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pYC137 pBS-GPDp-DPP1-V5-GFP11-CyC1ter-natNT2

The split-GFP system clone for visualizing organelle contact sites in vivo. Expression vector of S. cerevisiae DPP1 fused with V5-GFP11 at C-terminus.

Catalog number RDB18658
Resource name pYC137 pBS-GPDp-DPP1-V5-GFP11-CyC1ter-natNT2
Clone info. The split-GFP system clone for visualizing organelle contact sites in vivo. Expression vector of S. cerevisiae bifunctional diacylglycerol diphosphate phosphatase/phosphatidate phosphatase (DPP1) 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 DPP1 cDNA Aequorea victoria GFP cDNA
Depositor|Developer Tamura, Yasushi |
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External Database
S. cerevisiae DPP1 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 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.(Tashiro, S. et al. Front. Cell Dev. Biol. 8(1): 6175, 2020)
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          Schedule A (GFP-A) [Link]
          Remarks
          Please use Schedule A (GFP-A) additionally.
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          RDB18658 pYC137 pBS-GPDp-DPP1-V5-GFP11-CyC1ter-natNT2 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

          Sequence (full) RDB18658zzt01.seq compiled by us

          Original reference

          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.

          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 RDB18658_B0Lnp1-1.pdf check

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

          Primer: f1ori_F2
          Region: f1 ori, T7 pro, GPD pro
          Sequence file: RDB18658_B0Lna.seq check
          >D07536A4_B0Ln_2_f1ori_F2_F05_17_ABI24.ab1
              1 NNNNNNNNNN NNNNNNNNNT NNCGANTTCG NNCTATTGGT TAAAAAAATG AGCTGATTTA
             61 ACAAAAATTT AACGCGAATT TTAACAAAAT ATTAACGCTT ACAATTTCCA TTCGCCATTC
            121 AGGCTGCGCA ACTGTTGGGA AGGGCGATCG GTGCGGGCCT CTTCGCTATT ACGCCAGCTG
            181 GCGAAAGGGG GATGTGCTGC AAGGCGATTA AGTTGGGTAA CGCCAGGGTT TTCCCAGTCA
            241 CGACGTTGTA AAACGACGGC CAGTGAGCGC GCGTAATACG ACTCACTATA GGGCGAATTG
            301 GGTACCGGGC CCCCCCTCGA GGTCGACGGT ATTGATAAGC TTGATATCGA ATTAGTTTAT
            361 CATTATCAAT ACTCGCCATT TCAAAGAATA CGTAAATAAT TAATAGTAGT GATTTTCCTA
            421 ACTTTATTTA GTCAAAAAAT TAGCCTTTTA ATTCTGCTGT AACCCGTACA TGCCCAAAAT
            481 AGGGGGCGGG TTACACAGAA TATATAACAT CGTAGGTGTC TGGGTGAACA GTTTATTCCT
