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pSFL61 pRS316-GPDp-Ifa38-mCherry-GFP11

Expression vector of S. cerevisiae ketoreductase (Ifa38) fused with mCherry-GFP11 at C-terminus.

Catalog number RDB18572
Resource name pSFL61 pRS316-GPDp-Ifa38-mCherry-GFP11
Clone info. The split-GFP system clone for visualizing organelle contact sites in vivo. Expression vector of S. cerevisiae ketoreductase (Ifa38) fused with mCherry-GFP11 at C-terminus, GPD promoter, CYC1 terminator, please refer to Fig.1 and Table S1 of Kakimoto, Y. et al. Sci. Rep., 8 (1): 6175, 2018.
Expression was confirmed by the depositor with fluorescence microscopy.
Vector backbone pRS316 (plasmid)
Selectable markers Ampicillin (E. coli), URA3 (S. cerevisiae).
Gene/insert name S. cerevisiae IFA38 cDNA Aequorea victoria GFP cDNA Discosoma sp RFP cDNA
Depositor|Developer Tamura, Yasushi |

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. (Kakimoto, Y. et al. Sci. Rep. 8:6175, 2018)
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Material Transfer Agreement (MTA for use for not-for-profit academic purpose) [Word]
Schedule A (GFP-A) [Link]
Remarks
Please use Schedule A (GFP-A) additionally.
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RDB18572 pSFL61 pRS316-GPDp-Ifa38-mCherry-GFP11 DNA solution Please contact us.

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 Kakimoto, Y., Visualizing multiple inter-organelle contact sites using the organelle-targeted split-GFP system. Sci. Rep. 8 (1): 6175 (2018). PMID 29670150.

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

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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 RDB18572_B0J5p1-1.pdf check

