LOCUS       sequence-347423-        8304 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent reporter for HAP complex activity.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8304)
  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 8304)
  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-CCAAT-YFP
FEATURES             Location/Qualifiers
     source          1..8304
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..22
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(37..42,43..60,61..67)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    75..91
                     /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     80..102
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     99..115
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     99..115
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        136..154
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        442..849
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             2975..3691
                     /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(3693..3709)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3693..3709)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(5017..5036)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5018..5036)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5043..5060)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5043..5059)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5052..5074)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5200..5655
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5287..5306)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5497..5518
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6011..6030)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6130..6152
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6190..6208)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6293..6397
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6398..6466,6467..7258)
                     /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(6616..6635)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7429..8017
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7918..7937
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8171..8188
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
       61 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga
      121 ttacgccaag ctcggaatta accctcacta aagggaacaa aagctgggcc ggccatttaa
      181 atgtttaaac ggccacgctg gccgaggaga acttctagta tatctacata cctaatatta
      241 ttgccttatt aaaaatggaa tcccaacgat tacatcaaaa tccacattct cttcaaaatc
      301 aattgtcctg tacttccttg ttcatgtgtg ttcaaaaacg ttatatttat aggataatta
      361 tactctattt ctcaacaagt aattggttgt ttggccgagc ggtctaaggc gcctgattca
      421 agaaatatct tgaccgcagt taactgtggg aatactcagg tatcgtaaga tgcaagagtt
      481 cgaatctctt agcaaccatt atttttttcc tcaacataac gagaacacac aggggcgcta
      541 tcgcacagaa tcaaattcga tgactggaaa ttttttgtta atttcagagg tcgcctgacg
      601 catatacctt tttcaactga aaaattggga gaaaaaggaa aggtgagagc gccggaaccg
      661 gcttttcata tagaatagag aagcgttcat gactaaatgc ttgcatcaca atacttgaag
      721 ttgacaatat tatttaagga cctattgttt tttccaatag gtggttagca atcgtcttac
      781 tttctaactt ttcttacctt ttacatttca gcaatatata tatatatatt tcaaggatat
      841 accattctaa tgtctgcccc taagttacca atagatagcg gtttaatata aggtttcaac
      901 aagaaatatt ttgtgaaaat caaaagttaa tagtttcact gtcgaaaata tgaaagttta
      961 agttttcagt tcatcccatt gtagttatca aatttctgtg tttcccgtat atgtgtgctc
     1021 tgtattcatc gctatcttga aactggttat gatcgttgcg catgtgacct gtatcgctca
     1081 aaagagcccg gcaccggtac cgggcaaaag cccctagcaa gctaaaatga ttcaactttc
     1141 gtagtaattt gttcaaccgg tcacagtaaa gtcaatgact aacttttgaa tcgaggatgt
     1201 atttccaatt gaaagcaatg tatatataaa ccattgtgtt gaagcttgtc ttacagtttt
     1261 cttgtatcct cccattgttc atatagtact taaagtacca taatccttaa agaacaaact
     1321 tgaaagagaa tacactatac acaactaata aataaagaat aatcatggct gtgaccaaga
     1381 caattgtagt tctaccaggt gaccatgttg gtcaagaaat cactgaagag gccattaaag
     1441 ttttaaatgc tattcaggaa tgtcgtccag acaaggtcaa tttcaagttt gatcatcatt
     1501 tgatcggtgg tgctgcaatt gatgccactg gtgttccatt accagacgaa gctttggagg
     1561 cctctaagaa agctgatgcc gtgcttctgg gtgctgttgg tggtccaaaa tggggtactg
     1621 gcgctgtcag accagaacaa ggtcttttga aaatccgtaa ggagttgcaa ttgtatgcta
     1681 atctaagacc atgtaatttt gcatctgatt ccttactaga tctatcgcca ttgaagcctg
     1741 aaattgcaag aggtacagat ttcgtcgttg ttagagaact agtgggtggt atttatttcg
     1801 gggagagaaa agaagatgaa ggtgatggtg tcgcctggga tagcgaaaag tattctgtgc
     1861 ccgaagttca aagaatcact agaatggctg ctttcatggc tctgcaacac aacccacctt
     1921 tacctatttg gtctctagat aaagccaacg ttctagcatc atcaagatta tggagaaaaa
     1981 cagtggaaga gacgattaag aatgaattcc cacaattgac tgttcaacat caattgattg
     2041 attccgctgc tatgatcttg gttaagaacc caactcatct aaatggtatt attatcacaa
     2101 acaatatgtt tggtgatatt atctccgatg aagcctccgt gatcccaggt tcgttaggat
     2161 tacttccatc tgcctccttg gcatctcttc cagataagaa tacggcattt ggtttatatg
     2221 agccatgtca tggctctgct ccagacttac caaagggaaa ggttaacccc gtagcaacaa
     2281 tcctatctgc tgctatgatg ttgaagttgt ctctggacct gttcgaagaa ggtgaaatta
     2341 tagaacaagc agtcaagaaa gtgttggatt ctggtatcag aacagcggat ctaaagggta
     2401 ccaactctac tactgaggtc ggtgatgcag tagcgaaagc tgtcagggaa ctattagctt
     2461 agacttagta gatgtacagt tgcatacata agtctgaaca gaaagaagta aatatcatat
     2521 ctagttcggt ttagaaatca gtgactatat tggttttgac cagatcgcaa tccttgcaag
     2581 catttagtaa tgactatcaa accggtgaat catctgataa ggctatatta gcttgtgcat
     2641 tcgcatgtat cgggaaacga acggtaccgg gcccggccaa tcaggccaat caggccaatc
     2701 atgattggcc ggatccccgt cgagcagatc cgccaggcgt gtatatatag cgtggatggc
     2761 caggcaactt tagtgctgac acatacaggc atatatatat gtgtgcgacg acacatgatc
     2821 atatggcatg catgtgctct gtatgtatat aaaactcttg ttttcttctt ttctctaaat
     2881 attctttcct tatacattag gacctttgca gcataaatta ctatacttct atagacacac
     2941 aaacacaaat acacacacta aattaatact cgagatgtct aaaggtgaag aattattcac
     3001 tggtgttgtc ccaattttgg ttgaattaga tggtgatgtt aatggtcaca aattttctgt