            541 GGCATCCACT AAATATAATG GAGCCCGCTT TTTAAGCTGG CATCCAGAAA AAAAAAGAAT
            601 CCCAGCACCA AAATATTGTT TTCTTCACCA ACCATCAGTT CATAGGTCCA TTCTCTTAGC
            661 GCAACTACAG AGAACAGGGG CACAAACAGG CAAAAAACGG GCACAACCTC AATGGAGTGA
            721 TGCAACCTGC CTGGAGTAAA TGATGACACA AGGCAATTGA CCCACGCATG TATCTATCTC
            781 ATTTTCTTAC ACCTTCTATT ACCTTCTGCT CTCTCTGATT TGGAAAAAGC TGAAAAAAAA
            841 GGTTGAAACC AGTTCCCTGA AATTATTCCN CTACTTGANT AATANNNATA TNAAGANNGN
            901 AGGNANNGAT TGNNATTCTG TAAATCTATT TCTTAAACTT CTNAAATTCT ACTTTTATAG
            961 TTAGNCTTTT TTTTNANTTT TAAAANACNN NNNNNTNNNT TNCNANNNAN GCNGNNNNCN
           1021 TNNNNNGNNN TTNCNNNTNN NTNNAANNNN NNTTNNNNNN NNNGNNNNNA NNNNNNNNNN
          //
          Primer: pUC-CAPbs_up_F2
          Region: T3 pro, natNT2 (TEF pro, NrsR)
          Sequence file: RDB18658_B0Lnb.seq check
          >D07536A4_B0Ln_2_pUC-CAPbs_up_F2_G05_20_ABI24.ab1
              1 NNNNNNNNNN NNNNNNNNNN NNNNNGCTTN NNNCTTNATG CTTCCGGCTC GTATGTTGTG
             61 TGGAATTGTG AGCGGATAAC AATTTCACAC AGGAAACAGC TATGACCATG ATTACGCCAA
            121 GCGCGCAATT AACCCTCACT AAAGGGAACA AAAGCTGGAG CTCCACCGCG GTGGCGGCCG
            181 CTCTAGAACT AGTGGATCCC GGGTTAATTA AGGCGCGCCA GATCTGTTTA GCTTGCCTCG
            241 TCCCCGCCGG GTCACCCGGC CAGCGACATG GAGGCCCAGA ATACCCTCCT TGACAGTCTT
            301 GACGTGCGCA GCTCAGGGGC ATGATGTGAC TGTCGCCCGT ACATTTAGCC CATACATCCC
            361 CATGTATAAT CATTTGCATC CATACATTTT GATGGCCGCA CGGCGCGAAG CAAAAATTAC
            421 GGCTCCTCGC TGCAGACCTG CGAGCAGGGA AACGCTCCCC TCACAGACGC GTTGAATTGT
            481 CCCCACGCCG CGCCCCTGTA GAGAAATATA AAAGGTTAGG ATTTGCCACT GAGGTTCTTC
            541 TTTCATATAC TTCCTTTTAA AATCTTGCTA GGATACAGTT CTCACATCAC ATCCGAACAT
            601 AAACAACCAT GGGTACCACT CTTGACGACA CGGCTTACCG GTACCGCACC AGTGTCCCGG
            661 GGGACGCCGA GGCCATCGAG GCACTGGATG GGTCCTTCAC CACCGACACC GTCTTCCGCG
            721 TCACCGCCAC CGGGGACGGC TTCACCCTGC GGGAGGTGCC GGTGGANCCG CCCCTGANNA
            781 AGGTGTTCCC CGACGACGAA TCGGACGACG AATCGGANGA NGGGGAGGAN GGCGANCCGG
            841 ACTCCCGGAN GTTCGTCGCG NACGGGGNNN ANNGCGACCT NNNGGGCTTC GNGNNCGNCT
            901 CGNACTCCGG NNNGAANCGN CGGCTGANCG NCNANGANNT CNAGGTCGCC CCNNANCACC
            961 NGGGGGCNCG GGGNCNGGCG CGCGTNGANG GGGNNNNNGA NGNNNNNNGN NNGCNANNNG
          1021 GNNNNNNGNN NNCNNNGGNN NNANNNNNNN NNN
          //
          Primer: TDH3pro_F
