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pYC135 pBS-GPDp-Ifa38-V5-GFP11-CyC1ter-natNT2

The split-GFP system clone for visualizing organelle contact sites in S. cerevisiae. probe: V5-GFP11; location: endoplasmic reticulum.

Catalog number RDB18656
Resource name pYC135 pBS-GPDp-Ifa38-V5-GFP11-CyC1ter-natNT2
Alternative name pYC135, pBS-GPDp-Ifa38-V5-GFP11-CyC1ter-natNT2
Clone info. The split-GFP system clone for visualizing organelle contact sites in vivo. Expression vector of S. cerevisiae ketoreductase (Ifa38) fused with V5-GFP11 at C-terminus, GPD promoter, CYC1 terminator, natNT2 (clonNAT resistant marker).
Vector backbone pBlueScript SK- (plasmid)
Size of vector backbone 3.0 kb
Selectable markers Ampicillin (E. coli), Nourseothricin (yeast).
Gene/insert name S. cerevisiae IFA38 cDNA Aequorea victoria GFP cDNA
Depositor|Developer Tamura, Yasushi |
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External Database
S. cerevisiae IFA38 Aequorea victoria GFP

          Reference sequence
            
           
          Sequence (full) RDB18656zzt01.seq compiled by us

          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.(Tashiro, S. et al. Front. Cell Dev. Biol. 8(1): 6175, 2020)
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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(Tashiro, S. et al. Front. Cell Dev. Biol. 8(1): 6175, 2020)
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          RDB18656 pYC135 pBS-GPDp-Ifa38-V5-GFP11-CyC1ter-natNT2 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 pYC135 pBS-GPDp-Ifa38-V5-GFP11-CyC1ter-natNT2 was provided by the RIKEN BRC through the National BioResource Project of the MEXT, Japan (cat. RDB18656).

          Reference section:

          Tashiro, S., Kakimoto, Y., Shinmyo, M., Fujimoto, S., Tamura, Y., Improved Split-GFP Systems for Visualizing Organelle Contact Sites in Yeast and Human Cells. Front. Cell Dev. Biol. 8: 571388 (2020). PMID 33330450.

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


          References and tips

          Featured content

          Featured content Visualizing multiple inter-organelle contact sites (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 RDB18656_B0Lnp1-1.pdf check

