Westermann Lab Plasmids
mtGFP plasmids can conveniently be obtained from Addgene
Addgene: Benedikt Westermann
Reference
Westermann, B. and Neupert, W. (2000). Mitochondria-targeted green fluorescent proteins: convenient tools for the study of organelle biogenesis in Saccharomyces cerevisiae. Yeast 16: 1421-1427. PubMed.
Figure 1. Fusion of a mitochondrial presequence, pre Su9, to GFP. The amino acid sequence and the corresponding DNA coding region of the joint between the mitochondrial presequence and GFP are shown. The processing site for the mitochondrial matrix processing peptidase (MPP) is indicated. Relevant restriction sites in the linker region are shown.
Sequence information for the mtGFP plasmid series
Unfortunately, electronic sequence information for the pYX and pVT100U vector backbones is not available. Below is the complete sequence of pYES-mtGFP, and of the mtGFP cassette. Please note that in some constructs mtGFP was cloned HindIII/EcoRI, and in others EcoRI/HindIII. The mtBFP insert was digested with BamHI/BglII and religated (thereby the restriction sites shown in Fig.1 were removed).
Figure 2. Plasmids for expression of mitochondria-targeted GFP in yeast. Promoters are indicated by black arrows, 'pre' is the mitochondrial presequence, 'GFP' is the S65T allele, and 'BFP' is a blue-shifted version of GFP. Replication origins, transformation markers for yeast, and relevant restriction sites are indicated. Underlined restriction sites are known not to be unique; not all of the non-underlined sites have been checked. Plasmid maps are not drawn to scale.
- mtGFP cassette (HindIII/EcoRI) Hide
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1 AAGCTTATGG CCTCCACTCG TGTCCTCGCC TCTCGCCTGG CCTCCCAGAT GGCTGCTTCC
61 GCCAAGGTTG CCCGCCCTGC TGTCCGCGTT GCTCAGGTCA GCAAGCGCAC CATCCAGACT
121 GGCTCCCCCC TCCAGACCCT CAAGCGCACC CAGATGACCT CCATCGTCAA CGCCACCACC
181 CGCCAGGCTT TCCAGAAGCG CGCCTACTCT TCCGGATCCC CGGGTACCAG ATCTATGAGT
241 AAAGGAGAAG AACTTTTCAC TGGAGTTGTC CCAATTCTTG TTGAATTAGA TGGTGATGTT
301 AATGGGCACA AATTTTCTGT CAGTGGAGAG GGTGAAGGTG ATGCAACATA CGGAAAACTT
361 ACCCTTAAAT TTATTTGCAC TACTGGAAAA CTACCTGTTC CATGGCCAAC ACTTGTCACT
