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pSFL68 pRS314-GPDp-Erg6-V5-GFP11

Expression vector of S. cerevisiae sterol 24-C-methyltransferase (Erg6) fused with V5-GFP11 at C-terminus.

Catalog number RDB18574
Resource name pSFL68 pRS314-GPDp-Erg6-V5-GFP11
Clone info. The split-GFP system clone for visualizing organelle contact sites in vivo. Expression vector of S. cerevisiae sterol 24-C-methyltransferase (Erg6) fused with V5-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 pRS314 (plasmid)
Selectable markers Ampicillin (E. coli), TRP1 (S. cerevisiae).
Gene/insert name S. cerevisiae ERG6 cDNA Aequorea victoria GFP 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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Schedule A (GFP-A) [Link]
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RDB18574 pSFL68 pRS314-GPDp-Erg6-V5-GFP11 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 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 RDB18574_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)
Sequence file: RDB18574_B0J5a.seq check
>D07393A8_B0J5_3_HB-NOS_R_A03_03_ABI24.ab1
    1 NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNGNAACGCC NGGGTTTTCC CAGTCACGAC
   61 GTTGTAAAAC GACGGCCAGT GAATTGTAAT ACGACTCACT ATAGGGCGAA TTGGAGCTCA
  121 GTTTATCATT ATCAATACTC GCCATTTCAA AGAATACGTA AATAATTAAT AGTAGTGATT
  181 TTCCTAACTT TATTTAGTCA AAAAATTAGC CTTTTAATTC TGCTGTAACC CGTACATGCC
  241 CAAAATAGGG GGCGGGTTAC ACAGAATATA TAACATCGTA GGTGTCTGGG TGAACAGTTT
  301 ATTCCTGGCA TCCACTAAAT ATAATGGAGC CCGCTTTTTA AGCTGGCATC CAGAAAAAAA
  361 AAGAATCCCA GCACCAAAAT ATTGTTTTCT TCACCAACCA TCAGTTCATA GGTCCATTCT
  421 CTTAGCGCAA CTACAGAGAA CAGGGGCACA AACAGGCAAA AAACGGGCAC AACCTCAATG
  481 GAGTGATGCA ACCTGCCTGG AGTAAATGAT GACACAAGGC AATTGACCCA CGCATGTATC
  541 TATCTCATTT TCTTACACCT TCTATTACCT TCTGCTCTCT CTGATTTGGA AAAAGCTGAA
  601 AAAAAAGGTT GAAACCAGTT CCCTGAAATT ATTCCCCTAC TTGACTAATA AGTATATAAA
  661 GACGGTAGGT ATTGATTGTA ATTCTGTAAA TCTATTTCTT AAACTTCTTA AATTCTACTT
  721 TTATAGTTAG TCTTTTTTTT AGTTTTAAAA CACCAGAACT TAGTTTCGAC NGANGCGGCC
  781 GCNTGANTGA AACAGAATTG AGAAAAAGAC AGGNCCNATT CACTANGGAG TTACATGGTG