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

Primer: HB-NOS_R
Region: T7 pro, insert 5' (GPD pro, Ifa38 5')
Sequence file: RDB18572_B0J5a.seq check
>D07393A6_B0J5_2-HB-NOS_R_A04_01_ABI24.ab1
    1 NNNNNNNNNN NNNNNNNNNN NNNGGGTNNN GCCNGGGTTT TCCCNGTCNC GACGTTGTAA
   61 AACGACGGCC AGTGAATTGT AATACGACTC ACTATAGGGC GAATTGGAGC TCAGTTTATC
  121 ATTATCAATA CTCGCCATTT CAAAGAATAC GTAAATAATT AATAGTAGTG ATTTTCCTAA
  181 CTTTATTTAG TCAAAAAATT AGCCTTTTAA TTCTGCTGTA ACCCGTACAT GCCCAAAATA
  241 GGGGGCGGGT TACACAGAAT ATATAACATC GTAGGTGTCT GGGTGAACAG TTTATTCCTG
  301 GCATCCACTA AATATAATGG AGCCCGCTTT TTAAGCTGGC ATCCAGAAAA AAAAAGAATC
  361 CCAGCACCAA AATATTGTTT TCTTCACCAA CCATCAGTTC ATAGGTCCAT TCTCTTAGCG
  421 CAACTACAGA GAACAGGGGC ACAAACAGGC AAAAAACGGG CACAACCTCA ATGGAGTGAT
  481 GCAACCTGCC TGGAGTAAAT GATGACACAA GGCAATTGAC CCACGCATGT ATCTATCTCA
  541 TTTTCTTACA CCTTCTATTA CCTTCTGCTC TCTCTGATTT GGAAAAAGCT GAAAAAAAAG
  601 GTTGAAACCA GTTCCCTGAA ATTATTCCCC TACTTGACTA ATAAGTATAT AAAGACGGTA
  661 GGTATTGATT GTAATTCTGT AAATCTATTT CTTAAACTTC TTAAATTCTA CTTTTATAGT
  721 TAGTCTTTTT TTTAGTTTTA AAACACCAGA ACTTAGTTTC GACGGATGCG GCCGCTCTAG
  781 AACTAGTGGA TCCATGACTT TTATGCAACA GCTTCNAGAN GCTGGGGNNA GANTTAGGTG
  841 TATCAATGGT CTTTTATGGG TTGTTTTTGG GCTTGGTGTC TTAAAATGTA CAACGNTATC
  901 ACTTANATTT TTANCTCTTA TTTTTTGATC TTTTTTTTAC TANNANCANN CNANTTNCNA
961 NNNNNATNGN NGNNNAAACN NGNNAAATAC NGNGCNATCN NNNGNNGNNN NNNNNNN
//
Primer: SV40pA3'Element Reverse
Region: T3 pro, insert 3' (CYC1 ter)
Sequence file: RDB18572_B0J5b.seq check
>D07393A6_B0J5_2_SV40pA3'ElementReverse_B01_02_ABI08.ab1
    1 NNNNNNNNNN NNNNNNNNNN NNNNGNAANN GCNNGCAACG CGGCCTTTTT ACGGTTCCTG
   61 GCCTTTTGCT GGCCTTTTGC TCACATGTTC TTTCCTGCGT TATCCCCTGA TTCTGTGGAT
  121 AACCGTATTA CCGCCTTTGA GTGAGCTGAT ACCGCTCGCC GCAGCCGAAC GACCGAGCGC
  181 AGCGAGTCAG TGAGCGAGGA AGCGGAAGAG CGCCCAATAC GCAAACCGCC TCTCCCCGCG
  241 CGTTGGCCGA TTCATTAATG CAGCTGGCAC GACAGGTTTC CCGACTGGAA AGCGGGCAGT
  301 GAGCGCAACG CAATTAATGT GAGTTAGCTC ACTCATTAGG CACCCCAGGC TTTACACTTT
  361 ATGCTTCCGG CTCGTATGTT GTGTGGAATT GTGAGCGGAT AACAATTTCA CACAGGAAAC
  421 AGCTATGACC ATGATTACGC CAAGCTCGAA ATTAACCCTC ACTAAAGGGA ACAAAAGCTG
  481 GTACCGGCCG CAAATTAAAG CCTTCGAGCG TCCCAAAACC TTCTCAAGCA AGGTTTTCAG
  541 TATAATGTTA CATGCGTACA CGCGTCTGTA CAGAAAAAAA AGAAAAATTT GAAATATAAA
  601 TAACGTTCTT AATACTAACA TAACTATAAA AAAATAAATA GGGACCTAGA CTTCAGGTTG
  661 TCTAACTCCT TCCTTTTCGG TTAGAGCGGA TGTGGGGGGA GGGCGTGAAT GTAAGCGTGA