421 ACTTTCACTT ATGGTGTTCA ATGCTTTTCA AGATACCCAG ATCATATGAA ACAGCATGAC
481 TTTTTCAAGA GTGCCATGCC CGAAGGTTAT GTACAGGAAA GAACTATATT TTTCAAAGAT
541 GACGGGAACT ACAAGACACG TGCTGAAGTC AAGTTTGAAG GTGATACCCT TGTTAATAGA
601 ATCGAGTTAA AAGGTATTGA TTTTAAAGAA GATGGAAACA TTCTTGGACA CAAATTGGAA
661 TACAACTATA ACTCACACAA TGTATACATC ATGGCAGACA AACAAAAGAA TGGAATCAAA
721 GTTAACTTCA AAATTAGACA CAACATTGAA GATGGAAGCG TTCAACTAGC AGACCATTAT
781 CAACAAAATA CTCCAATTGG CGATGGCCCT GTCCTTTTAC CAGACAACCA TTACCTGTCC
841 ACACAATCTG CCCTTTCGAA AGATCCCAAC GAAAAGAGAG ACCACATGGT CCTTCTTGAG
901 TTTGTAACAG CTGCTGGGAT TACACATGGC ATGGATGAAC TATACAAATA AGAATTC
- pYES-mtGFP Hide
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1 ACGGATTAGA AGCCGCCGAG CGGGTGACAG CCCTCCGAAG GAAGACTCTC CTCCGTGCGT
61 CCTCGTCCTC ACCGGTCGCG TTCCTGAAAC GCAGATGTGC CTCGCGCCGC ACTGCTCCGA
121 ACAATAAAGA TTCTACAATA CTAGCTTTTA TGGTTATGAA GAGGAAAAAT TGGCAGTAAC
181 CTGGCCCCAC AAACCTTCAA ATGAACGAAT CAAATTAACA ACCATAGGAT GATAATGCGA
241 TTAGTTTTTT AGCCTTATTT CTGGGGTAAT TAATCAGCGA AGCGATGATT TTTGATCTAT
301 TAACAGATAT ATAAATGCAA AAACTGCATT AACCACTTTA ACTAATACTT TCAACATTTT
361 CGGTTTGTAT TACTTCTTAT TCAAATGTAA TAAAAGTATC AACAAAAAAT TGTTAATATA
421 CCTCTATACT TTAACGTCAA GGAGAAAAAA CCCCGGATCG GACTACTAGC AGCTGTAATA
481 CGACTCACTA TAGGGAATAT TAAGCTTATG GCCTCCACTC GTGTCCTCGC CTCTCGCCTG
541 GCCTCCCAGA TGGCTGCTTC CGCCAAGGTT GCCCGCCCTG CTGTCCGCGT TGCTCAGGTC
601 AGCAAGCGCA CCATCCAGAC TGGCTCCCCC CTCCAGACCC TCAAGCGCAC CCAGATGACC
661 TCCATCGTCA ACGCCACCAC CCGCCAGGCT TTCCAGAAGC GCGCCTACTC TTCCGGATCC
721 CCGGGTACCA GATCTATGAG TAAAGGAGAA GAACTTTTCA CTGGAGTTGT CCCAATTCTT
781 GTTGAATTAG ATGGTGATGT TAATGGGCAC AAATTTTCTG TCAGTGGAGA GGGTGAAGGT
841 GATGCAACAT ACGGAAAACT TACCCTTAAA TTTATTTGCA CTACTGGAAA ACTACCTGTT
901 CCATGGCCAA CACTTGTCAC TACTTTCACT TATGGTGTTC AATGCTTTTC AAGATACCCA
961 GATCATATGA AACAGCATGA CTTTTTCAAG AGTGCCATGC CCGAAGGTTA TGTACAGGAA
1021 AGAACTATAT TTTTCAAAGA TGACGGGAAC TACAAGACAC GTGCTGAAGT CAAGTTTGAA
1081 GGTGATACCC TTGTTAATAG AATCGAGTTA AAAGGTATTG ATTTTAAAGA AGATGGAAAC
1141 ATTCTTGGAC ACAAATTGGA ATACAACTAT AACTCACACA ATGTATACAT CATGGCAGAC
1201 AAACAAAAGA ATGGAATCAA AGTTAACTTC AAAATTAGAC ACAACATTGA AGATGGAAGC
