LOCUS       sequence-321922-        8496 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gal-inducible yeast expression of the mitochondrial ""clogger"" 
            protein b2(167)delta(19)-DHFR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8496)
  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 8496)
  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: pYX233-b2delta-DHFR
FEATURES             Location/Qualifiers
     source          1..8496
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..665
                     /gene="GAL1 and GAL10 genes of S. cerevisiae"
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
     protein_bind    217..334
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     629..652
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             1147..1710
                     /codon_start=1
                     /gene="Mus musculus Dhfr"
                     /product="mouse dihydrofolate reductase"
                     /label=DHFR
                     /translation="MVRPLNCIVAVSQNMGIGKNGDLPWPPLRNEFKYFQRMTTTSSVE
                     GKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELKEPPRGAHFLAKSLDDALRLIEQP
                     ELASKVDMVWIVGGSSVYQEAMNQPGHLRLFVTRIMQEFESDTFFPEIDLGKYKLLPEY
                     PGVLSEVQEEKGIKYKFEVYEKKD"
     CDS             1821..1847
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(1975..1994)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1976..1994)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2001..2018)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2001..2017)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2010..2032)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2158..2613
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2245..2264)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2455..2476
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2716..3390)
                     /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(3391..3672)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(3693..3712)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3812..3834
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3872..3890)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     3952..3968
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3952..3968
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     rep_origin      4528..5870
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(5491..5538)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     promoter        6147..6251
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6252..6320,6321..7112)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFVAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6470..6489)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7283..7871
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7772..7791
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8025..8042
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8159..8180
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8195..8200,8201..8218,8219..8225)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8233..8249
                     /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     8238..8260
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8257..8273
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8257..8273
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        8294..8312
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 tttcaaaaat tcttactttt tttttggatg gacgcaaaga agtttaataa tcatattaca
       61 tggcattacc accatataca tatccatata catatccata tctaatctta cttatatgtt
      121 gtggaaatgt aaagagcccc attatcttag cctaaaaaaa ccttctcttt ggaactttca
      181 gtaatacgct taactgctca ttgctatatt gaagtacgga ttagaagccg ccgagcgggt
      241 gacagccctc cgaaggaaga ctctcctccg tgcgtcctcg tcttcaccgg tcgcgttcct
      301 gaaacgcaga tgtgcctcgc gccgcactgc tccgaacaat aaagattcta caatactagc
      361 ttttatggtt atgaagagga aaaattggca gtaacctggc cccacaaacc ttcaaatgaa
      421 cgaatcaaat taacaaccat aggatgataa tgcgattagt tttttagcct tatttctggg
      481 gtaattaatc agcgaagcga tgatttttga tctattaaca gatatataaa tgcaaaaact
      541 gcataaccac tttaactaat actttcaaca ttttcggttt gtattacttc ttattcaaat
      601 gtaataaaag tatcaacaaa aaattgttaa tatacctcta tactttaacg tcaaggagaa
      661 aaaaccccgg atcgaattcc caaacaaaag tagtcaatgc taaaatacaa acctttacta
      721 aaaatctcga agaactgtga ggctgctatc ctcagagcgt ctaagactag attgaacaca
      781 atccgcgcgt acggttctac cgttccaaaa tccaagtcgt tcgaacaaga ctctagttcc
