LOCUS       sequence-180676-        7276 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses mCherry-GFP1-10 in yeast cell mitochondria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 7276)
  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 7276)
  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-MTS-mCherry-GFP1-10
FEATURES             Location/Qualifiers
     source          1..7276
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        complement(4..347)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     promoter        360..1003
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     897..920
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     regulatory      1249..1258
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     terminator      2635..2882
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(2651..2669)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(2900..2919)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2901..2919)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2929..2946)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2929..2945)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2938..2960)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3086..3541
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3173..3192)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3383..3404
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3644..4318)
                     /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(4319..4600)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(4621..4640)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4740..4762
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4800..4818)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4886..4990
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4991..5059,5060..5851)
                     /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(5209..5228)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6022..6610
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6511..6530
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6764..6781
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6898..6919
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6934..6939,6940..6957,6958..6964)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6972..6988
                     /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     6977..6999
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6996..7012
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6996..7012
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7031..7051
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7033..7051
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     terminator      complement(7070..7267)
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
ORIGIN
        1 catggttgtt tatgttcgga tgtgatgtga gaactgtatc ctagcaagat tttaaaagga
       61 agtatatgaa agaagaacct cagtggcaaa tcctaacctt ttatatttct ctacaggggc
      121 gcggcgtggg gacaattcaa cgcgtctgtg aggggagcgt ttccctgctc gcaggtctgc
      181 agcgaggagc cgtaattttt gcttcgcgcc gtgcggccat caaaatgtat ggatgcaaat
      241 gattatacat ggggatgtat gggctaaatg tacgggcgac agtcacatca tgcccctgag
      301 ctgcgcacgt caagactgtc aaggagggta ttctgggcct ccatgtcgag ctcagtttat
      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 aattattccc ctacttgact aataagtata taaagacggt
      901 aggtattgat tgtaattctg taaatctatt tcttaaactt cttaaattct acttttatag
      961 ttagtctttt ttttagtttt aaaacaccag aacttagttt cgacggattc tagaactagt
     1021 atggcctcca ctcgtgtcct cgcctctcgc ctggcctccc ggatggctgc ttccgccaag
     1081 gttgcccgcc ctgctgtccg cgttgctcag gtcagcaagc gcaccatcca gactggctcc
     1141 cccctccaga ccctcaagcg cacccagatg acctccatcg tcaacgccac cacccgccag
     1201 gctttccaga agcgcgccta ctcttccgga tccggtgacg gtccggtcgc caccatggtt
     1261 tctaaaggtg aagaagataa tatggctatt attaaagaat ttatgagatt taaagttcat
     1321 atggaaggtt ctgttaatgg tcatgaattt gaaattgaag gtgaaggtga aggtagacca
     1381 tatgaaggta ctcaaactgc taaattgaaa gttactaaag gtggtccatt gccatttgct
     1441 tgggatattt tgtctccaca atttatgtat ggttctaaag cttatgttaa acatccagct
     1501 gatattccag attatttgaa attgtctttt ccagaaggtt ttaaatggga aagagttatg
     1561 aattttgaag atggtggtgt tgttactgtt actcaagatt cttctttgca agatggtgaa
     1621 tttatttata aagttaaatt gagaggtact aattttccat ctgatggtcc agttatgcaa
     1681 aaaaaaacta tgggttggga agcttcttct gaaagaatgt atccagaaga tggtgctttg
     1741 aaaggtgaaa ttaaacaaag attgaaattg aaagatggtg gtcattatga tgctgaagtt
     1801 aaaactactt ataaagctaa aaaaccagtt caattgccag gtgcttataa tgttaatatt
     1861 aaattggata ttacttctca taatgaagat tatactattg ttgaacaata tgaaagagct
     1921 gaaggtagac attctactgg tggtatggat gaattgtata aagaattcgg tggtggttct
     1981 ggcggatcta aaggtgaaga attgtttact ggtgttgttc caattttggt tgaattggat
     2041 ggtgatgtta atggtcataa attttctgtt agaggtgaag gtgaaggtga tgctactatt
     2101 ggtaaattga ctttgaaatt tatttgtact actggtaaat tgccagttcc atggccaact
     2161 ttggttacta ctttgactta tggtgttcaa tgtttttcta gatatccaga tcatatgaaa
     2221 caacatgatt tttttaaatc tgctatgcca gaaggttatg ttcaagaaag aactatttct
     2281 tttaaagatg atggtaaata taaaactaga gctgttgtta aatttgaagg tgatactttg
     2341 gttaatagaa ttgaattgaa aggtactgat tttaaagaag atggtaatat tttgggtcat
     2401 aaattggaat ataattttaa ttctcataat gtttatatta ctgctgataa acaaaaaaat
     2461 ggtattaaag ctaattttac tgttagacat aatgttgaag atggttctgt tcaattggct
