LOCUS       sequence-346797-        8317 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent reporter for Rpn4 activity.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8317)
  AUTHORS   Boos F, Kramer L, Groh C, Jung F, Haberkant P, Stein F, Wollweber F,
            Gackstatter A, Zoller E, van der Laan M, Savitski MM, Benes V, 
            Herrmann JM
  TITLE     Mitochondrial protein-induced stress triggers a global adaptive 
            transcriptional programme.
  JOURNAL   Nat Cell Biol. 2019 Apr;21(4):442-451. doi: 
            10.1038/s41556-019-0294-5. Epub 2019 Mar 18.
  PUBMED    30886345
REFERENCE   2  (bases 1 to 8317)
  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/s41556-019-0294-5"; journalName: "Nat Cell Biol"; date: 
            "2019-04"; volume: "21"; issue: "4"; pages: "442-451"
COMMENT     Sequence Label: pNH605-PACE-YFP
FEATURES             Location/Qualifiers
     source          1..8317
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        276..683
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             2815..3531
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     LICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(3533..3549)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3533..3549)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(4864..4883)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4865..4883)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4890..4907)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4890..4906)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4899..4921)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5047..5502
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5134..5153)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5344..5365
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5858..5877)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5977..5999
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6037..6055)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6140..6244
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6245..6313,6314..7105)
                     /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(6463..6482)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7276..7864
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7765..7784
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8018..8035
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8152..8173
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8188..8193,8194..8211,8212..8218)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8226..8242
                     /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     8231..8253
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8250..8266
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8250..8266
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        8287..8305
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 ggccggccat ttaaatgttt aaacggccac gctggccgag gagaacttct agtatatcta
       61 catacctaat attattgcct tattaaaaat ggaatcccaa caattacatc aaaatccaca
      121 ttctcttcaa aatcaattgt cctgtacttc cttgttcatg tgtgttcaaa aacgttatat
      181 ttataggata attatactct atttctcaac aagtaattgg ttgtttggcc gagcggtcta
      241 aggcgcctga ttcaagaaat atcttgaccg cagttaactg tgggaatact caggtatcgt
      301 aagatgcaag agttcgaatc tcttagcaac cattattttt ttcctcaaca taacgagaac
      361 acacaggggc gctatcgcac agaatcaaat tcgatgactg gaaatttttt gttaatttca
      421 gaggtcgcct gacgcatata cctttttcaa ctgaaaaatt gggagaaaaa ggaaaggtga
      481 gagcgccgga accggctttt catatagaat agagaagcgt tcatgactaa atgcttgcat
      541 cacaatactt gaagttgaca atattattta aggacctatt gttttttcca ataggtggtt
      601 agcaatcgtc ttactttcta acttttctta ccttttacat ttcagcaata tatatatata
      661 tatttcaagg atataccatt ctaatgtctg cccctaagtt accaatagat agcggtttaa
      721 tataaggttt caacaagaaa tattttgtga aaatcaaaag ttaatagttt cactgtcgaa
      781 aatatgaaag tttaagtttt cagttcatcc cattgtagtt atcaaatttc tgtgtttccc
      841 gtatatgtgt gctctgtatt catcgctatc ttgaaactgg ttatgatcgt tgcgcatgtg
      901 acctgtatcg ctcaaaagag cccggcaccg gtaccgggca aaagccccta gcaagctaaa
      961 atgattcaac tttcgtagta atttgttcaa ccggtcacag taaagtcaat gactaacttt
     1021 tgaatcgagg atgtatttcc aattgaaagc aatgtatata taaaccattg tgttgaagct