1261 GTTCAACTAG CAGACCATTA TCAACAAAAT ACTCCAATTG GCGATGGCCC TGTCCTTTTA
1321 CCAGACAACC ATTACCTGTC CACACAATCT GCCCTTTCGA AAGATCCCAA CGAAAAGAGA
1381 GACCACATGG TCCTTCTTGA GTTTGTAACA GCTGCTGGGA TTACACATGG CATGGATGAA
1441 CTATACAAAT AAGAATTCTG CAGATATCCA TCACACTGGC GGCCGCTCGA GCATGCATCT
1501 AGAGGGCCGC ATCATGTAAT TAGTTATGTC ACGCTTACAT TCACGCCCTC CCCCCACATC
1561 CGCTCTAACC GAAAAGGAAG GAGTTAGACA ACCTGAAGTC TAGGTCCCTA TTTATTTTTT
1621 TATAGTTATG TTAGTATTAA GAACGTTATT TATATTTCAA ATTTTTCTTT TTTTTCTGTA
1681 CAGACGCGTG TACGCATGTA ACATTATACT GAAAACCTTG CTTGAGAAGG TTTTGGGACG
1741 CTCGAAGGCT TTAATTTGCG GCCCTGCATT AATGAATCGG CCAACGCGCG GGGAGAGGCG
1801 GTTTGCGTAT TGGGCGCTCT TCCGCTTCCT CGCTCACTGA CTCGCTGCGC TCGGTCGTTC
1861 GGCTGCGGCG AGCGGTATCA GCTCACTCAA AGGCGGTAAT ACGGTTATCC ACAGAATCAG
1921 GGGATAACGC AGGAAAGAAC ATGTGAGCAA AAGGCCAGCA AAAGCCCAGG AACCGTAAAA
1981 AGGCCGCGTT GCTGGCGTTT TTCCATAGGC TCCGCCCCCC TGACGAGCAT CACAAAAATC
2041 GACGCTCAAG TCAGAGGTGG CGAAACCCGA CAGGACTATA AAGATACCAG GCGTTTCCCC
2101 CTGGAAGCTC CCTCGTGCGC TCTCCTGTTC CGACCCTGCC GCTTACCGGA TACCTGTCCG
2161 CCTTTCTCCC TTCGGGAAGC GTGGCGCTTT CTCATAGCTC ACGCTGTAGG TATCTCAGTT
2221 CGGTGTAGGT CGTTCGCTCC AAGCTGGGCT GTGTGCACGA ACCCCCCGTT CAGCCCGACC
2281 GCTGCGCCTT ATCCGGTAAC TATCGTCTTG AGTCCAACCC GGTAAGACAC GACTTATCGC
2341 CACTGGCAGC AGCCACTGGT AACAGGATTA GCAGAGCGAG GTATGTAGGC GGTGCTACAG
2401 AGTTCTTGAA GTGGTGGCCT AACTACGGCT ACACTAGAAG GACAGTATTT GGTATCTGCG
2461 CTCTGCTGAA GCCAGTTACC TTCGGAAAAA GAGTTGGTAG CTCTTGATCC GGCAAACAAA
2521 CCACCGCTGG TAGCGGTGGT TTTTTTGTTT GCAAGCAGCA GATTACGCGC AGAAAAAAAG
2581 GATCTCAAGA AGATCCTTTG ATCTTTTCTA CGGGGTCTGA CGCTCAGTGG AACGAAAACT
2641 CACGTTAAGG GATTTTGGTC ATGAGATTAT CAAAAAGGAT CTTCACCTAG ATCCTTTTAA
2701 ATTAAAAATG AAGTTTTAAA TCAATCTAAA GTATATATGA GTAAACTTGG TCTGACAGTT
2761 ACCAATGCTT AATCAGTGAG GCACCTATCT CAGCGATCTG TCTATTTCGT TCATCCATAG
2821 TTGCCTGACT CCCCGTCGTG TAGATAACTA CGATACGGGA GCGCTTACCA TCTGGCCCCA
2881 GTGCTGCAAT GATACCGCGA GACCCACGCT CACCGGCTCC AGATTTATCA GCAATAAACC
2941 AGCCAGCCGG AAGGGCCGAG CGCAGAAGTG GTCCTGCAAC TTTATCCGCC TCCATTCAGT
3001 CTATTAATTG TTGCCGGGAA GCTAGAGTAA GTAGTTCGCC AGTTAATAGT TTGCGCAACG
3061 TTGTTGGCAT TGCTACAGGC ATCGTGGTGT CACTCTCGTC GTTTGGTATG GCTTCATTCA