  841 ATGATATTGG TAAAAAGANN GGNTTGANNG CATTGATGTC GAANNAACAA CTCTGCNCNA
  901 ANNNNANCCG TNNNNAANTA CTTGANAAAT TGGGNNGGNA GAANCGATNA ANANGCCNNA
  961 NAANNNCNTC NTGNNNNANT ATANNGNNNN CNCNCNNTNC CTACNNNNAN NNNNTTNNNN
1021 ANNNNNNNNN N
//
Primer: SV40pA3'Element Reverse
Region: T3 pro, insert 3' (CYC1 ter, V5-GFP11 3')
Sequence file: RDB18574_B0J5b.seq check
>D07393A8_B0J5_3_SV40A3'ElementReverse_B03_06_ABI24.ab1
    1 NNNNNNNNNN NNNNNNNNNN NNNNNNNNGN NNAANNCNNG CAACGCGGCC TTTTTACGGT
   61 TCCTGGCCTT TTGCTGGCCT TTTGCTCACA TGTTCTTTCC TGCGTTATCC CCTGATTCTG
  121 TGGATAACCG TATTACCGCC TTTGAGTGAG CTGATACCGC TCGCCGCAGC CGAACGACCG
  181 AGCGCAGCGA GTCAGTGAGC GAGGAAGCGG AAGAGCGCCC AATACGCAAA CCGCCTCTCC
  241 CCGCGCGTTG GCCGATTCAT TAATGCAGCT GGCACGACAG GTTTCCCGAC TGGAAAGCGG
  301 GCAGTGAGCG CAACGCAATT AATGTGAGTT AGCTCACTCA TTAGGCACCC CAGGCTTTAC
  361 ACTTTATGCT TCCGGCTCGT ATGTTGTGTG GAATTGTGAG CGGATAACAA TTTCACACAG
  421 GAAACAGCTA TGACCATGAT TACGCCAAGC TCGAAATTAA CCCTCACTAA AGGGAACAAA
  481 AGCTGGTACC GGCCGCAAAT TAAAGCCTTC GAGCGTCCCA AAACCTTCTC AAGCAAGGTT
  541 TTCAGTATAA TGTTACATGC GTACACGCGT CTGTACAGAA AAAAAAGAAA AATTTGAAAT
  601 ATAAATAACG TTCTTAATAC TAACATAACT ATAAAAAAAT AAATAGGGAC CTAGACTTCA
  661 GGTTGTCTAA CTCCTTCCTT TTCGGTTAGA GCGGATGTGG GGGGAGGGCG TGAATGTAAG
  721 CGTGACATAA CTAATTACAT GACTCGAGGT CGACGGTATC GATAAGCTTG ATATCGAATT
  781 CTTANGTAAT GCCTGCAGCA TTGACATATT CGTGTAGAAN CATATGATCT CTGCTTGTGG
  841 AACCNCCACC ACTTCCNCCA GTTGAGTCTA AGCCCNAAAN ANGGTTNNGT ATGGGTTTCC
  901 CANAANCCTC CGGCNNANGN NCCTNGANTT GCTNNTTGGG AANTTTGGGA NGGGGNTNCG
  961 GNGTTTTCTG GNNTNNNNAN CNNNNAAAAG CATCATNNGN NNNGNNNNAN NTTNGNNNNN
1021 NNNNN
//
Primer: TRP1pro_F
Region: TRP1
Sequence file: RDB18574_B0J5c.seq check
>D07393A8_B0J5_3_TRP1pro_F_C03_09_ABI24.ab1
    1 NNNNNNNNNN NNNNNNNNNN NNAGGCAGCT TGGANTANGT CTGTTATTAA TTTCNCAGGT
   61 AGTTCTGGTC CNTTGGTGAA AGTTTGCGGC TTGCAGAGCA CAGAGGCCGC AGAATGTGCT
  121 CTAGATTCCG ATGCTGACTT GCTGGGTATT ATATGTGTGC CCAATAGAAA GAGAACAATT
  181 GACCCGGTTA TTGCAAGGAA AATTTCAAGT CTTGTAAAAG CATATAAAAA TAGTTCAGGC
  241 ACTCCGAAAT ACTTGGTTGG CGTGTTTCGT AATCAACCTA AGGAGGATGT TTTGGCTCTG