  721 CATAACTAAT TACATGACTC GAGGTCGANT CAAGTTATGC CGGCAGCATT GACGTACTCG
  781 TGCAACACCA TGTGATCCCT GCTTGTACTC CCTCCNCCTG ANCCACCAGA TCTCTNGTAC
  841 AGTTCGTCCA TTNCACCCNN ANANNGTCTG NNNNCNGCTC NTTCATACNG NNNCNNNANN
  901 GNGNAANCTN CNNNGNGGNN ANNNANNTNN NGCTNGATGT NCACATTATA CNNNNCNNNN
961 NNNNNN
//
Primer: CYC1ter_R
Region: insert mid (CYC1 ter, GFP11, mCherry)
Sequence file: RDB18572_B0J5c.seq check
>D07393A6_B0J5_2-CYC1Ter_R_C04_07_ABI24.ab1
    1 NNNNNNNNNN NNNNNNNNNN GGNNGNGGGG GGNNGGCGTG NATGTAAGCG TGACATAACT
   61 AATTACATGA CTCGAGGTCG ACTCAAGTTA TGCCGGCAGC ATTGACGTAC TCGTGCAACA
  121 CCATGTGATC CCTGCTTGTA CTCCCTCCGC CTGACCCACC AGATCTCTTG TACAGTTCGT
  181 CCATTCCACC CGTAGAATGT CTGCCTTCGG CTCGTTCATA CTGTTCCACA ATGGTGTAAT
  241 CTTCATTGTG GCTAGTGATA TCCAGCTTGA TGTTCACATT ATACGCGCCT GGAAGCTGAA
  301 CGGGTTTCTT TGCCTTGTAG GTGGTCTTGA CTTCGGCATC GTAATGGCCT CCATCCTTCA
  361 GTTTCAGGCG CTGTTTGATT TCGCCCTTGA GTGCACCATC TTCGGGATAC ATACGCTCTG
  421 AGCTGGCTTC CCATCCCATC GTTTTCTTCT GCATCACTGG CCCGTCGGAA GGAAAGTTGG
  481 TTCCTCTCAG CTTCACCTTG TAGATGAACT CACCGTCTTG GAGGGAACTG TCCTGAGTCA
  541 CTGTGACAAC TCCTCCGTCC TCGAAGTTCA TGACCCTCTC CCATTTGAAC CCCTCGGGAA
  601 AGGAGAGTTT CAGGTAGTCT GGAATGTCCG CAGGATGTTT CACGTATGCC TTGCTCCCAT
  661 ACATGAACTG GGGAGACAGA ATGTCCCAGG CAAACGGCAG AGGTCCTCCT TTTGTCACCT
  721 TGAGTTTAGC GGTTTGTGTC CCCTCATATG GNCGGCCTTC CCCTTCACCC TCAATCTCNA
  781 ACTCGTGTCC ATTGACACTG CCCTCCATGT GTACCTTGAA CCGCATAAAC TCCTTTATGA
  841 NGGCCATGTT ATCCTCCTCA CCCNTGCNAA CGCCGCCNGC GGAAAGCTTT TCCTTTTTNA
  901 NCTGTCNNGC GGCTTTTTTT TANNGNNNTA ANTCTGATAN ANTTATGGAA NGAANANTTN
961 NNGGNANTTA ANNNTCNNNN ANTAAANNNC NNAANNNNNN NNNN
//
Primer: URA3pro_F
Region: URA3 pro, URA3
Sequence file: RDB18572_B0J5d.seq check
>D07393A6_B0J5_2_URA3pro_F_C01_03_ABI08.ab1
    1 NNNNNNNNNN NNNNNNANNN NNNANNTGCC NNGTATTCTT AACCCAACTG CACAGAACAA
   61 AAACCTGCAG GAAACGAAGA TAAATCATGT CGAAAGCTAC ATATAAGGAA CGTGCTGCTA
  121 CTCATCCTAG TCCTGTTGCT GCCAAGCTAT TTAATATCAT GCACGAAAAG CAAACAAACT
  181 TGTGTGCTTC ATTGGATGTT CGTACCACCA AGGAATTACT GGAGTTAGTT GAAGCATTAG
  241 GTCCCAAAAT TTGTTTACTA AAAACACATG TGGATATCTT GACTGATTTT TCCATGGAGG
  301 GCACAGTTAA GCCGCTAAAG GCATTATCCG CCAAGTACAA TTTTTTACTC TTCGAAGACA
  361 GAAAATTTGC TGACATTGGT AATACAGTCA AATTGCAGTA CTCTGCGGGT GTATACAGAA
  421 TAGCAGAATG GGCAGACATT ACGAATGCAC ACGGTGTGGT GGGCCCAGGT ATTGTTAGCG
  481 GTTTGAAGCA GGCGGCAGAA GAAGTAACAA AGGAACCTAG AGGCCTTTTG ATGTTAGCAG
