
Sequence file: RDB14203_A6Fod.seq check
>D03467A1_A6Fo_1_pBR322BamHI_R_G02_20.ab1
    1 NNNNNNNNNN NNNNNNNNNN NNNNNNNTNG GGNNNNNNTG AGCGGNNAAC AATTCCCCTC
   61 TAGAAATAAT TTTGTTTAAC TTTAAGAAGG AGATATACAT ATGGGAGGGA AATCAAACGG
  121 GGAAAAGAAA TATATTGTCG GGTTCAAACA GGGCTTTAAG AGCTGCGCTA AGAAGGAGGA
  181 TGTCATTTCT GAAAAAGGCG GGAAACTCCA AAAGTGCTTC AAATATGTAG ACGCAGCTAG
  241 CGCTACATTA AACGAAAAAG CTGTAGAAGA ATTGAAAAAA GATCCGAGCG TCGCGTACGT
  301 AGAAGAAGAC AAGCTCTTCA AAGCTTTGGT GAGCAAGGGC GAGGAGCTGT TCACCGGGGT
  361 GGTGCCCATC CTGGTCGAGC TGGACGGCGA CGTAAACGGC CACAAGTTCA GCGTGTCCGG
  421 CGAGGGCGAG GGCGATGCCA CCTACGGCAA GCTGACCCTG AAGTTCATCT GCACCACCGG
  481 CAAGCTGCCC GTGCCCTGGC CCACCCTCGT GACCACCCTG ACCTGGGGCG TGCAGTGCTT
  541 CAGCCGCTAC CCCGACCACA TGAAGCAGCA CGACTTCTTC AAGTCCGCCA TGCCCGAAGG
  601 CTACGTCCAG GANCGCACCA TCTTCTTCAA GGACGACNGC AACTACNAGA NCCGCGCCGA
  661 NGTGAAGTTC GAGGGCGACA CCCTGGTGAA CCGCATCGAG CTGAANGGCA TCGACTTCAN
  721 NGANGANGGC AACATCCTGG GGCACNANCT GNANTANANC TACNTCAGCN NNNACGTCTA
  781 TATCNCCGCC GANNAGCNNA ANANNGGCAT NNAGGCNNNN TTCNNNATNC NNCCACNNNN
  841 TCNNNGANNG NANNNNNGCA NNTNNNNNAN NNNTNCNNNN NNANNNNCCC CNNTCGGCGN
901 NGNNCCNNNN NNNNNN
//
Primer: T7ter_R1 (Pr0798)
Region: EYFP 3'(YFP1G)
Sequence file: RDB14203_A6Foe.seq check
>14203_14203_B1H2F_T7ter_R1_A03_03_ABI24.ab1
    1 NNNNNNNNNN NNNNNNNNNN NNNNTTGCTN NGCGGNGGCA GCNNGCCAAC TCAGCTTCCT
   61 TTCGGGCTTA TGCGGCCGCC TCGAGTTACT TGTACAGCTC GTCCATGCCG AGAGTGATCC
  121 CGGCGGCGGT CACGAACTCC AGCAGGACCA TGTGATCGCG CTTCTCGTTG GGGTCTTTGC
  181 TCAGGGCGGA CTGGTAGCTC AGGTAGTGGT TGTCGGGCAG CAGCACGGGG CCGTCGCCGA
  241 TGGGGGTGTT CTGCTGGTAG TGGTCGGCGA GCTGCACGCT GCCGTCCTCG ATGTTGTGGC
  301 GGATCTTGAA GTTCACCTTG ATGCCGTTCT TCTGCTTGTC GGCCATGATA TAGACGTTGT
  361 GGCTGTTGTA TCCGTTGTAC TCCAGCTTGT GCCCCAGGAT GTTGCCGTCC TCCTTGAAGT
  421 CGATGCCCTT CAGCTCGATG CGGTTCACCA GGGTGTCGCC CTCGAACTTC ACCTCGGCGC
  481 GGGTCTTGTA GTTGCCGTCG TCCTTGAAGA AGATGGTGCG CTCCTGGACG TAGCCTTCGG
  541 GCATGGCGGA CTTGAAGAAG TCGTGCTGCT TCATGTGGTC GGGGTAGCGG GCGAAGCACT
  601 GCANGCCGTA NCCGAANGTG GTCACGAGGG TGGGCCAGGG NACGGGCAGC TTGNCGGTGG
  661 TGCANANGNA NNTCNNNNNC AGCTNGNCGT NNNGGNATCN NCCNCNNCNN CNNNNGNANA
721 CNCNNAANTT NNGGNNNNTN NNNNNNNNCN NNNNN
//
Primer: EYFP-H_R (Pr1036)
Region: ECFP mid
Sequence file: RDB14203_A6Fof.seq check
>14203_14203_B1H2F_EYFP-H_R_B03_06_ABI24.ab1
    1 NNNNNNNNNN NNNNNNNNNN NNNNNGCTGG NGCTGCCTCC GGNCTTGTAC NGCTCGTCCA
   61 TGCCGAGAGT GATCCCGGCG GCGGTCACGA ACTCCAGCAG GACCATGTGA TCGCGCTTCT
  121 CGTTGGGGTC TTTGCTCAGG GCGGACTGGG TGCTCAGGTA GTGGTTGTCG GGCAGCAGCA
  181 CGGGGCCGTC GCCGATGGGG GTGTTCTGCT GGTAGTGGTC GGCGAGCTGC ACGCTGCCGT