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

          Primer: f1ori_F2 (Pr0308)
          Region: f1 ori, T7 pro, insert 5'(GPD pro)
          Sequence file: RDB18656_B0Lna.seq check
          >D07536A2_B0Ln_1_f1ori_F2_A10_01_ABI24.ab1
              1 NNNNNNNNNN NNNNGNNNTT TNNCGANTTC GNNCTATTGG TTAAAAAAAT GAGCTGATTT
             61 AACAAAAATT TAACGCGAAT TTTAACAAAA TATTAACGCT TACAATTTCC ATTCGCCATT
            121 CAGGCTGCGC AACTGTTGGG AAGGGCGATC GGTGCGGGCC TCTTCGCTAT TACGCCAGCT
            181 GGCGAAAGGG GGATGTGCTG CAAGGCGATT AAGTTGGGTA ACGCCAGGGT TTTCCCAGTC
            241 ACGACGTTGT AAAACGACGG CCAGTGAGCG CGCGTAATAC GACTCACTAT AGGGCGAATT
            301 GGGTACCGGG CCCCCCCTCG AGGTCGACGG TATCGATAAG CTTGATATCG AATTAGTTTA
            361 TCATTATCAA TACTCGCCAT TTCAAAGAAT ACGTAAATAA TTAATAGTAG TGATTTTCCT
            421 AACTTTATTT AGTCAAAAAA TTAGCCTTTT AATTCTGCTG TAACCCGTAC ATGCCCAAAA
            481 TAGGGGGCGG GTTACACAGA ATATATAACA TCGTAGGTGT CTGGGTGAAC AGTTTATTCC
            541 TGGCATCCAC TAAATATAAT GGAGCCCGCT TTTTAAGCTG GCATCCAGAA AAAAAAAGAA
            601 TCCCAGCACC AAAATATTGT TTTCTTCACC AACCATCAGT TCATAGGTCC ATTCTCTTAG
            661 CGCAACTACA GAGAACAGGG GCACAAACAG GCAAAAAACG GGCACAACCT CAATGGAGTG
            721 ATGCAACCTG CCTGGAGTAA ATGATGACAC AAGGCAATTG ACCCACGCAT GTATCTATCT
            781 CATTTTCTTA CACCTTCTAT TACCTTCTGC TCTCTCTGAT TTGGAAAAAG CTGAAAAAAA
            841 AGGTTGAAAC CAGTTCCCTG AAATTATTCC CCTACTTGAC TAATAAGTAT ATAAAGANGG
            901 NAGGTATTGA TTGNAATTCT GTAAATCTAT TTCNNAAACT NCTNNAATNN NACTTTTANA
            961 GTNAGTCTTT TTTTTTANTT TTAAAACACN NNAACTTNNT TNNNNNNNAN GNNGNNNNNN
          1021 NNNN
          //
          Primer: pUC-CAPbs_up_F2 (Pr0739)
          Region: T3 pro, insert 3'(TEF pro, natNT2 5')
          Sequence file: RDB18656_B0Lnb.seq check
          >D07536A2_B0Ln_1_pUC-CAPbs_up_F2_B10_04_ABI24.ab1
              1 NNNNNNNNNN NNNNNNNNNN NNNNNNNTNN NNCTTTATGC TTCCGGCTCG TATGTTGTGT
             61 GGAATTGTGA GCGGATAACA ATTTCACACA GGAAACAGCT ATGACCATGA TTACGCCAAG
            121 CGCGCAATTA ACCCTCACTA AAGGGAACAA AAGCTGGAGC TCCACCGCGG TGGCGGCCGC
            181 TCTAGAACTA GTGGATCCCG GGTTAATTAA GGCGCGCCAG ATCTGTTTAG CTTGCCTCGT
            241 CCCCGCCGGG TCACCCGGCC AGCGACATGG AGGCCCAGAA TACCCTCCTT GACAGTCTTG
            301 ACGTGCGCAG CTCAGGGGCA TGATGTGACT GTCGCCCGTA CATTTAGCCC ATACATCCCC
            361 ATGTATAATC ATTTGCATCC ATACATTTTG ATGGCCGCAC GGCGCGAAGC AAAAATTACG
            421 GCTCCTCGCT GCAGACCTGC GAGCAGGGAA ACGCTCCCCT CACAGACGCG TTGAATTGTC
            481 CCCACGCCGC GCCCCTGTAG AGAAATATAA AAGGTTAGGA TTTGCCACTG AGGTTCTTCT
            541 TTCATATACT TCCTTTTAAA ATCTTGCTAG GATACAGTTC TCACATCACA TCCGAACATA
            601 AACAACCATG GGTACCACTC TTGACGACAC GGCTTACCGG TACCGCACCA GTGTCCCGGG
            661 GGACGCCGAG GCCATCGAGG CACTGGATGG GTCCTTCACC ACCGACACCG TCTTCCGCGT
