LOCUS       sequence-180669-        6823 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses GFP1-10 in the mitochondrial matrix.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 6823)
  AUTHORS   Ruan L, Zhou C, Jin E, Kucharavy A, Zhang Y, Wen Z, Florens L, Li R
  TITLE     Cytosolic proteostasis through importing of misfolded proteins into 
            mitochondria.
  JOURNAL   Nature. 2017 Mar 16;543(7645):443-446. doi: 10.1038/nature21695. 
            Epub 2017 Mar 1.
  PUBMED    28241148
REFERENCE   2  (bases 1 to 6823)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/nature21695"; journalName: "Nature"; date: "2017-03-16- 
            16"; volume: "543"; issue: "7645"; pages: "443-446"
COMMENT     Sequence Label: p404-Grx5-GFP1-10
FEATURES             Location/Qualifiers
     source          1..6823
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        complement(1..344)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     promoter        357..1000
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     894..917
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     CDS             1474..1500
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     terminator      2182..2429
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(2198..2216)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(2447..2466)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2448..2466)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2476..2493)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2476..2492)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2485..2507)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2633..3088
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2720..2739)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2930..2951
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3191..3865)
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     promoter        complement(3866..4147)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(4168..4187)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4287..4309
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4347..4365)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4433..4537
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4538..4606,4607..5398)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4756..4775)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5569..6157
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6058..6077
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6311..6328
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6445..6466
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6481..6486,6487..6504,6505..6511)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6519..6535
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     6524..6546
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6543..6559
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6543..6559
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6578..6598
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6580..6598
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     terminator      complement(6617..6814)
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
ORIGIN
        1 ggttgtttat gttcggatgt gatgtgagaa ctgtatccta gcaagatttt aaaaggaagt
       61 atatgaaaga agaacctcag tggcaaatcc taacctttta tatttctcta caggggcgcg
      121 gcgtggggac aattcaacgc gtctgtgagg ggagcgtttc cctgctcgca ggtctgcagc
      181 gaggagccgt aatttttgct tcgcgccgtg cggccatcaa aatgtatgga tgcaaatgat
      241 tatacatggg gatgtatggg ctaaatgtac gggcgacagt cacatcatgc ccctgagctg
      301 cgcacgtcaa gactgtcaag gagggtattc tgggcctcca tgtcgagctc agtttatcat
      361 tatcaatact cgccatttca aagaatacgt aaataattaa tagtagtgat tttcctaact
      421 ttatttagtc aaaaaattag ccttttaatt ctgctgtaac ccgtacatgc ccaaaatagg
      481 gggcgggtta cacagaatat ataacatcgt aggtgtctgg gtgaacagtt tattcctggc
      541 atccactaaa tataatggag cccgcttttt aagctggcat ccagaaaaaa aaagaatccc
      601 agcaccaaaa tattgttttc ttcaccaacc atcagttcat aggtccattc tcttagcgca
      661 actacagaga acaggggcac aaacaggcaa aaaacgggca caacctcaat ggagtgatgc
      721 aacctgcctg gagtaaatga tgacacaagg caattgaccc acgcatgtat ctatctcatt
      781 ttcttacacc ttctattacc ttctgctctc tctgatttgg aaaaagctga aaaaaaaggt
      841 tgaaaccagt tccctgaaat tattccccta cttgactaat aagtatataa agacggtagg
      901 tattgattgt aattctgtaa atctatttct taaacttctt aaattctact tttatagtta
      961 gtcttttttt tagttttaaa acaccagaac ttagtttcga cggattctag aactagtatg
     1021 tttctcccaa aattcaatcc cataaggtca ttttccccca tcctccgggc taagactctt
     1081 cttcgttacc aaaatcggat gtatttgagc acagagataa gaaaagctat tgaagatgcc
     1141 atcgaatcgg ctccagtggt tcttttcatg aaaggtactc ctgaatttcc caagtgtgga
     1201 ttttcaagag caaccattgg attattagga aatcaaggcg ttgagccggc caaatttgcg
     1261 gcttataatg ttttagaaga cccagagcta cgtgaaggta tcaaagagtt ttcagaatgg
     1321 ccaactattc cacagttata tgtaaacaaa gaattcattg gtggatgtga tgttattaca
     1381 agtatggcac gctctggtga attggccgat ttgctagaag aggcacaggc attggtacct
     1441 gaagaagaag aagaaaccaa agatcgtgga tcctacccat acgatgttcc agattacgct
     1501 gaattcggtg gtggttctgg cggaggttca atgtctaaag gtgaagaatt gtttactggt
     1561 gttgttccaa ttttggttga attggatggt gatgttaatg gtcataaatt ttctgttaga
     1621 ggtgaaggtg aaggtgatgc tactattggt aaattgactt tgaaatttat ttgtactact
     1681 ggtaaattgc cagttccatg gccaactttg gttactactt tgacttatgg tgttcaatgt
     1741 ttttctagat atccagatca tatgaaacaa catgattttt ttaaatctgc tatgccagaa
     1801 ggttatgttc aagaaagaac tatttctttt aaagatgatg gtaaatataa aactagagct