          Region: GPD pro, DPP1 5'
          Sequence file: RDB18658_B0Lnc.seq check
          >D07536A4_B0Ln_2_TDH3pro_F_E05_14_ABI24.ab1
              1 NNNNNNNNNN NNNNNNNNNN TGNNTGNAAT TCTGNNNNNC TATTTCTTAA ACTTCTTAAA
             61 TTCTACTTTT ATAGTTAGTC TTTTTTTTAG TTTTAAAACA CCAGAACTTA GTTTCGACGG
            121 ATGCGGCCGC ATGAACAGAG TTTCGTTTAT TAAAACGCCT TTCAACATAG GGGCGAAATG
            181 GAGATTAGAA GATGTCTTTT TGCTCATTAT CATGATACTT CTTAACTACC CAGTGTATTA
            241 CCAACAACCG TTCGAACGTC AGTTTTACAT TAACGATCTC ACTATATCGC ATCCTTATGC
            301 GACAACTGAA CGTGTAAATA ACAACATGTT GTTTGTTTAT AGTTTTGTCG TGCCATCTTT
            361 AACCATATTG ATAATTGGTT CCATTTTGGC CGATAGAAGA CATTTGATTT TTATTTTGTA
            421 CACATCTCTC CTTGGTTTAT CACTCGCTTG GTTCAGTACG AGTTTCTTTA CAAACTTCAT
            481 CAAGAATTGG ATTGGAAGAC TAAGACCAGA TTTTCTAGAT CGTTGCCAAC CTGTTGAAGG
            541 CTTGCCATTG GACACTTTAT TTACTGCAAA AGATGTGTGT ACGACTAAGA ATCACGAACG
            601 TCTGTTGGAT GGGTTTAGGA CAACTCCGTC AGGTCATTCA AGTGAAAGCT TTGCAGGACT
            661 GGGTTATTTG TACTTCTGGC TATGTGGGCA ACTTTTGACT GAATCACCGT TGATGCCTTT
            721 ATGGAGAAAA ATGGTGGCCT TTCTACCACT GTTAGGAGCT GCACTAATTG CTCTATCCAG
            781 AACTCAAGAN TACAGACATC ATTTCGTCGA TGTAATTTTA GGNTCTATGT TGGGTTATAT
            841 AATGGCACAC TTTTTCTACA GAAGAATCTT NCCACCCATT GATGATCCTC TTNCGTTCNA
            901 ACNNTTGATG GANGATTCNN ANGNCACCCN NGNANGAAGC AGNCACCCNT CNNANGNNCC
          961 CNGNANGANG AANTNCATNC TTTGTCCNAN NNANNNNATG NNNNNNNNNN
          //
          Primer: CYC1ter_R
          Region: CYC1 ter, GFP11, V5 tag, DPP1 3'
          Sequence file: RDB18658_B0Lnd.seq check
          >D07536A4_B0Ln_2_CYC1ter_R_B12_06_ABI24.ab1
              1 NNNNNNNNNN NNNNNNNNNN NNNNNNNGTG GGGGGNGGGC GTGAATGTAA GCGTGACATA
             61 ACTAATTACA TGACTCGAGG TCGACGGTAT CGATAAGCTT GATATCGAAT TCTTAGGTAA
            121 TGCCTGCAGC ATTGACATAT TCGTGTAGAA CCATATGATC TCTGCTTGTG GAACCACCAC
            181 CACTTCCACC AGTTGAGTCT AAGCCCAAAA GAGGGTTTGG TATGGGTTTC CCAGAACCTC
            241 CGGCAGAGGA TCCACTAGTC ATACCTTCAT CGGACAAAGG ATGTAATTCC TCATCCGGGA
            301 TCCTCTGATG GGTGACTGCT TCCTCCAGGG TGACATCTGA ATCGTCCATC AATGGTTTGA
            361 ACGGAAGAGG ATCATCAATG GGTGGGAAGA TTCTTCTGTA GAAAAAGTGT GCCATTATAT
            421 AACCCAACAT AGACCCTAAA ATTACATCGA CGAAATGATG TCTGTAATCT TGAGTTCTGG
            481 ATAGAGCAAT TAGTGCAGCT CCTAACAGTG GTAGAAAGGC CACCATTTTT CTCCATAAAG