  541 AATTGTCATG CAAGGGCTCC CTATCTACTG GAGAATATAC TAAGGGTACT GTTGACATTG
  601 CGAAGAGCGA CAAAGATTTT GTTATCGGCT TTATTGCTCA AAGAGACATG GGTGGAAGAG
  661 ATGAAGGTTA CGATTGGTTG ATTATGACAC CCGGTGTGGG TTTAGATGAC AAGGGAGACG
  721 CATTGGGTCA ACAGTATAGA ACCGTGGATG ANGTGGTCTC TACAGGNTCT GACATTATTA
  781 TTGTTGGAAG AGGANTATTT GCAAAGGGAA NGGANGCTAN NGTAGAGGGT GAACGTTACA
  841 GAAAAGCANG CTGGGAANCA TATTNGANAA NANGCGGCCN GCAAAACTNA AAAAACTGNA
  901 TTATNAGTAA ATGCNNGTAN ACTAAACTCN CAANNTANAG CTTNNNTTTN ANTATATCAG
961 TTANNACCCT NNGCNGNNGN GAANNNCNNN NNNNNNN
//
Primer: TDH3pro_F
Region: insert mid (GPD pro, Ifa38 5')
Sequence file: RDB18572_B0J5e.seq check
>D07393A6_B0J5_2_TDH3pro_F_E01_05_ABI08.ab1
    1 NNNNNNNNNN NNNNNGNNTG NNNTCTGTNA NTCTATTTCT TAAACTTCTT AAATTCTACT
   61 TTTATAGTTA GTCTTTTTTT TAGTTTTAAA ACACCAGAAC TTAGTTTCGA CGGATGCGGC
  121 CGCTCTAGAA CTAGTGGATC CATGACTTTT ATGCAACAGC TTCAAGAGGC TGGGGAAAGA
  181 TTTAGGTGTA TCAATGGTCT TTTATGGGTT GTTTTTGGGC TTGGTGTCTT AAAATGTACA
  241 ACGTTATCAC TTAGATTTTT AGCTCTTATT TTTGATCTTT TTTTACTACC AGCAGTCAAT
  301 TTCGACAAGT ATGGTGCTAA AACTGGTAAA TACTGTGCTA TCACTGGTGC AAGTGACGGT
  361 ATTGGTAAAG AATTTGCTAG ACAAATGGCG AAACGTGGCT TCAACTTGGT ATTGATCTCG
  421 AGAACGCAAT CTAAATTGGA GGCTTTACAA AAAGAACTAG AAGATCAACA TCATGTCGTT
  481 GTAAAGATTC TAGCCATTGA CATTGCGGAG GATAAAGAAT CCAACTATGA ATCCATTAAG
  541 GAATTGTGTG CGCAGTTACC AATCACCGTT TTGGTCAATA ATGTTGGTCA ATCACACTCC
  601 ATTCCTGTTC CATTTTTGGA AACAGAAGAG AAGGAGCTTA GAAATATTAT CACTATCAAT
  661 AACACTGCTA CATTATTAAT TACACAAATC ATTGCACCAA AGATTGTGGA AACTGTGAAA
  721 GCTGANAACA AGAAGTCGGG TACTCGTGGT TTGATCTTAA CCATGGGCTC ATTTGGTGGT
  781 CTGATTCCCA CCCCACTTTT GGCTACATAC AGNGGTTCGA AATCATTCTT ACAAGGTTGG
  841 TCTAACTCTN NGGCTGGGGA ANTATCTAAA GATGCTATCN ATGTNGAATT ANTCATTTCA
  901 TATTTGGNCA CTANCTCNAN GTCTAAAANC ANAANNNCAT CNTNNANGAT CCNNAANNNN
961 NNNNNN
//
Primer: Ifa38-C_F
Region: insert mid (Ifa38 3', mCherry 5')
Sequence file: RDB18572_B0J5f.seq check
>D07393A6_B0J5_2_Ifa38_C-F_B10_02_ABI08.ab1
    1 NNNNNNNNNN NNNNNNNNNN NTNNNGNNNN CTTTGGCGTT NANTCTAAGA NNTGTNAACT
   61 CCNTTAATTA TTCCTTCCNT AAATCTATCA GAATTAGAGC CTTAAAAAAA GCCGCAAGAC
  121 AGGTTAAAAA GGAAAAGCTT TCCGCTGGCG GCGTTAGCAA GGGTGAGGAG GATAACATGG
  181 CCATCATAAA GGAGTTTATG CGGTTCAAGG TACACATGGA GGGCAGTGTC AATGGACACG
  241 AGTTTGAGAT TGAGGGTGAA GGGGAAGGCC GGCCATATGA GGGGACACAA ACCGCTAAAC
  301 TCAAGGTGAC AAAAGGAGGA CCTCTGCCGT TTGCCTGGGA CATTCTGTCT CCCCAGTTCA