     1861 gttgttaaat ttgaaggtga tactttggtt aatagaattg aattgaaagg tactgatttt
     1921 aaagaagatg gtaatatttt gggtcataaa ttggaatata attttaattc tcataatgtt
     1981 tatattactg ctgataaaca aaaaaatggt attaaagcta attttactgt tagacataat
     2041 gttgaagatg gttctgttca attggctgat cattatcaac aaaatactcc aattggtgat
     2101 ggtccagttt tgttgccaga taatcattat ttgtctactc aaactgtttt gtctaaagat
     2161 ccaaatgaaa aataactcga gtcatgtaat tagttatgtc acgcttacat tcacgccctc
     2221 cccccacatc cgctctaacc gaaaaggaag gagttagaca acctgaagtc taggtcccta
     2281 tttatttttt tatagttatg ttagtattaa gaacgttatt tatatttcaa atttttcttt
     2341 tttttctgta cagacgcgtg tacgcatgta acattatact gaaaaccttg cttgagaagg
     2401 ttttgggacg ctcgaaggct ttaatttgcg gccggtaccc aattcgccct atagtgagtc
     2461 gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
     2521 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga
     2581 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggacgcgccc
     2641 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     2701 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     2761 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     2821 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     2881 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     2941 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     3001 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     3061 tttaacaaaa tattaacgct tacaatttcc tgatgcggta ttttctcctt acgcatctgt
     3121 gcggtatttc acaccgcata gatcggcaag tgcacaaaca atacttaaat aaatactact
     3181 cagtaataac ctatttctta gcatttttga cgaaatttgc tattttgtta gagtctttta
     3241 caccatttgt ctccacacct ccgcttacat caacaccaat aacgccattt aatctaagcg
     3301 catcaccaac attttctggc gtcagtccac cagctaacat aaaatgtaag ctttcggggc
     3361 tctcttgcct tccaacccag tcagaaatcg agttccaatc caaaagttca cctgtcccac
     3421 ctgcttctga atcaaacaag ggaataaacg aatgaggttt ctgtgaagct gcactgagta
     3481 gtatgttgca gtcttttgga aatacgagtc ttttaataac tggcaaaccg aggaactctt
     3541 ggtattcttg ccacgactca tctccatgca gttggacgat atcaatgccg taatcattga
     3601 ccagagccaa aacatcctcc ttaggttgat tacgaaacac gccaaccaag tatttcggag
     3661 tgcctgaact atttttatat gcttttacaa gacttgaaat tttccttgca ataaccgggt
     3721 caattgttct ctttctattg ggcacacata taatacccag caagtcagca tcggaatcta
     3781 gagcacattc tgcggcctct gtgctctgca agccgcaaac tttcaccaat ggaccagaac
     3841 tacctgtgaa attaataaca gacatactcc aagctgcctt tgtgtgctta atcacgtata
     3901 ctcacgtgct caatagtcac caatgccctc cctcttggcc ctctcctttt cttttttcga
     3961 ccgaattaat tcttaatcgg caaaaaaaga aaagctccgg atcaagattg tacgtaaggt
     4021 gacaagctat ttttcaataa agaatatctt ccactactgc catctggcgt cataactgca
     4081 aagtacacat atattacgat gctgttctat taaatgcttc ctatattata tatatagtaa
     4141 tgtcgttgat ctatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     4201 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     4261 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     4321 atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     4381 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     4441 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     4501 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     4561 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     4621 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     4681 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     4741 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     4801 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     4861 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     4921 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     4981 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     5041 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     5101 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     5161 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     5221 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     5281 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     5341 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     5401 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     5461 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     5521 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     5581 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     5641 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     5701 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     5761 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     5821 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     5881 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     5941 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     6001 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     6061 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     6121 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     6181 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     6241 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     6301 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
     6361 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
     6421 agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc
     6481 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
     6541 cacaggaaac agctatgacc atgattacgc caagcgcgca attaaccctc actaaaggga
     6601 acaaaagctg gagctccagt atagcgacca gcattcacat acgattgacg catgatatta
     6661 ctttctgcgc acttaacttc gcatctgggc agatgatgtc gaggcgaaaa aaaatataaa
     6721 tcacgctaac atttgattaa aatagaacaa ctacaatata aaaaaactat acaaatgaca
     6781 agttcttgaa aacaagaatc tttttattgt cagtactgat tag
//