            721 CACCGCCACC GGGGACGGCT TCACCCTGCG GGANGTGCCG GTGGACCCGC CCCTGACCAA
            781 NGTGTTCCCC GACGACNAAT CGGACGANGA ATCGGANGAN GGGGAGGANG GCGACCCGGA
            841 NTCNCGGANG TTCGTCGCGT ACGGGGANNA NGGCGANCTG GCGGGCTTCG TGGNCGTCTC
            901 GNACTCCGGC NNGAANCGCC NGCTGANCGN CNANGANNTC GANGNCGCCC CGGANCACCG
            961 GGGGCNCGGG NNNGGGNGCG CGNNGANGGG NNNNGCGANN NANNNNGNCN NCNANNNGGN
          1021 NNNNNGNNNN NNNNNGNNNN NNNNNN
          //
          Primer: TDH3pro_F (Pr0431)
          Region: insert mid(GPD pro, Ifa38 5')
          Sequence file: RDB18656_B0Lnc.seq check
          >D07536A2_B0Ln_1_TDH3pro_F_C10_07_ABI24.ab1
              1 NNNNNNNNNN NNNNNTGNNT GNNNTTCTNN NNNNTATTTC TTAAACTTCT TAAATTCTAC
             61 TTTTATAGTT AGTCTTTTTT TTAGTTTTAA AACACCAGAA CTTAGTTTCG ACGGATGCGG
            121 CCGCATGACT TTTATGCAAC AGCTTCAAGA GGCTGGGGAA AGATTTAGGT GTATCAATGG
            181 TCTTTTATGG GTTGTTTTTG GGCTTGGTGT CTTAAAATGT ACAACGTTAT CACTTAGATT
            241 TTTAGCTCTT ATTTTTGATC TTTTTTTACT ACCAGCAGTC AATTTCGACA AGTATGGTGC
            301 TAAAACTGGT AAATACTGTG CTATCACTGG TGCAAGTGAC GGTATTGGTA AAGAATTTGC
            361 TAGACAAATG GCGAAACGTG GCTTCAACTT GGTATTGATC TCGAGAACGC AATCTAAATT
            421 GGAGGCTTTA CAAAAAGAAC TAGAAGATCA ACATCATGTC GTTGTAAAGA TTCTAGCCAT
            481 TGACATTGCG GAGGATAAAG AATCCAACTA TGAATCCATT AAGGAATTGT GTGCGCAGTT
            541 ACCAATCACC GTTTTGGTCA ATAATGTTGG TCAATCACAC TCCATTCCTG TTCCATTTTT
            601 GGAAACAGAA GAGAAGGAGC TTAGAAATAT TATCACTATC AATAACACTG CTACATTATT
            661 AATTACACAA ATCATTGCAC CAAAGATTGT GGAAACTGTG AAAGCTGAAA ACAAGAAGTC
            721 GGGTACTCGT GGTTTGATCT TAACCATGGG CTCATTTGGT GGTCTGATTC CCACCCCACT
            781 TTTGGCTACA TACAGTGGTT CGAAATCATT CTTACAAGGT TGGTCTAACT CTTTGGCTGG
            841 GGANTTATCT AAAGATGCTA TCGATGTTGA ATTAATCATT TCATATTNGG TCANTAGCTC
            901 GATGTCTAAA ATCAGAAGAN CATNTTTAAT GATCCNNANT CCNCANCAGT TTGTAAAATC
            961 CNCTTNNNNN NNNGNNNGNN GNNGCNNCNN NNNNCNNNAA ANNNNNNCTN NNNNNNNCNN
          1021 NTNNNNNGNN NNNNNNNN
          //
          Primer: CYC1ter_R (Pr0022)
          Region: insert mid(CYC1 ter, GFP11, V5 tag, Ifa38 3')
          Sequence file: RDB18656_B0Lnd.seq check
          >D07536A2_B0Ln_1_CYC1ter_R_D10_10_ABI24.ab1
              1 NNNNNNNNNN NNNNNNNNNN NNNNCNNNNN NGGGGGGANG GCGTGAATGT AAGCGTGACA
             61 TAACTAATTA CATGACTCGA GGTCGACGGT ATCGATAAGC TTGATATCGA ATTCTTAGGT
            121 AATGCCTGCA GCATTGACAT ATTCGTGTAG AACCATATGA TCTCTGCTTG TGGAACCACC
            181 ACCACTTCCA CCAGTTGAGT CTAAGCCCAA AAGAGGGTTT GGTATGGGTT TCCCAGAACC
            241 TCCGGCAGAG GATCCTTCCT TTTTAACCTG TCTTGCGGCT TTTTTTAAGG CTCTAATTCT
            301 GATAGATTTA TGGAAGGAAT AATTAATGGA GTTAACAATC TTAGAGTAAA CGCCAAAGGT
            361 CTCTGTAATT ACAAATTGAT ACACTGCGTG CGCCCAGTAA GGGGTCATAG TAGCGTATCT
