LOCUS       sequence-206054-        7210 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for yeast expression of GST encoded by non-preferred codons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7210)
  AUTHORS   Josse L, Sampson CDD, Tuite MF, Howland K, von der Haar T
  TITLE     Codon-Dependent Translational Accuracy Controls Protein Quality in 
            Escherichia coli but not in Saccharomyces cerevisiae
  JOURNAL   bioRxiv 200006
REFERENCE   2  (bases 1 to 7210)
  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"; journalName: "bioRxiv 
            200006"
COMMENT     Sequence Label: pTH846-gst-dissc
FEATURES             Location/Qualifiers
     source          1..7210
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        215..858
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     751..774
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     CDS             869..892
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             893..1546
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="SPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKKF
                     ELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRYG
                     VSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVLY
                     MDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     terminator      1709..1896
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1764..1788)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(1946..1962)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1946..1962)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1959..1981)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1970..1986
                     /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)."
     promoter        complement(join(1994..2000,2001..2018,2019..2024))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2039..2060
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2177..2194)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2348..2936)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2428..2447)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3107..3898,3899..3967))
                     /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     3730..3749
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3968..4072)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4140..4158
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4183..4398
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             4399..5202
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      5313..6659
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      complement(6684..6983)
                     /label=ADH2 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 2 (ADH2) gene"
ORIGIN
        1 ttagagagta gtgtgcgtga atgaaggaag gaaaaagttt cgtgtgcttc gagatacccc
       61 tcatcagctc tggaacaacg acatctgttg gtgctgtctt tgtcgttaat tttttccttt
      121 agtgtcttcc atcatttttt tgtcattgcg gatatggtga gacaacaacg ggggagagag
      181 aaaagaaaaa aaaagaaaag aatccgggag tttatcatta tcaatactgc catttcaaag
      241 aatacgtaaa taattaatag tagtgatttt cctaacttta tttagtcaaa aaattagcct
      301 tttaattctg ctgtaacccg tacatgccca aaataggggg cgggttacac agaatatata
      361 acatcgtagg tgtctgggtg aacagtttat tcctggcatc cactaaatat aatggagccc
      421 gctttttaag ctggcatcca gaaaaaaaaa gaatcccagc accaaaatat tgttttcttc
      481 accaaccatc agttcatagg tccattctct tagcgcaact acagagaaca ggggcacaaa
      541 caggcaaaaa acgggcacaa cctcaatgga gtgatgcaac ctgcctggag taaatgatga
      601 cacaaggcaa ttgacccacg catgtatcta tctcattttc ttacaccttc tattaccttc
      661 tgctctctct gatttggaaa aagctgaaaa aaaaggttga aaccagttcc ctgaaattat
      721 tcccctactt gactaataag tatataaaga cggtaggtat tgattgtaat tctgtaaatc
      781 tatttcttaa acttcttaaa ttctactttt atagttagtc ttttttttag ttttaaaaca
      841 ccaagaactt agtttcgacg gatccatgca ccatcaccat caccatcacc attcgcctat
      901 actcgggtat tggaagataa aggggctcgt acagcctaca aggctcctcc tcgagtatct
      961 cgaggagaag tatgaggagc acctctatga gagggatgag ggggataagt ggaggaataa
     1021 gaagttcgag ctcgggctcg agttccctaa tctcccttat tatatagatg gggatgtaaa
     1081 gctcacacag tcgatggcaa taataaggta tatagcagat aagcacaata tgctcggggg
     1141 gtgccctaag gagagggcag agatatcgat gctcgagggg gcagtactcg atataaggta
     1201 tggggtatcg aggatagcat attcgaagga tttcgagaca ctcaaggtag atttcctctc
     1261 gaagctccct gagatgctca agatgttcga ggataggctc tgccacaaga catatctcaa
     1321 tggggatcac gtaacacacc ctgatttcat gctctatgat gcactcgatg tagtactcta
     1381 tatggaccct atgtgcctcg atgcattccc taagctcgta tgcttcaaga agaggataga
     1441 ggcaatacct cagatagata agtatctcaa gtcgtcgaag tatatagcat ggcctctcca
     1501 ggggtggcag gcaacattcg ggggggggga tcaccctcct aagtaattct agagtcgacc
     1561 tgcaggcatg caagctttgg acttcttcgc cagaggtttg gtcaagtctc caatcaaggt
     1621 tgtcggcttg tctaccttgc cagaaattta cgaaaagatg gaaaagggtc aaatcgttgg
     1681 tagatacgtt gttgacactt ctaaataagc gaatttctta tgatttatga tttttattat
     1741 taaataagtt ataaaaaaaa taagtgtata caaattttaa agtgactctt aggttttaaa
     1801 acgaaaattc ttattcttga gtaactcttt cctgtaggtc aggttgcttt ctcaggtata
     1861 gcatgaggtc gctcttattg accacacctc taccggcatg ccggtcgaaa ttcccctacc