  301 GTCAATGATT ACGGCATTGA TATCGTCCAA CTGCATGGAG ATGAGTCGTG GCAAGAATAC
  361 CAAGAGTTCC TCGGTTTGCC AGTTATTAAA AGACTCGTAT TTCCAAAAGA CTGCAACATA
  421 CTACTCAGTG CAGCTTCACA GAAACCTCAT TCGTTTATTC CCTTGTTTGA TTCAGAAGCA
  481 GGTGGGACAG GTGAACTTTT GGATTGGAAC TCGATTTCTG ACTGGGTTGG AAGGCAAGAG
  541 AGCCCCGAAA GCTTACATTT TATGTTAGCT GGTGGACTGA CGCCAGAAAA TGTTGGTGAT
  601 GCGCTTAGAT TAAATGGCGT TATTGGTGTT GATGTAAGCG GAGGTGTGGA GACAAATGGT
  661 GTAAAAGACT CTAACAAAAT AGCAAATTTC GTCAAAAATG CTAAGAAATA GGTTATTACT
  721 GAGTAGTATT TATTTAAGTA TTGTTTGTGC ACTTGCCGAT CTATGCGGTG TGAAATACCG
  781 CACAGATGCG TAAGGAGAAA ATACCGCATC ANGAAATTGT AAGCGTTAAT ATTTTGTTAA
  841 AATTCGCGTT AAATTTTTGT NNAATCAGCT CATTTTTTAA CCAATANGCC GAAATCGGCA
  901 AAATCCCTTA TAAATCNAAA GAATANANCG ANATAGGGTT GANTGNTGNT CCNGTTTGGN
  961 ANNAGAGTNN NCTATTAAAG AACGNGNNNN NNNNCNNNNA NNNNNAAAAA NCGNCTATCN
1021 NNNNNNNGNN NNNNNNNN
//
Primer: CYC1ter_R
Region: insert mid (CYC1 ter, V5-GFP11, Erg6 3')
Sequence file: RDB18574_B0J5d.seq check
>D07393A8_B0J5_3_CYC1ter_R_F03_18_ABI24.ab1
    1 NNNNNNNNNN NNNNNNNNNN GCGGNNGNGG GGGGAGGGCG TGAATGTAAG CGTGACATAA
   61 CTAATTACAT GACTCGAGGT CGACGGTATC GATAAGCTTG ATATCGAATT CTTAGGTAAT
  121 GCCTGCAGCA TTGACATATT CGTGTAGAAC CATATGATCT CTGCTTGTGG AACCACCACC
  181 ACTTCCACCA GTTGAGTCTA AGCCCAAAAG AGGGTTTGGT ATGGGTTTCC CAGAACCTCC
  241 GGCAGAGGAT CCTTGAGTTG CTTCTTGGGA AGTTTGGGAG GGGGTTTCGG CGTTTTCTGG
  301 CTTCCTAGCG ACGAAAAGCA TCATTGGAGT GAATAACTTG GACTTACCAC CGGCAACTAA
  361 ACCAACCGCA GCATTTTCTA GAGCAGCAGT AACTTCCTTG GAACCTTCTG GGGCTAGACC
  421 TAATTTTTCC ATTACAGTAA CCATTGCTGT AGTAAATTGT CTACCCAAGT AAGAAGTTCT
  481 GAAAAATGTG GCCAAATTAG CTAAGTTTTG AACGTACTTC CACTCACCAG TTAATGGGTA
  541 ATACCAAGGG ATTTCATCAT CATTGTCCGC CAGGTCTTCG CTAACGAGGA CTTCGAAACC
  601 ACAGTTCTTC AATGCTTTCC TAGCCACGTC GACATGGAAC ATCTTTGGGA TACCATCACC
  661 TAGTTCAATT TCATAAGCGA TCTTTCTATG TTCAGGATTG TTTTCGTCAT ATTTATCAGT
  721 CATTACCCAT TCGTAAACAG CAAAGGTANC ACCCGGTTTC AAAACCTTGT AGATTTCGCT
  781 GTATACACCT TCTAATTTTG GAGCGTGACA TGTGGCCTCA ATTGCATAAA CTTTGTCNAA
  841 AGTGTTTTCT TCGAAATCCA TTTTCNTGAA ATCACCCTTT ACAAAGTCCA TTTGGTCANT
  901 CAAATTGTAT TTTTTTANCG NAATATTTTG NCNNNNCAAT TTGGNAATCG TTATTGNTTA