      841 gtggcgtatc taaactggca taatggccaa atagacaacg agccgaaact ggatatgaat
      901 aaacaaaaga tttcgcccgc tgaagttgcc aagcataaca agcccgatga ttgttgggtt
      961 gtgatcaatg gttacgtata cgacttaacg cgattcctac caaatcatcc aggtgggcag
     1021 gatgttatca agtttaatgc cgggaaagat gtcactgcta tttttgaacc attacatgct
     1081 cctaatgtca tcgataagta tatagctccc gagaaaaaat tgggtcccct tcaaggatcc
     1141 ggcatcatgg ttcgaccatt gaactgcatc gtcgccgtgt cccaaaatat ggggattggc
     1201 aagaacggag acctaccctg gcctccgctc aggaacgagt tcaagtactt ccaaagaatg
     1261 accacaacct cttcagtgga aggtaaacag aatctggtga ttatgggtag gaaaacctgg
     1321 ttctccattc ctgagaagaa tcgaccttta aaggacagaa ttaatatagt tctcagtaga
     1381 gaactcaaag aaccaccacg aggagctcat tttcttgcca aaagtttgga tgatgcctta
     1441 agacttattg aacaaccgga attggcaagt aaagtagaca tggtttggat agtcggaggc
     1501 agttctgttt accaggaagc catgaatcaa ccaggccacc tcagactctt tgtgacaagg
     1561 atcatgcagg aatttgaaag tgacacgttt ttcccagaaa ttgatttggg gaaatataaa
     1621 cttctcccag aatacccagg cgtcctctct gaggtccagg aggaaaaagg catcaagtat
     1681 aagtttgaag tctacgagaa gaaagactaa caggaagatg ctttcaagtt ctctgctccc
     1741 ctcctaaagc tatgcatttt tataagacca tgggactttt gctggcttta gatccggcca
     1801 agcttacgcg tcgacccggg tatccgtatg atgtgcctga ctacgcatga tatctcgagc
     1861 tcagctagct aactgaataa ggaacaatga acgtttttcc tttctcttgt tcctagtatt
     1921 aatgactgac cgatacatcc cttttttttt ttgtctttgt ctagctccaa ttcgccctat
     1981 agtgagtcgt attacaattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
     2041 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
     2101 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggacg
     2161 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     2221 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     2281 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     2341 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     2401 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     2461 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     2521 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     2581 cgaattttaa caaaatatta acgcttacaa tttcctgatg cggtattttc tccttacgca
     2641 tctgtgcggt atttcacacc gcatagatcg gcaagtgcac aaacaatact taaataaata
     2701 ctactcagta ataacctatt tcttagcatt tttgacgaaa tttgctattt tgttagagtc
     2761 ttttacacca tttgtctcca cacctccgct tacatcaaca ccaataacgc catttaatct
     2821 aagcgcatca ccaacatttt ctggcgtcag tccaccagct aacataaaat gtaaactttc
     2881 ggggctctct tgccttccaa cccagtcaga aatcgagttc caatccaaaa gttcacctgt
     2941 cccacctgct tctgaatcaa acaagggaat aaacgaatga ggtttctgtg aagctgcact
     3001 gagtagtatg ttgcagtctt ttggaaatac gagtctttta ataactggca aaccgaggaa
     3061 ctcttggtat tcttgccacg actcatctcc atgcagttgg actatatcaa tgccgtaatc
     3121 attgaccaga gccaaaacat cctccttagg ttgattacga aacacgccaa ccaagtattt
     3181 cggagtgcct gaactatttt tatatgcttt tacaagactt gaaattttcc ttgcaataac
     3241 cgggtcaatt gttctctttc tattgggcac acatataata cccagcaagt cagcatcgga
     3301 atctagagca cattctgcgg cctctgtgct ctgcaagccg caaactttca ccaatggacc
     3361 agaactacct gtgaaattaa taacagacat actccaagct gcctttgtgt gcttaatcac
     3421 gtatactcac gtgctcaata gtcaccaatg ccctccctct tggccctctc cttttctttt
     3481 ttcgaccgaa ttaattctta atcggcaaaa aaagaaaagc tccggatcaa gattgtacgt
     3541 aaggtgacaa gctatttttc aataaagaat atcttccact actgccatct ggcgtcataa
     3601 ctgcaaagta cacatatatt acgatgctgt tctattaaat gcttcctata ttatatatat
     3661 agtaatgtcg ttgatctatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     3721 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     3781 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     3841 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt
     3901 aatgtcatga taataatggt ttcttagacg tactagtgag agtgcggccg ctctagaact
     3961 agtggatcaa ttccacggac tatagactat actagtatac tccgtctact gtacgataca
     4021 cttccgctca ggtccttgtc ctttaacgag gccttaccac tcttttgtta ctctattgat
     4081 ccagctcagc aaaggcagtg tgatctaaga ttctatcttc gcgatgtagt aaaactagct
     4141 agaccgagaa agagactaga aatgcaaaag gcacttctac aatggctgcc atcattatta
     4201 tccgatgtga cgctgcagct tctcaatgat attcgaatac gctttgagga gatacagcct
     4261 aatatccgac aaactgtttt acagatttac gatcgtactt gttacccatc attgaatttt
     4321 gaacatccga acctgggagt tttccctgaa acagatagta tatttgaacc tgtataataa
     4381 tatatagtct agcgctttac ggaagacaat gtatgtattt cggttcctgg agaaactatt
     4441 gcatctattg cataggtaat cttgcacgtc gcatccccgg ttcattttct gcgtttccat
     4501 cttgcacttc aatagcatat ctttgttaac gaagcatctg tgcttcattt tgtagaacaa