            421 TTCTTGAGAG CCGCAGCGTC TGCCAACGCT TCTTAAAGTG GATTTTACAA ACTGTTGTGG
            481 ATTTGGGATC ATTAAAGATG ATCTTCTGAT TTTAGACATC GAGCTAGTGA CCAAATATGA
            541 AATGATTAAT TCAACATCGA TAGCATCTTT AGATAATTCC CCAGCCAAAG AGTTAGACCA
            601 ACCTTGTAAG AATGATTTCG AACCACTGTA TGTAGCCAAA AGTGGGGTGG GAATCAGACC
            661 ACCAAATGAG CCCATGGTTA AGATCAAACC ACGAGTACCC GACTTCTTGT TTTCAGCTTT
            721 CACAGTTTCC ACAATCTTTG GTGCAATGAT TTGTGTAATT AATAATGTAG CAGTGTTATT
            781 GATAGTGATA ATATTTCTAA GCTCCTTCTC TTCTGTTTCC NAAAATGGAA CAGGAATGGA
            841 GTGTGANTGA NCANCATTAT TGACCAAACG GTGATTGGTA ACTGCGCACA CAATTCCNTT
            901 AANGGANTCN TAGTTNGGAN NNTTTATCCT CCGCANTGTC NANGGNTANA ATCTTNNNNN
            961 NGACNTGATG TTGNNCTTCT ANTNCNTTTT NGNAAANNCN NNNNNTTTAN ANNGNNTNNN
          1021 NNANNNNNNT ANNNANNNNN NNNNN
          //
          Primer: CYC1ter_F (Pr0273)
          Region: insert mid(ADH1 ter, natNT2 3')
          Sequence file: RDB18656_B0Lne.seq check
          >D07536A2_B0Ln_1_CYC1ter_F_E10_13_ABI24.ab1
              1 NNNNNNNNNN NNNNNNNGGN NNNTNNNNNN NTTTAATTTG CGGCCGANCT CGATTACAAC
             61 AGGTGTTGTC CTCTGAGGAC ATAAAATACA CACCGAGATT CATCAACTCA TTGCTGGAGT
            121 TAGCATATCT ACAATTGGGT GAAATGGGGA GCGATTTGCA GGCATTTGCT CGGCATGCCG
            181 GTAGAGGTGT GGTCAATAAG AGCGACCTCA TGCTATACCT GAGAAAGCAA CCTGACCTAC
            241 AGGAAAGAGT TACTCAAGAA TAAGAATTTT CGTTTTAAAA CCTAAGAGTC ACTTTAAAAT
            301 TTGTATACAC TTATTTTTTT TATAACTTAT TTAATAATAA AAATCATAAA TCATAAGAAA
            361 TTCGCCTCGA GATTAGGGGC AGGGCATGCT CATGTAGAGC GCCTGCTCGC CGTCCGAGGC
            421 GGTGCCGTCG TACAGGGCGG TGTCCAGGCC GCAGAGGGTG AACCCCATCC GCCGGTACGC
            481 GTGGATCGCC GGTGCGTTGA CGTTGGTGAC CTCCAGCCAG AGGTGCCCGG CGCCCCGCTC
            541 GCGGGCGAAC TCCGTCGCGA GCCCCATCAA CGCGCGCCCG ACCCCGTGCC CCCGGTGCTC
            601 CGGGGCGACC TCGATGTCCT CGACGGTCAG CCGGCGGTTC CAGCCGGAGT ACGAGACGAC
            661 CACGAAGCCC GCCAGGTCGC CGTCGTCCCC GTACGCGACG AACGTCCGGG AGTCCGGGTC
            721 GCCGTCCTCC CCGTCGTCCG ATTCGTCGTC CGATTCGTCG TCNGGGAACA CCTTGGTCAG
            781 GGGCGGGTCC ACCGGCACCT CCCGCANGGT GAAGCCGTCC CCGGTGGNGG TGANGCGGAN
            841 NANGGTGTCG GNGGTGAANG ANCCATCCNG TGNCTCNATG GNCTCNGCGT CCCCCGGGAN
            901 ACTGGTGCGG TACCGGTAAG CCGNGNCGTN NNGANTGGTA CCNANNGNNN GNTTANNNNC
          961 GNNNGNGATG TGANANNNGN ANNCNTANCN NGANTTTNAA NGNNNNNANN NNNNNNNN
          //

          Please visit Sequencing and PCR primers for primer information.


          References

          Original, user report and related articles

          original Tashiro, S., Improved Split-GFP Systems for Visualizing Organelle Contact Sites in Yeast and Human Cells. Front. Cell Dev. Biol. 8: 571388 (2020). PMID 33330450.

          2023.05.01

          GNP_filter3_RDBDEP_html_230501.pl