LOCUS       sequence-206073-        7210 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for yeast expression of GST encoded by 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: pTH847-gst-optsc
FEATURES             Location/Qualifiers
     source          1..7210
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        271..914
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     807..830
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     CDS             925..948
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             949..1602
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="SPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKKF
                     ELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRYG
                     VSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVLY
                     MDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     terminator      1765..1952
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1820..1844)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(2002..2018)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2002..2018)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2015..2037)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2026..2042
                     /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(2050..2056,2057..2074,2075..2080))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2095..2116
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2233..2250)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2404..2992)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2484..2503)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3163..3954,3955..4023))
                     /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     3786..3805
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4024..4128)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4196..4214
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4239..4454
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             4455..5258
                     /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      5369..6715
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      complement(6740..7039)
                     /label=ADH2 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 2 (ADH2) gene"
ORIGIN
        1 actttttttt atttcaaatt caagtaactg gaaggaaggc cgtataccgt tgctcattag
       61 agagtagtgt gcgtgaatga aggaaggaaa aagtttcgtg tgcttcgaga tacccctcat
      121 cagctctgga acaacgacat ctgttggtgc tgtctttgtc gttaattttt tcctttagtg
      181 tcttccatca tttttttgtc attgcggata tggtgagaca acaacggggg agagagaaaa
      241 gaaaaaaaaa gaaaagaatc cgggagttta tcattatcaa tactgccatt tcaaagaata
      301 cgtaaataat taatagtagt gattttccta actttattta gtcaaaaaat tagcctttta
      361 attctgctgt aacccgtaca tgcccaaaat agggggcggg ttacacagaa tatataacat
      421 cgtaggtgtc tgggtgaaca gtttattcct ggcatccact aaatataatg gagcccgctt
      481 tttaagctgg catccagaaa aaaaaagaat cccagcacca aaatattgtt ttcttcacca
      541 accatcagtt cataggtcca ttctcttagc gcaactacag agaacagggg cacaaacagg
      601 caaaaaacgg gcacaacctc aatggagtga tgcaacctgc ctggagtaaa tgatgacaca
      661 aggcaattga cccacgcatg tatctatctc attttcttac accttctatt accttctgct
      721 ctctctgatt tggaaaaagc tgaaaaaaaa ggttgaaacc agttccctga aattattccc
      781 ctacttgact aataagtata taaagacggt aggtattgat tgtaattctg taaatctatt
      841 tcttaaactt cttaaattct acttttatag ttagtctttt ttttagtttt aaaacaccaa
      901 gaacttagtt tcgacggatc catgcaccat caccatcacc atcaccattc cccgattttg
      961 ggttactgga aaattaaagg tttggtccaa ccgactagat tgttgttgga atacttggaa
     1021 gaaaaatacg aagaacattt gtacgaaaga gacgaaggtg acaaatggag aaacaaaaaa
     1081 tttgaattgg gtttggaatt tccgaacttg ccgtactaca ttgacggtga cgtcaaattg
     1141 actcaatcca tggcgattat tagatacatt gcggacaaac ataacatgtt gggtggttgc
     1201 ccgaaagaaa gagcggaaat ttccatgttg gaaggtgcgg tcttggacat tagatacggt
     1261 gtctccagaa ttgcgtactc caaagacttt gaaactttga aagtcgactt tttgtccaaa
     1321 ttgccggaaa tgttgaaaat gtttgaagac agattgtgcc ataaaactta cttgaacggt
     1381 gaccatgtca ctcatccgga ctttatgttg tacgacgcgt tggacgtcgt cttgtacatg
     1441 gacccgatgt gcttggacgc gtttccgaaa ttggtctgct ttaaaaaaag aattgaagcg
     1501 attccgcaaa ttgacaaata cttgaaatcc tccaaataca ttgcgtggcc gttgcaaggt
     1561 tggcaagcga cttttggtgg tggtgaccat ccgccgaaat aattctagag tcgacctgca
     1621 ggcatgcaag ctttggactt cttcgccaga ggtttggtca agtctccaat caaggttgtc
     1681 ggcttgtcta ccttgccaga aatttacgaa aagatggaaa agggtcaaat cgttggtaga
     1741 tacgttgttg acacttctaa ataagcgaat ttcttatgat ttatgatttt tattattaaa
     1801 taagttataa aaaaaataag tgtatacaaa ttttaaagtg actcttaggt tttaaaacga
     1861 aaattcttat tcttgagtaa ctctttcctg taggtcaggt tgctttctca ggtatagcat
     1921 gaggtcgctc ttattgacca cacctctacc ggcatgccgg tcgaaattcc cctaccctat