     1921 ctatgaaaag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     1981 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     2041 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     2101 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     2161 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     2221 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     2281 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     2341 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     2401 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     2461 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     2521 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     2581 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     2641 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     2701 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     2761 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     2821 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     2881 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     2941 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     3001 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     3061 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     3121 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     3181 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     3241 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     3301 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     3361 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     3421 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     3481 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     3541 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     3601 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     3661 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     3721 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     3781 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     3841 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     3901 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     3961 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     4021 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     4081 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     4141 ataggcgtat cacgaggccc tttcgtcttc aagaattagc ttttcaattc atcatttttt
     4201 ttttattctt ttttttgatt tcggtttcct tgaaattttt ttgattcggt aatctccgaa
     4261 cagaaggaag aacgaaggaa ggagcacaga cttagattgg tatatatacg catatgtagt
     4321 gttgaagaaa catgaaattg cccagtattc ttaacccaac tgcacagaac aaaaacatgc
     4381 aggaaacgaa gataaatcat gtcgaaagct acatataagg aacgtgctgc tactcatcct
     4441 agtcctgttg ctgccaagct atttaatatc atgcacgaaa agcaaacaaa cttgtgtgct
     4501 tcattggatg ttcgtaccac caaggaatta ctggagttag ttgaagcatt aggtcccaaa
     4561 atttgtttac taaaaacaca tgtggatatc ttgactgatt tttccatgga gggcacagtt
     4621 aagccgctaa aggcattatc cgccaagtac aattttttac tcttcgaaga cagaaaattt
     4681 gctgacattg gtaatacagt caaattgcag tactctgcgg gtgtatacag aatagcagaa
     4741 tgggcagaca ttacgaatgc acacggtgtg gtgggcccag gtattgttag cggtttgaag
     4801 caggcggcag aagaagtaac aaaggaacct agaggccttt tgatgttagc agaattgtca
     4861 tgcaagggct ccctatctac tggagaatat actaagggta ctgttgacat tgcgaagagc
     4921 gacaaagatt ttgttatcgg ctttattgct caaagagaca tgggtggaag agatgaaggt
     4981 tacgattggt tgattatgac acccggtgtg ggtttagatg acaagggaga cgcattgggt
     5041 caacagtata gaaccgtgga tgatgtggtc tctacaggat ctgacattat tattgttgga
     5101 agaggactat ttgcaaaggg aagggatgct aaggtagagg gtgaacgtta cagaaaagca
     5161 ggctgggaag catatttgag aagatgcggc cagcaaaact aaaaaactgt attataagta
     5221 aatgcatgta tactaaactc acaaattaga gcttcaattt aattatatca gttattaccc
     5281 aattctcatg tttgacagct tatcatcgga tcgatccaat atcaaaggaa atgatagcat
     5341 tgaaggatga gactaatcca attgaggagt ggcagcatat agaacagcta aagggtagtg
     5401 ctgaaggaag catacgatac cccgcatgga atgggataat atcacaggag gtactagact
     5461 acctttcatc ctacataaat agacgcatat aagtacgcat ttaagcataa acacgcacta
     5521 tgccgttctt ctcatgtata tatatataca ggcaacacgc agatataggt gcgacgtgaa
     5581 cagtgagctg tatgtgcgca gctcgcgttg cattttcgga agcgctcgtt ttcggaaacg
     5641 ctttgaagtt cctattccga agttcctatt ctctagctag aaagtatagg aacttcagag
     5701 cgcttttgaa aaccaaaagc gctctgaaga cgcactttca aaaaaccaaa aacgcaccgg
     5761 actgtaacga gctactaaaa tattgcgaat accgcttcca caaacattgc tcaaaagtat
     5821 ctctttgcta tatatctctg tgctatatcc ctatataacc tacccatcca cctttcgctc
     5881 cttgaacttg catctaaact cgacctctac attttttatg tttatctcta gtattactct
     5941 ttagacaaaa aaattgtagt aagaactatt catagagtga atcgaaaaca atacgaaaat
     6001 gtaaacattt cctatacgta gtatatagag acaaaataga agaaaccgtt cataattttc
     6061 tgaccaatga agaatcatca acgctatcac tttctgttca caaagtatgc gcaatccaca
     6121 tcggtataga atataatcgg ggatgccttt atcttgaaaa aatgcacccg cagcttcgct
     6181 agtaatcagt aaacgcggga agtggagtca ggcttttttt atggaagaga aaatagacac
     6241 caaagtagcc ttcttctaac cttaacggac ctacagtgca aaaagttatc aagagactgc
     6301 attatagagc gcacaaagga gaaaaaaagt aatctaagat gctttgttag aaaaatagcg
     6361 ctctcgggat gcatttttgt agaacaaaaa agaagtatag attctttgtt ggtaaaatag
     6421 cgctctcgcg ttgcatttct gttctgtaaa aatgcagctc agattctttg tttgaaaaat
     6481 tagcgctctc gcgttgcatt tttgttttac aaaaatgaag cacagattct tcgttggtaa
     6541 aatagcgctt tcgcgttgca tttctgttct gtaaaaatgc agctcagatt ctttgtttga
     6601 aaaattagcg ctctcgcgtt gcatttttgt tctacaaaat gaagcacaga tgcttcgttc
     6661 cgctcgaggg catgcgaagg aaaatgagaa atatcgaggg agacgattca gaggagcagg
     6721 acaaactata accgactgtt tgttggagga tgccgtacat aacgaacact gctgaagcta
     6781 ccatgtctac agtttagagg aatgggtaca actcacaggc gagggatggt gttcactcgt
     6841 gctagcaaac gcggtgggag caaaaagtag aatattatct tttattcgtg aaacttcgaa
     6901 cactgtcatc taaagatgct atatactaat ataggcatac ttgataatga aaactataaa
     6961 tcgtaaagac ataagagatc cgcttattta gagtgtcgaa ttcgagctcg gtacccgggg
     7021 agttgattgt agcttggtat agcttgaaat attgtgcaga aaaagaaaca aggaagaaag
     7081 ggaacgagaa caatgacgag gaaacaaaag attaataatt gcaggtctat ttatacttga
     7141 tagcaagaca gcaaactttt ttttatttca aattcaagta actggaagga aggccgtata
     7201 ccgttgctca
//