     1081 tgtcttacag ttttcttgta tcctcccatt gttcatatag tacttaaagt accataatcc
     1141 ttaaagaaca aacttgaaag agaatacact atacacaact aataaataaa gaataatcat
     1201 ggctgtgacc aagacaattg tagttctacc aggtgaccat gttggtcaag aaatcactga
     1261 agaggccatt aaagttttaa atgctattca ggaatgtcgt ccagacaagg tcaatttcaa
     1321 gtttgatcat catttgatcg gtggtgctgc aattgatgcc actggtgttc cattaccaga
     1381 cgaagctttg gaggcctcta agaaagctga tgccgtgctt ctgggtgctg ttggtggtcc
     1441 aaaatggggt actggcgctg tcagaccaga acaaggtctt ttgaaaatcc gtaaggagtt
     1501 gcaattgtat gctaatctaa gaccatgtaa ttttgcatct gattccttac tagatctatc
     1561 gccattgaag cctgaaattg caagaggtac agatttcgtc gttgttagag aactagtggg
     1621 tggtatttat ttcggggaga gaaaagaaga tgaaggtgat ggtgtcgcct gggatagcga
     1681 aaagtattct gtgcccgaag ttcaaagaat cactagaatg gctgctttca tggctctgca
     1741 acacaaccca cctttaccta tttggtctct agataaagcc aacgttctag catcatcaag
     1801 attatggaga aaaacagtgg aagagacgat taagaatgaa ttcccacaat tgactgttca
     1861 acatcaattg attgattccg ctgctatgat cttggttaag aacccaactc atctaaatgg
     1921 tattattatc acaaacaata tgtttggtga tattatctcc gatgaagcct ccgtgatccc
     1981 aggttcgtta ggattacttc catctgcctc cttggcatct cttccagata agaatacggc
     2041 atttggttta tatgagccat gtcatggctc tgctccagac ttaccaaagg gaaaggttaa
     2101 ccccgtagca acaatcctat ctgctgctat gatgttgaag ttgtctctgg acctgttcga
     2161 agaaggtgaa attatagaac aagcagtcaa gaaagtgttg gattctggta tcagaacagc
     2221 ggatctaaag ggtaccaact ctactactga ggtcggtgat gcagtagcga aagctgtcag
     2281 ggaactatta gcttagactt agtagatgta cagttgcata cataagtctg aacagaaaga
     2341 agtaaatatc atatctagtt cggtttagaa atcagtgact atattggttt tgaccagatc
     2401 gcaatccttg caagcattta gtaatgacta tcaaaccggt gaatcatctg ataaggctat
     2461 attagcttgt gcattcgcat gtatcgggaa acgaacggta ccgggcccgg tggcaaaggt
     2521 ggcaaaggtg gcaaatttgc caccggatcc ccgtcgagca gatccgccag gcgtgtatat
     2581 atagcgtgga tggccaggca actttagtgc tgacacatac aggcatatat atatgtgtgc
     2641 gacgacacat gatcatatgg catgcatgtg ctctgtatgt atataaaact cttgttttct
     2701 tcttttctct aaatattctt tccttataca ttaggacctt tgcagcataa attactatac
     2761 ttctatagac acacaaacac aaatacacac actaaattaa tactcgagaa taaaatgtct
     2821 aaaggtgaag aattattcac tggtgttgtc ccaattttgg ttgaattaga tggtgatgtt
     2881 aatggtcaca aattttctgt ctccggtgaa ggtgaaggtg atgctactta cggtaaattg
     2941 accttaaaat tgatttgtac tactggtaaa ttgccagttc catggccaac cttagtcact
     3001 actttaggtt atggtttgca atgttttgct agatacccag atcatatgaa acaacatgac
     3061 tttttcaagt ctgccatgcc agaaggttat gttcaagaaa gaactatttt tttcaaagat
     3121 gacggtaact acaagaccag agctgaagtc aagtttgaag gtgatacctt agttaataga
     3181 atcgaattaa aaggtattga ttttaaagaa gatggtaaca ttttaggtca caaattggaa
     3241 tacaactata actctcacaa tgtttacatc actgctgaca aacaaaagaa tggtatcaaa
     3301 gctaacttca aaattagaca caacattgaa gatggtggtg ttcaattagc tgaccattat
     3361 caacaaaata ctccaattgg tgatggtcca gtcttgttac cagacaacca ttacttatcc
     3421 tatcaatctg ccttatccaa agatccaaac gaaaagagag accacatggt cttgttagaa
     3481 tttgttactg ctgctggtat tacccatggt atggatgaat tgtacaaata aggatccact
     3541 agttctagac acctgcaaca tgcgtgagta aataagcggc cgccaccgcg gtggagctct
     3601 aagcaaatag ctaaattata tacgaattaa tattatgatt aagtgtttac gtgagtgcga
     3661 tatttttatt actatcttat acagttgtat atactctata aaatgagttg tctattaatt
     3721 aacgcgatga atgctttctg ggtttacctc tccaacaact ctagtttact tctcaataca
     3781 ttcaattgta tttgatttgt caatacttca tcattaatca attctatagt tttgtttttc
     3841 tcgtttattt ccaaatttaa tgcatcaatt ttattattca atttgtcgtt gattttggtt
     3901 aatgatttta tggtttgatc tctggcattg attgtttgtg ttagtttttc attattgata
     3961 attaaattat ttaagttagt tatcaactcg gtgttttcaa gtttcaagtt ttcaatttct
     4021 ttagagttta ttagatttgt caaagtttct gaattgcttg attggtcctg tagaagagta
     4081 tttgttgttg tggataattg attcaatttt tgagacaatt gctggaaggc gttgaaatat
     4141 ctagcatcaa tctcatggtt tttttcccga gagtctcgta gattcaattg ttttaatata
     4201 tcttgggacc actcttgatt tgaactcatg gaaattaaac tgggtgttgt gttgtggtgt
     4261 aatgattgta ccccctttgc ttataattgt gtggccaccg aagtcggtga tgctgtcgcc
     4321 gaagaagtta agaaaatcct tgcttaaaaa gattctcttt ttttatgata tttgtacata
     4381 aactttataa atgaaattca taatagaaac gacacgaaat tacaaaatgg aatatgttca