            541 GCATCAACGG TGATTCAGTC AAAAGTTGCC CACATAGCCA GAAGTACAAA TAACCCAGTC
            601 CTGCAAAGCT TTCACTTGAA TGACCTGACG GAGTTGTCCT AAACCCATCC AACAGACGTT
            661 CGTGATTCTT AGTCGTACAC ACATCTTTTG CAGTAAATAA AGTGTCCAAT GGCAAGCCTT
            721 CAACAGGTTG GCAACGATCT AGAAAATCTG GTCTTAGTCT TCCAATCCAA TTCTTGATGA
            781 AGTTTGTAAA GAAACTCGTA CTGAACCAAG CGAGTGATAA ACCAAGGAGA GATGTGTACA
            841 AAATAAAAAT CAAATGTCTT CTATCGGCCA AAATGGAACC NATTATCAAT ATGGTTAAAG
            901 ANGGNNCGAC AAAACTATAA ACAAACAACN TGTTGNTATT NNCACGTTCA GTTGNCGCAT
          961 AANGNTGCNA TATANTGAGN TCGNNNATGN AAAACTNNNN TTCNNNNNNN NNN
          //
          Primer: CYC1ter_F
          Region: CYC1 ter, natNT2 (ADH1 ter, NrsR)
          Sequence file: RDB18658_B0Lne.seq check
          >D07536A4_B0Ln_2_CYC1ter_F_D05_11_ABI24.ab1
              1 NNNNNNNNNN NTTNNNNNNN CTCGNNGGNT TTAATTTGCG GCCGAGCTCG ATTACAACAG
             61 GTGTTGTCCT CTGAGGACAT AAAATACACA CCGAGATTCA TCAACTCATT GCTGGAGTTA
            121 GCATATCTAC AATTGGGTGA AATGGGGAGC GATTTGCAGG CATTTGCTCG GCATGCCGGT
            181 AGAGGTGTGG TCAATAAGAG CGACCTCATG CTATACCTGA GAAAGCAACC TGACCTACAG
            241 GAAAGAGTTA CTCAAGAATA AGAATTTTCG TTTTAAAACC TAAGAGTCAC TTTAAAATTT
            301 GTATACACTT ATTTTTTTTA TAACTTATTT AATAATAAAA ATCATAAATC ATAAGAAATT
            361 CGCCTCGAGA TTAGGGGCAG GGCATGCTCA TGTAGAGCGC CTGCTCGCCG TCCGAGGCGG
            421 TGCCGTCGTA CAGGGCGGTG TCCAGGCCGC AGAGGGTGAA CCCCATCCGC CGGTACGCGT
            481 GGATCGCCGG TGCGTTGACG TTGGTGACCT CCAGCCAGAG GTGCCCGGCG CCCCGCTCGC
            541 GGGCGAACTC CGTCGCGAGC CCCATCAACG CGCGCCCGAC CCCGTGCCCC CGGTGCTCCG
            601 GGGCGACCTC GATGTCCTCG ACGGTCAGCC GGCGGTTCCA GCCGGAGTAC GAGACGACCA
            661 CGAAGCCCGC CAGGTCGCCG TCGTCCCCGT ACGCGACGAA CGTCCGGGAG TCCGGGTCGC
            721 CGTCCTCCCC GTCGTCCGAT TCGTCGTCCG ATTCGTCGTC GGGGAACACC TTGGTCAGGG
            781 GCGGGTCCAC CGGCACCTCC CGCANGGTGA AGCCGTCCCC GGTGGCGGTG ANGCGGAAGA
            841 CGGTGTCGGT GGTGAAGGAN CCATCCNGNG CCTCGATGGN NTCGGCGNCC CCCGGGANAC
            901 TGGTGCGGTA NCGGTAAGCC GNGNCNNCNA GANNGGTACC CATGGNTGNT TATGTNCNNN
            961 NGTGATGTGA GNNNTGTATC NTANCNNGAN TTNNANNNNN GNANNNNAAA NNNNNNCNNN
          1021 NNNNNNANNN NNNNNCNNTT TNNNNNTTNN NNNNNNNGGN NNNNNNNNN
          //

          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.

          2021.11.17

          GNP_filter3_RDBDEP_html_211111.pl