     3061 ctccggtgaa ggtgaaggtg atgctactta cggtaaattg accttaaaat tgatttgtac
     3121 tactggtaaa ttgccagttc catggccaac cttagtcact actttaggtt atggtttgca
     3181 atgttttgct agatacccag atcatatgaa acaacatgac tttttcaagt ctgccatgcc
     3241 agaaggttat gttcaagaaa gaactatttt tttcaaagat gacggtaact acaagaccag
     3301 agctgaagtc aagtttgaag gtgatacctt agttaataga atcgaattaa aaggtattga
     3361 ttttaaagaa gatggtaaca ttttaggtca caaattggaa tacaactata actctcacaa
     3421 tgtttacatc actgctgaca aacaaaagaa tggtatcaaa gctaacttca aaattagaca
     3481 caacattgaa gatggtggtg ttcaattagc tgaccattat caacaaaata ctccaattgg
     3541 tgatggtcca gtcttgttac cagacaacca ttacttatcc tatcaatctg ccttatccaa
     3601 agatccaaac gaaaagagag accacatggt cttgttagaa tttgttactg ctgctggtat
     3661 tacccatggt atggatgaat tgtacaaata aggatccact agttctagac acctgcaaca
     3721 tgcgtgagta aataagcggc cgccaccgcg gtggagctct aagcaaatag ctaaattata
     3781 tacgaattaa tattatgatt aagtgtttac gtgagtgcga tatttttatt actatcttat
     3841 acagttgtat atactctata aaatgagttg tctattaatt aacgcgatga atgctttctg
     3901 ggtttacctc tccaacaact ctagtttact tctcaataca ttcaattgta tttgatttgt
     3961 caatacttca tcattaatca attctatagt tttgtttttc tcgtttattt ccaaatttaa
     4021 tgcatcaatt ttattattca atttgtcgtt gattttggtt aatgatttta tggtttgatc
     4081 tctggcattg attgtttgtg ttagtttttc attattgata attaaattat ttaagttagt
     4141 tatcaactcg gtgttttcaa gtttcaagtt ttcaatttct ttagagttta ttagatttgt
     4201 caaagtttct gaattgcttg attggtcctg tagaagagta tttgttgttg tggataattg
     4261 attcaatttt tgagacaatt gctggaaggc gttgaaatat ctagcatcaa tctcatggtt
     4321 tttttcccga gagtctcgta gattcaattg ttttaatata tcttgggacc actcttgatt
     4381 tgaactcatg gaaattaaac tgggtgttgt gttgtggtgt aatgattgta ccccctttgc
     4441 ttataattgt gtggccaccg aagtcggtga tgctgtcgcc gaagaagtta agaaaatcct
     4501 tgcttaaaaa gattctcttt ttttatgata tttgtacata aactttataa atgaaattca
     4561 taatagaaac gacacgaaat tacaaaatgg aatatgttca tagggtagac gaaactatat
     4621 acgcaatcta catacattta tcaagaagga gaaaaaggag gatgtaaagg aatacaggta
     4681 agcaaattga tactaatggc tcaacgtgat aaggaaaaag aattgcactt taacattaat
     4741 attgacaagg aggagggcac cacacaaaaa gttaggtgta acagaaaatc atgaaactat
     4801 gattcctaat ttatatattg gaggattttc tctaaaaaaa aaaaaataca acaaataaaa
     4861 aacactcaat gacctgacca tttgatggag tttaagtcaa tacctccttg aaccatttcc
     4921 cataatggtg aaagttccct caagaatttt actctgtcag aaacggcctt aacgacgtag
     4981 gccggccatt taaatgttta aacggccacg ctggccccct atagtgagtc gtattacaat
     5041 tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat
     5101 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat
     5161 cgcccttccc aacagttgcg cagcctgaat ggcgaatgga cgcgccctgt agcggcgcat
     5221 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     5281 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     5341 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     5401 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     5461 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     5521 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     5581 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     5641 taacgcttac aatttcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     5701 ccgcataggg taataactga tataattaaa ttgaagctct aatttgtgag tttagtatta
     5761 ggatctctac ccttggcgaa aagtcctctg ccaacaatga tgatatctga tccaccactt
     5821 acaacttcgt cgacggttct gtactgctga cccaatatgc gtatatatac caatctaagt
     5881 ctgtgctcct tccttcgttc ttccttctgt tcggagatta ccgaatcaaa aaaatttcaa
     5941 ggaaaccgaa atcaaaaaaa agaataaaaa aaaaatgatg aattgaaaag gtggtatggt
     6001 gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa
     6061 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     6121 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     6181 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
     6241 cttagacgtc ggcgcgccga ggacgtcagg tggcactttt cggggaaatg tgcgcggaac
     6301 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     6361 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     6421 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     6481 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     6541 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     6601 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     6661 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     6721 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     6781 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     6841 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     6901 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     6961 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     7021 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     7081 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     7141 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     7201 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     7261 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     7321 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     7381 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     7441 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     7501 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     7561 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     7621 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     7681 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     7741 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     7801 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     7861 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     7921 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     7981 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     8041 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     8101 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     8161 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
     8221 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
     8281 agcgggcagt gagcgcaacg caat
//