     4561 aaatgcaacg cgagagcgct aatttttcaa acaaagaatc tgagctgcat ttttacagaa
     4621 cagaaatgca acgcgaaagc gctattttac caacgaagaa tctgtgcttc atttttgtaa
     4681 aacaaaaatg caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc tgcattttta
     4741 cagaacagaa atgcaacgcg agagcgctat tttaccaaca aagaatctat acttcttttt
     4801 tgttctacaa aaatgcatcc cgagagcgct atttttctaa caaagcatct tagattactt
     4861 tttttctcct ttgtgcgctc tataatgcag tctcttgata actttttgca ctgtaggtcc
     4921 gttaaggtta gaagaaggct actttggtgt ctattttctc ttccataaaa aaagcctgac
     4981 tccacttccc gcgtttactg attactagcg aagctgcggg tgcatttttt caagataaag
     5041 gcatccccga ttatattcta taccgatgtg gattgcgcat actttgtgaa cagaaagtga
     5101 tagcgttgat gattcttcat tggtcagaaa attatgaacg gtttcttcta ttttgtctct
     5161 atatactacg tataggaaat gtttacattt tcgtattgtt ttcgattcac tctatgaata
     5221 gttcttacta caattttttt gtctaaagag taatactaga gataaacata aaaaatgtag
     5281 aggtcgagtt tagatgcaag ttcaaggagc gaaaggtgga tgggtaggtt atatagggat
     5341 atagcacaga gatatatagc aaagagatac ttttgagcaa tgtttgtgga agcggtattc
     5401 gcaatatttt agtagctcgt tacagtccgg tgcgtttttg gttttttgaa agtgcgtctt
     5461 cagagcgctt ttggttttca aaagcgctct gaagttccta tactttctag agaataggaa
     5521 cttcggaata ggaacttcaa agcgtttccg aaaacgagcg cttccgaaaa tgcaacgcga
     5581 gctgcgcaca tacagctcac tgttcacgtc gcacctatat ctgcgtgttg cctgtatata
     5641 tatatacatg agaagaacgg catagtgcgt gtttatgctt aaatgcgtac ttatatgcgt
     5701 ctatttatgt aggatgaaag gtagtctagt acctcctgtg atattatccc attccatgcg
     5761 gggtatcgta tgcttccttc agcactaccc tttagctgtt ctatatgctg ccactcctca
     5821 attggattag tctcatcctt caatgctatc atttcctttg atattggatc atatgcatag
     5881 taccgagaaa ctagtgcgaa gtagtgatca ggtattgctg ttatctgatg agtatacgtt
     5941 gtcctggcca cggcagaagc acgcttatcg ctccaatttc ccacaacatt agtcaactcc
     6001 gttaggccct tcattgaaag aaatgaggtc atcaaatgtc ttccaatgtg agattttggg
     6061 ccatttttta tagcaaagat tgaataaggc gcatttttct tcaaagctgc ggccgcacgt
     6121 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     6181 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     6241 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgtggcat
     6301 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     6361 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     6421 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     6481 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     6541 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     6601 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     6661 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     6721 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     6781 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     6841 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     6901 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     6961 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     7021 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     7081 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     7141 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     7201 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     7261 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     7321 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     7381 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     7441 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     7501 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     7561 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     7621 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     7681 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     7741 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     7801 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     7861 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     7921 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     7981 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     8041 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     8101 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     8161 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
     8221 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt
     8281 acgccaagct cgaaattaac cctcactaaa gggaacaaaa gctggtaccg ggccggccgt
     8341 cggaccaatt cgacaggtta tcagcaacaa cacagtcata tccattctca attagctcta
     8401 ccacagtgtg tgaaccaatg tatccagcac cacctgtaac caaaacaatt ttagaagtac
     8461 tttcactttg taactgagct gtcatttata ttgaat
//