  961 NACNNATGAN GTTNCANNCN GNNAATNNNG NNNTCTNNNT NGNTGGGNCC NNNNCACNNN
1021 NNNNNNN
//
Primer: TDH3pro_F
Region: insert mid (GPD pro, Erg6 5')
Sequence file: RDB18574_B0J5e.seq check
>D07393A8_B0J5_3_TDH3pro_F_D03_12_ABI24.ab1
    1 NNNNNNNNNN NNNNNNNNNN NTGNNNTTCN GNAAATCTAT TTCTTAAACT TCTTAAATTC
   61 TACTTTTATA GTTAGTCTNT NNTTTAGTTT TAAAACACCA GAACTTAGTT TCGACGGATG
  121 CGGCCGCATG AGTGAAACAG AATTGAGAAA AAGACAGGCC CAATTCACTA GGGAGTTACA
  181 TGGTGATGAT ATTGGTAAAA AGACAGGTTT GAGTGCATTG ATGTCGAAGA ACAACTCTGC
  241 CCAAAAGGAA GCCGTTCAGA AGTACTTGAG AAATTGGGAT GGTAGAACCG ATAAAGATGC
  301 CGAAGAACGT CGTCTTGAGG ATTATAATGA AGCCACACAT TCCTACTATA ACGTCGTTAC
  361 AGATTTCTAT GAATATGGTT GGGGTTCCTC TTTCCATTTC AGCAGATTTT ATAAAGGTGA
  421 GAGTTTCGCT GCCTCGATAG CAAGACATGA ACATTATTTA GCTTACAAGG CTGGTATTCA
  481 AAGAGGCGAT TTAGTTCTCG ACGTTGGTTG TGGTGTTGGG GGCCCAGCAA GAGAGATTGC
  541 AAGATTTACC GGTTGTAACG TCATCGGTCT AAACAATAAC GATTACCAAA TTGCCAAGGC
  601 AAAATATTAC GCTAAAAAAT ACAATTTGAG TGACCAAATG GACTTTGTAA AGGGTGATTT
  661 CATGAAAATG GATTTCGAAG AAAACACTTT CGACAAAGTT TATGCAATTG AGGCCACATG
  721 TCACGCTCCA AAATTAGAAG GTGTATACAG CGAAATCTAC AAGGTTTTGA AACCGGGNGG
  781 TACCTTTGCT GTTTACGANT GGGTAATGAC TGATAAATAT GACGAAAACA ATCCTGAACA
  841 TAGAAAGATC GCTTATGAAA TTGANCTANG TGANGGTATC CCNAAGANGT TNCANGTCNA
  901 CGNGGCTAGG AAAGCATTGA AGAANTGTGN TTTCNAAGTC CTCGTTAGCG ANNNNNNNNN
961 N
//
Primer: Amp_R
Region: AmpR pro, CEN_ARS
Sequence file: RDB18574_B0J5f.seq check
>D07393A8_B0J5_3_Amp_R_E03_15_ABI24.ab1
    1 NNNNNNNNNN NNNNNNNNNN NNATTTATCN GGGTTATTGT CTCATGAGCG GATACATATT
   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 TGATGACGGT GAAAACCTCT GACACATGCA GCTCCCGGAG ACGGTCACAG CTTGTCTGTA
  781 AGCGGATGCC GGGAGCAGAC AAGCCCGTCA GGGCGCGTCA GCGGGTGTTG GCGGGTGTCG
  841 GGGCTGGCTT AACTATGCGG CATCNNANCA GANTGNACTG AGANNGNNCC NTANATCACG
  901 ANATTACTAT ATATATAATA TAGGAAGCAT TTAATANAAN NGCATCGTAA TATATGTGTA
  961 CTTTGCAGTT ATGANNNNNN ANGNNANNAN NGNNNNANAN NCNTTANNGN AAAATANNNN
1021 GNNNCNNTNN NNNNNNNN
//

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.

2021.01.21

GNP_filter3_RDBDEP_html_201222.pl