     4441 tagggtagac gaaactatat acgcaatcta catacattta tcaagaagga gaaaaaggag
     4501 gatgtaaagg aatacaggta agcaaattga tactaatggc tcaacgtgat aaggaaaaag
     4561 aattgcactt taacattaat attgacaagg aggagggcac cacacaaaaa gttaggtgta
     4621 acagaaaatc atgaaactat gattcctaat ttatatattg gaggattttc tctaaaaaaa
     4681 aaaaaataca acaaataaaa aacactcaat gacctgacca tttgatggag tttaagtcaa
     4741 taccttcttg aaccatttcc cataatggtg aaagttccct caagaatttt actctgtcag
     4801 aaacggcctt aacgacgtag gccggccatt taaatgttta aacggccacg ctggcccaat
     4861 tcgccctata gtgagtcgta ttacaattca ctggccgtcg ttttacaacg tcgtgactgg
     4921 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     4981 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     5041 gaatggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     5101 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     5161 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     5221 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     5281 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     5341 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     5401 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     5461 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttcctgatgc ggtattttct
     5521 ccttacgcat ctgtgcggta tttcacaccg catagggtaa taactgatat aattaaattg
     5581 aagctctaat ttgtgagttt agtattagga tctctaccct tggcgaaaag tcctctgcca
     5641 acaatgatga tatctgatcc accacttaca acttcgtcga cggttctgta ctgctgaccc
     5701 aatatgcgta tatataccaa tctaagtctg tgctccttcc ttcgttcttc cttctgttcg
     5761 gagattaccg aatcaaaaaa atttcaagga aaccgaaatc aaaaaaaaga ataaaaaaaa
     5821 aatgatgaat tgaaaaggtg gtatggtgca ctctcagtac aatctgctct gatgccgcat
     5881 agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     5941 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     6001 tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat
     6061 aggttaatgt catgataata atggtttctt agacgtcggc gcgccgagga cgtcaggtgg
     6121 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     6181 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     6241 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     6301 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     6361 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     6421 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     6481 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     6541 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     6601 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     6661 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     6721 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     6781 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     6841 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     6901 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     6961 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     7021 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     7081 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     7141 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     7201 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     7261 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     7321 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     7381 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     7441 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     7501 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     7561 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     7621 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     7681 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     7741 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     7801 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     7861 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     7921 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     7981 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     8041 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
     8101 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag
     8161 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc ctatgttgtg
     8221 tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa
     8281 gctcggaatt aaccctcact aaagggaaca aaagctg
//