  361 TGTATGGGAG CAAGGCATAC GTGAAACATC CTGCGGACAT TCCAGACTAC CTGAAACTCT
  421 CCTTTCCCGA GGGGTTCAAA TGGGAGAGGG TCATGAACTT CGAGGACGGA GGAGTTGTCA
  481 CAGTGACTCA GGACAGTTCC CTCCAAGACG GTGAGTTCAT CTACAAGGTG AAGCTGAGAG
  541 GAACCAACTT TCCTTCCGAC GGGCCAGTGA TGCAGAAGAA AACGATGGGA TGGGAAGCCA
  601 GCTCAGAGCG TATGTATCCC GAAGATGGTG CACTCAAGGG CGAAATCAAA CAGCGCCTGA
  661 AACTGAAGGA TGGAGGCCAT TACGATGCCG AAGTCAAGAC CACCTACAAG GCAAAGAAAC
  721 CCGTTCAGCT TCCAGGCGCG TATAATGTGA ACATCAAGCT GGATATCACT AGCCACAATG
  781 AAGATTACAC CATTGTGGAN CAGTATGAAC GANCCGAANG CAGACATTCT ACGGGTGGAA
  841 TGGACGAACT GTACAAGAGA TCTGGTGGGT CANGCGGAGG GAGTACAAGC AGGNNTCACA
  901 TGGNGTTGCA CGANTACGTC NNNGCTGCCN NCATAACNNN ANTCGANCNN NNNNCATGTA
961 ATNANNTANG NNNCNNNNAC NTTNNNNNCN NNCCCCNNNN NNNNNN
//
Primer: Amp_R
Region: AmpR pro, CEN_ARS
Sequence file: RDB18572_B0J5g.seq check
>D07393A6_B0J5_2_Amp_R_H01_08_ABI08.ab1
    1 NNNNNNNNNN NNNNNNNNNA NCANTTATNN GGGTTATTGT CTCNTGAGCG GANACATATT
   61 TGAATGTATT TAGAAAAATA AACAAATAGG GGTTCCGCGC ACATTTCCCC GAAAAGTGCC
  121 ACCTGGGTCC TTTTCATCAC GTGCTATAAA AATAATTATA ATTTAAATTT TTTAATATAA
  181 ATATATAAAT TAAAAATAGA AAGTAAAAAA AGAAATTAAA GAAAAAATAG TTTTTGTTTT
  241 CCGAAGATGT AAAAGACTCT AGGGGGATCG CCAACAAATA CTACCTTTTA TCTTGCTCTT
  301 CCTGCTCTCA GGTATTAATG CCGAATTGTT TCATCTTGTC TGTGTAGAAG ACCACACACG
  361 AAAATCCTGT GATTTTACAT TTTACTTATC GTTAATCGAA TGTATATCTA TTTAATCTGC
  421 TTTTCTTGTC TAATAAATAT ATATGTAAAG TACGCTTTTT GTTGAAATTT TTTAAACCTT
  481 TGTTTATTTT TTTTTCTTCA TTCCGTAACT CTTCTACCTT CTTTATTTAC TTTCTAAAAT
  541 CCAAATACAA AACATAAAAA TAAATAAACA CAGAGTAAAT TCCCAAATTA TTCCATCATT
  601 AAAAGATACG AGGCGCGTGT AAGTTACAGG CAAGCGATCC GTCTAAGAAA CCATTATTAT
  661 CATGACATTA ACCTATAAAA ATAGGCGTAT CACGAGGCCC TTTCGTCTCG CGCGTTTCGG
  721 TGANGACGGT GAAAACCTCT GACACATGCA GCTCCCGGAN ACGGTCACAG CTTGTCTGTA
  781 AGCGGANGCC GGGANCAGAN AAGCCCGTCA NGGCGCGNCA GCGGGTGTTG GCGGGNGTCG
  841 GGGCTGGCTT ANCTATGCGG CATCAGANCA NANTGTACTG ANAGTGNACC ATANCACCTT
  901 TTCNNTTCAT CATTTTTTTT TTANNCTTTT TTTTGANTNN NNNTNNNNTN NAAATTTTTT
961 TGNNTCNGNN NNCNNNNNNN NNN
//

Please visit Sequencing and PCR primers for primer information.


References

Original, user report and related articles

original Kakimoto, Y., Visualizing multiple inter-organelle contact sites using the organelle-targeted split-GFP system. Sci. Rep. 8 (1): 6175 (2018). PMID 29670150.

2020.11.06

GNP_filter3_RDBDEP_html_201104.pl