     2521 gatcattatc aacaaaatac tccaattggt gatggtccag ttttgttgcc agataatcat
     2581 tatttgtcta ctcaaactgt tttgtctaaa gatccaaatg aaaaataact cgagtcatgt
     2641 aattagttat gtcacgctta cattcacgcc ctccccccac atccgctcta accgaaaagg
     2701 aaggagttag acaacctgaa gtctaggtcc ctatttattt ttttatagtt atgttagtat
     2761 taagaacgtt atttatattt caaatttttc ttttttttct gtacagacgc gtgtacgcat
     2821 gtaacattat actgaaaacc ttgcttgaga aggttttggg acgctcgaag gctttaattt
     2881 gcggccggta cccaattcgc cctatagtga gtcgtattac gcgcgctcac tggccgtcgt
     2941 tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca
     3001 tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
     3061 gttgcgcagc ctgaatggcg aatggacgcg ccctgtagcg gcgcattaag cgcggcgggt
     3121 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     3181 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     3241 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     3301 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     3361 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     3421 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     3481 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt
     3541 tcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atagatcggc
     3601 aagtgcacaa acaatactta aataaatact actcagtaat aacctatttc ttagcatttt
     3661 tgacgaaatt tgctattttg ttagagtctt ttacaccatt tgtctccaca cctccgctta
     3721 catcaacacc aataacgcca tttaatctaa gcgcatcacc aacattttct ggcgtcagtc
     3781 caccagctaa cataaaatgt aagctttcgg ggctctcttg ccttccaacc cagtcagaaa
     3841 tcgagttcca atccaaaagt tcacctgtcc cacctgcttc tgaatcaaac aagggaataa
     3901 acgaatgagg tttctgtgaa gctgcactga gtagtatgtt gcagtctttt ggaaatacga
     3961 gtcttttaat aactggcaaa ccgaggaact cttggtattc ttgccacgac tcatctccat
     4021 gcagttggac gatatcaatg ccgtaatcat tgaccagagc caaaacatcc tccttaggtt
     4081 gattacgaaa cacgccaacc aagtatttcg gagtgcctga actattttta tatgctttta
     4141 caagacttga aattttcctt gcaataaccg ggtcaattgt tctctttcta ttgggcacac
     4201 atataatacc cagcaagtca gcatcggaat ctagagcaca ttctgcggcc tctgtgctct
     4261 gcaagccgca aactttcacc aatggaccag aactacctgt gaaattaata acagacatac
     4321 tccaagctgc ctttgtgtgc ttaatcacgt atactcacgt gctcaatagt caccaatgcc
     4381 ctccctcttg gccctctcct tttctttttt cgaccgaatt aattcttaat cggcaaaaaa
     4441 agaaaagctc cggatcaaga ttgtacgtaa ggtgacaagc tatttttcaa taaagaatat
     4501 cttccactac tgccatctgg cgtcataact gcaaagtaca catatattac gatgctgttc
     4561 tattaaatgc ttcctatatt atatatatag taatgtcgtt gatctatggt gcactctcag
     4621 tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga
     4681 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     4741 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg
     4801 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     4861 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     4921 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     4981 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     5041 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     5101 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     5161 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     5221 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     5281 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     5341 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     5401 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     5461 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     5521 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     5581 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     5641 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     5701 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     5761 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     5821 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     5881 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     5941 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     6001 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     6061 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     6121 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     6181 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     6241 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     6301 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     6361 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     6421 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     6481 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     6541 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     6601 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     6661 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     6721 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     6781 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     6841 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
     6901 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
     6961 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta
     7021 cgccaagcgc gcaattaacc ctcactaaag ggaacaaaag ctggagctcc agtatagcga
     7081 ccagcattca catacgattg acgcatgata ttactttctg cgcacttaac ttcgcatctg
     7141 ggcagatgat gtcgaggcga aaaaaaatat aaatcacgct aacatttgat taaaatagaa
     7201 caactacaat ataaaaaaac tatacaaatg acaagttctt gaaaacaaga atctttttat
     7261 tgtcagtact gattag
//