     1981 gaaaagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     2041 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     2101 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     2161 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     2221 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     2281 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     2341 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     2401 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     2461 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     2521 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     2581 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     2641 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     2701 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     2761 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     2821 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     2881 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     2941 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     3001 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     3061 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     3121 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     3181 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     3241 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     3301 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     3361 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     3421 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     3481 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     3541 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     3601 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     3661 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     3721 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     3781 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     3841 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     3901 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     3961 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     4021 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     4081 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     4141 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     4201 gcgtatcacg aggccctttc gtcttcaaga attagctttt caattcatca tttttttttt
     4261 attctttttt ttgatttcgg tttccttgaa atttttttga ttcggtaatc tccgaacaga
     4321 aggaagaacg aaggaaggag cacagactta gattggtata tatacgcata tgtagtgttg
     4381 aagaaacatg aaattgccca gtattcttaa cccaactgca cagaacaaaa acatgcagga
     4441 aacgaagata aatcatgtcg aaagctacat ataaggaacg tgctgctact catcctagtc
     4501 ctgttgctgc caagctattt aatatcatgc acgaaaagca aacaaacttg tgtgcttcat
     4561 tggatgttcg taccaccaag gaattactgg agttagttga agcattaggt cccaaaattt
     4621 gtttactaaa aacacatgtg gatatcttga ctgatttttc catggagggc acagttaagc
     4681 cgctaaaggc attatccgcc aagtacaatt ttttactctt cgaagacaga aaatttgctg
     4741 acattggtaa tacagtcaaa ttgcagtact ctgcgggtgt atacagaata gcagaatggg
     4801 cagacattac gaatgcacac ggtgtggtgg gcccaggtat tgttagcggt ttgaagcagg
     4861 cggcagaaga agtaacaaag gaacctagag gccttttgat gttagcagaa ttgtcatgca
     4921 agggctccct atctactgga gaatatacta agggtactgt tgacattgcg aagagcgaca
     4981 aagattttgt tatcggcttt attgctcaaa gagacatggg tggaagagat gaaggttacg
     5041 attggttgat tatgacaccc ggtgtgggtt tagatgacaa gggagacgca ttgggtcaac
     5101 agtatagaac cgtggatgat gtggtctcta caggatctga cattattatt gttggaagag
     5161 gactatttgc aaagggaagg gatgctaagg tagagggtga acgttacaga aaagcaggct
     5221 gggaagcata tttgagaaga tgcggccagc aaaactaaaa aactgtatta taagtaaatg
     5281 catgtatact aaactcacaa attagagctt caatttaatt atatcagtta ttacccaatt
     5341 ctcatgtttg acagcttatc atcggatcga tccaatatca aaggaaatga tagcattgaa
     5401 ggatgagact aatccaattg aggagtggca gcatatagaa cagctaaagg gtagtgctga
     5461 aggaagcata cgataccccg catggaatgg gataatatca caggaggtac tagactacct
     5521 ttcatcctac ataaatagac gcatataagt acgcatttaa gcataaacac gcactatgcc
     5581 gttcttctca tgtatatata tatacaggca acacgcagat ataggtgcga cgtgaacagt
     5641 gagctgtatg tgcgcagctc gcgttgcatt ttcggaagcg ctcgttttcg gaaacgcttt
     5701 gaagttccta ttccgaagtt cctattctct agctagaaag tataggaact tcagagcgct
     5761 tttgaaaacc aaaagcgctc tgaagacgca ctttcaaaaa accaaaaacg caccggactg
     5821 taacgagcta ctaaaatatt gcgaataccg cttccacaaa cattgctcaa aagtatctct
     5881 ttgctatata tctctgtgct atatccctat ataacctacc catccacctt tcgctccttg
     5941 aacttgcatc taaactcgac ctctacattt tttatgttta tctctagtat tactctttag
     6001 acaaaaaaat tgtagtaaga actattcata gagtgaatcg aaaacaatac gaaaatgtaa
     6061 acatttccta tacgtagtat atagagacaa aatagaagaa accgttcata attttctgac
     6121 caatgaagaa tcatcaacgc tatcactttc tgttcacaaa gtatgcgcaa tccacatcgg
     6181 tatagaatat aatcggggat gcctttatct tgaaaaaatg cacccgcagc ttcgctagta
     6241 atcagtaaac gcgggaagtg gagtcaggct ttttttatgg aagagaaaat agacaccaaa
     6301 gtagccttct tctaacctta acggacctac agtgcaaaaa gttatcaaga gactgcatta
     6361 tagagcgcac aaaggagaaa aaaagtaatc taagatgctt tgttagaaaa atagcgctct
     6421 cgggatgcat ttttgtagaa caaaaaagaa gtatagattc tttgttggta aaatagcgct
     6481 ctcgcgttgc atttctgttc tgtaaaaatg cagctcagat tctttgtttg aaaaattagc
     6541 gctctcgcgt tgcatttttg ttttacaaaa atgaagcaca gattcttcgt tggtaaaata
     6601 gcgctttcgc gttgcatttc tgttctgtaa aaatgcagct cagattcttt gtttgaaaaa
     6661 ttagcgctct cgcgttgcat ttttgttcta caaaatgaag cacagatgct tcgttccgct
     6721 cgagggcatg cgaaggaaaa tgagaaatat cgagggagac gattcagagg agcaggacaa
     6781 actataaccg actgtttgtt ggaggatgcc gtacataacg aacactgctg aagctaccat
     6841 gtctacagtt tagaggaatg ggtacaactc acaggcgagg gatggtgttc actcgtgcta
     6901 gcaaacgcgg tgggagcaaa aagtagaata ttatctttta ttcgtgaaac ttcgaacact
     6961 gtcatctaaa gatgctatat actaatatag gcatacttga taatgaaaac tataaatcgt
     7021 aaagacataa gagatccgct tatttagagt gtcgaattcg agctcggtac ccggggagtt
     7081 gattgtagct tggtatagct tgaaatattg tgcagaaaaa gaaacaagga agaaagggaa
     7141 cgagaacaat gacgaggaaa caaaagatta ataattgcag gtctatttat acttgatagc
     7201 aagacagcaa
//