3121 GCTCCGGTTC CCAACGATCA AGGCGAGTTA CATGATCCCC CATGTTGTGC AAAAAAGCGG
3181 TTAGCTCCTT CGGTCCTCCG ATCGTTGTCA GAAGTAAGTT GGCCGCAGTG TTATCACTCA
3241 TGGTTATGGC AGCACTGCAT AATTCTCTTA CTGTCATGCC ATCCGTAAGA TGCTTTTCTG
3301 TGACTGGTGA GTACTCAACC AAGTCATTCT GAGAATAGTG TATGCGGCGA CCGAGTTGCT
3361 CTTGCCCGGC GTCAATACGG GATAATAGTG TATCACATAG CAGAACTTTA AAAGTGCTCA
3421 TCATTGGAAA ACGTTCTTCG GGGCGAAAAC TCTCAAGGAT CTTACCGCTG TTGAGATCCA
3481 GTTCGATGTA ACCCACTCGT GCACCCAACT GATCTTCAGC ATCTTTTACT TTCACCAGCG
3541 TTTCTGGGTG AGCAAAAACA GGAAGGCAAA ATGCCGCAAA AAAGGGAATA AGGGCGACAC
3601 GGAAATGTTG AATACTCATA CTCTTCCTTT TTCAATGGGT AATAACTGAT ATAATTAAAT
3661 TGAAGCTCTA ATTTGTGAGT TTAGTATACA TGCATTTACT TATAATACAG TTTTTTAGTT
3721 TTGCTGGCCG CATCTTCTCA AATATGCTTC CCAGCCTGCT TTTCTGTAAC GTTCACCCTC
3781 TACCTTAGCA TCCCTTCCCT TTGCAAATAG TCCTCTTCCA ACAATAATAA TGTCAGATCC
3841 TGTAGAGACC ACATCATCCA CGGTTCTATA CTGTTGACCC AATGCGTCTC CCTTGTCATC
3901 TAAACCCACA CCGGGTGTCA TAATCAACCA ATCGTAACCT TCATCTCTTC CACCCATGTC
3961 TCTTTGAGCA ATAAAGCCGA TAACAAAATC TTTGTCGCTC TTCGCAATGT CAACAGTACC
4021 CTTAGTATAT TCTCCAGTAG ATAGGGAGCC CTTGCATGAC AATTCTGCTA ACATCAAAAG
4081 GCCTCTAGGT TCCTTTGTTA CTTCTTCTGC CGCCTGCTTC AAACCGCTAA CAATACCTGG
4141 GCCCACCACA CCGTGTGCAT TCGTAATGTC TGCCCATTCT GCTATTCTGT ATACACCCGC
4201 AGAGTACTGC AATTTGACTG TATTACCAAT GTCAGCAAAT TTTCTGTCTT CGAAGAGTAA
4261 AAAATTGTAC TTGGCGGATA ATGCCTTTAG CGGCTTAACT GTGCCCTCCA TGGAAAAATC
4321 AGTCAAGATA TCCACATGTG TTTTTAGTAA ACAAATTTTG GGACCTAATG CTTCAACTAA
4381 CTCCAGTAAT TCCTTGGTGG TACGAACATC CAATGAAGCA CACAAGTTTG TTTGCTTTTC
4441 GTGCATGATA TTAAATAGCT TGGCAGCAAC AGGACTAGGA TGAGTAGCAG CACGTTCCTT
4501 ATATGTAGCT TTCGACATGA TTTATCTTCG TTTCCTGCAG GTTTTTGTTC TGTGCAGTTG
4561 GGTTAAGAAT ACTGGGCAAT TTCATGTTTC TTCAACACTA CATATGCGTA TATATACCAA
4621 TCTAAGTCTG TGCTCCTTCC TTCGTTCTTC CTTCTGTTCG GAGATTACCG AATCAAAAAA
4681 ATTTCAAAGA AACCGAAATC AAAAAAAAGA ATAAAAAAAA AATGATGAAT TGAATTGAAA
4741 AGCTAGCTTA TCGATGATAA GCTGTCAAAG ATGAGAATTA ATTCCACGGA CTATAGACTA
4801 TACTAGATAC TCCGTCTACT GTACGATACA CTTCCGCTCA GGTCCTTGTC CTTTAACGAG
4861 GCCTTACCAC TCTTTTGTTA CTCTATTGAT CCAGCTCAGC AAAGGCAGTG TGATCTAAGA