  241 CCTCGATGTT GTGGCGGATC TTGAAGTTGG CCTTGATGCC GTTCTTCTGC TTGTCGGCGG
  301 TGATATAGAC GTTGTGGCTG ATGTAGTTGT ACTCCAGCTT GTGCCCCAGG ATGTTGCCGT
  361 CCTCCTTGAA GTCGATGCCC TTCAGCTCGA TGCGGTTCAC CAGGGTGTCG CCCTCGAACT
  421 TCACCTCGGC GCGGGTCTTG TAGTTGCCGT CGTCCTTGAA GAAGATGGTG CGCTCCTGGA
  481 CGTAGCCTTC GGGCATGGCG GACTTGAAGA AGTCGTGCTG CTTCATGTGG TCGGGGTAGC
  541 GGCTGAAGCA CTGCACGCCC CAGGTCANGG TGGTCACGAN GGTGGGCCAG GGCACGGGCA
  601 GCTTGCCGGT GGTGCAGATG AACTTCAGGG TCNNCTTGCC GNNNNGGCAT CGCCCTCNCC
661 CTCNNCNGAC NNNCTNNNNN NNNNN
//
Primer: ECFP-C_F1 (Pr0925)
Region: ECFP 3',EYFP(YFP1G) 5'
Sequence file: RDB14203_A6Fog.seq check
>14203_14203_B1H2F_ECFP-C_F1_C03_09_ABI24.ab1
    1 NNNNNNNNNN NNNNNNNNNN NNNCNNNNNN CNTGGTCCTG CTGGAGTTCG TGACCGCCGC
   61 CGGGATCACT CTCGGCATGG ACGAGCTGTA CAAGTCCGGA GGCAGCACCA GCGGTACTGG
  121 AGGTTCAGGC GGCTCAGGTG GAACTGGTGT GAGCAAGGGC GAGGAGCTGT TCACCGGGGT
  181 GGTGCCCATC CTGGTCGAGC TGGACGGCGA CGTAAACGGC CACAAGTTCA GCGTGTCCGG
  241 CGAGGGCGAG GGCGATGCCA CCTACGGCAA GCTGACCCTG AAGTTCATCT GCACCACCGG
  301 CAAGCTGCCC GTGCCCTGGC CCACCCTCGT GACCACCTTC GGCTACGGCC TGCAGTGCTT
  361 CGCCCGCTAC CCCGACCACA TGAAGCAGCA CGACTTCTTC AAGTCCGCCA TGCCCGAAGG
  421 CTACGTCCAG GAGCGCACCA TCTTCTTCAA GGACGACGGC AACTACAAGA CCCGCGCCGA
  481 GGTGAAGTTC GAGGGCGACA CCCTGGTGAA CCGCATCGAG CTGAAGGGCA TCGACTTCAA
  541 GGAGGACGGC AACATCCTGG GGCACAAGCT GGAGTACAAC GGATACAACA GCCACNACGT
  601 CTATATCATG GCCGACAAGC AGAANAACNG CATCNAGGTG AACTTCNAGA NNCGCCNCNA
  661 CATCGAGGAN NGCAGNNTGC ANCTCNNNGA NCNCTACCAG CAGANNACCC CNTCNGNCGA
721 NNNNCNNNNN NNNNN
//
Primer: lacIpro_F3 (Pr0722)
Region: lacI pro,lacI
Sequence file: RDB14203_A6Foh.seq check
>14203_14203_B1H2F_laclpro_F3_D03_12_ABI24.ab1
    1 NNNNNNNNNN NNNNNNNNNN NNCTGGGCTG CTTCCTAATG CAGGAGTCGC ATAAGGGAGA
   61 GCGTCGAGAT CCCGGACACC ATCGAATGGC GCAAAACCTT TCGCGGTATG GCATGATAGC
  121 GCCCGGAAGA GAGTCAATTC AGGGTGGTGA ATGTGAAACC AGTAACGTTA TACGATGTCG
  181 CAGAGTATGC CGGTGTCTCT TATCAGACCG TTTCCCGCGT GGTGAACCAG GCCAGCCACG
  241 TTTCTGCGAA AACGCGGGAA AAAGTGGAAG CGGCGATGGC GGAGCTGAAT TACATTCCCA
  301 ACCGCGTGGC ACAACAACTG GCGGGCAAAC AGTCGTTGCT GATTGGCGTT GCCACCTCCA
  361 GTCTGGCCCT GCACGCGCCG TCGCAAATTG TCGCGGCGAT TAAATCTCGC GCCGATCAAC
  421 TGGGTGCCAG CGTGGTGGTG TCGATGGTAG AACGAAGCGG CGTCGAAGCC TGTAAAGCGG
  481 CGGTGCACAA TCTTCTCGCG CAACGCGTCA GTGGGCTGAT CATTAACTAT CCGCTGGATG
  541 ACCAGGATGC CATTGCTGTG GAAGCTGCCT GCACTAATGT TCCGGCGTTA TTTCTTGATG
  601 TCTCTGACCA GACACCCATC AACAGTATTA TTTTCTCCCN TGAANANGGN ACGCGACTGG
  661 GCGNNNNNCA TCTGNTCGCA TTGGGTCANC AGCAANTCGN GCTNNNNNCG GGNCNNNNNN
721 NNNN
//

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).