4921 TTCTATCTTC GCGATGTAGT AAAACTAGCT AGACCGAGAA AGAGACTAGA AATGCAAAAG
4981 GCACTTCTAC AATGGCTGCC ATCATTATTA TCCGATGTGA CGCTGCAGCT TCTCAATGAT
5041 ATTCGAATAC GCTTTGAGGA GATACAGCCT AATATCCGAC AAACTGTTTT ACAGATTTAC
5101 GATCGTACTT GTTACCCATC ATTGAATTTT GAACATCCGA ACCTGGGAGT TTTCCCTGAA
5161 ACAGATAGTA TATTTGAACC TGTATAATAA TATATAGTCT AGCGCTTTAC GGAAGACAAT
5221 GTATGTATTT CGGTTCCTGG AGAAACTATT GCATCTATTG CATAGGTAAT CTTGCACGTC
5281 GCATCCCCGG TTCATTTTCT GCGTTTCCAT CTTGCACTTC AATAGCATAT CTTTGTTAAC
5341 GAAGCATCTG TGCTTCATTT TGTAGAACAA AAATGCAACG CGAGAGCGCT AATTTTTCAA
5401 ACAAAGAATC TGAGCTGCAT TTTTACAGAA CAGAAATGCA ACGCGAAAGC GCTATTTTAC
5461 CAACGAAGAA TCTGTGCTTC ATTTTTGTAA AACAAAAATG CAACGCGACG AGAGCGCTAA
5521 TTTTTCAAAC AAAGAATCTG AGCTGCATTT TTACAGAACA GAAATGCAAC GCGAGAGCGC
5581 TATTTTACCA ACAAAGAATC TATACTTCTT TTTTGTTCTA CAAAAATGCA TCCCGAGAGC
5641 GCTATTTTTC TAACAAAGCA TCTTAGATTA CTTTTTTTCT CCTTTGTGCG CTCTATAATG
5701 CAGTCTCTTG ATAACTTTTT GCACTGTAGG TCCGTTAAGG TTAGAAGAAG GCTACTTTGG
5761 TGTCTATTTT CTCTTCCATA AAAAAAGCCT GACTCCACTT CCCGCGTTTA CTGATTACTA
5821 GCGAAGCTGC GGGTGCATTT TTTCAAGATA AAGGCATCCC CGATTATATT CTATACCGAT
5881 GTGGATTGCG CATACTTTGT GAACAGAAAG TGATAGCGTT GATGATTCTT CATTGGTCAG
5941 AAAATTATGA ACGGTTTCTT CTATTTTGTC TCTATATACT ACGTATAGGA AATGTTTACA
6001 TTTTCGTATT GTTTTCGATT CACTCTATGA ATAGTTCTTA CTACAATTTT TTTGTCTAAA
6061 GAGTAATACT AGAGATAAAC ATAAAAAATG TAGAGGTCGA GTTTAGATGC AAGTTCAAGG
6121 AGCGAAAGGT GGATGGGTAG GTTATATAGG GATATAGCAC AGAGATATAT AGCAAAGAGA
6181 TACTTTTGAG CAATGTTTGT GGAAGCGGTA TTCGCAATGG GAAGCTCCAC CCCGGTTGAT
6241 AATCAGAAAA GCCCCAAAAA CAGGAAGATT GTATAAGCAA ATATTTAAAT TGTAAACGTT
6301 AATATTTTGT TAAAATTCGC GTTAAATTTT TGTTAAATCA GCTCATTTTT TAACGAATAG
6361 CCCGAAATCG GCAAAATCCC TTATAAATCA AAAGAATAGA CCGAGATAGG GTTGAGTGTT
6421 GTTCCAGTTT CCAACAAGAG TCCACTATTA AAGAACGTGG ACTCCAACGT CAAAGGGCGA
6481 AAAAGGGTCT ATCAGGGCGA TGGCCCACTA CGTGAACCAT CACCCTAATC AAGTTTTTTG
6541 GGGTCGAGGT GCCGTAAAGC AGTAAATCGG AAGGGTAAAC GGATGCCCCC ATTTAGAGCT
6601 TGACGGGGAA AGCCGGCGAA CGTGGCGAGA AAGGAAGGGA AGAAAGCGAA AGGAGCGGGG
6661 GCTAGGGCGG TGGGAAGTGT AGGGGTCACG CTGGGCGTAA CCACCACACC CGCCGCGCTT
6721 AATGGGGCGC TACAGGGCGC GTGGGGATGA TCCACTAGT