LOCUS       sequence-206038-        5931 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for bacterial expression of His-tagged GST encoded by 
            preferred codons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5931)
  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 5931)
  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: pTH849-gst-optec
FEATURES             Location/Qualifiers
     source          1..5931
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     15..34
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     118..137
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        118..136
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    137..161
                     /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)."
     RBS             176..198
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             211..234
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             235..888
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="SPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKKF
                     ELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRYG
                     VSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVLY
                     MDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     CDS             913..930
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(983..1001)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      997..1044
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1081..1536
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1168..1187)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1378..1399
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1628..2443)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(1628..2443)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2351..2370
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2435..2454
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2565..3153
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3054..3073
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3307..3324
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3339..3478
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3404..3423)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3564..3586
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3580..3771)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4580..5662)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     5624..5643
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5663..5740)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 cgatcttccc catcggtgat gtcggcgata taggcgccag caaccgcacc tgtggcgccg
       61 gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga tctcgatccc gcgaaattaa
      121 tacgactcac tataggggaa ttgtgagcgg ataacaattc ccctctagaa ataattttgt
      181 ttaactttaa gaaggagata taccatgggc caccatcacc atcaccatca ccatagcccg
      241 attctgggct attggaaaat taaaggcctg gtgcagccga cccgcctgct gctggaatat
      301 ctggaagaaa aatatgaaga acatctgtat gaacgcgatg aaggcgataa atggcgcaac
      361 aaaaaatttg aactgggcct ggaatttccg aacctgccgt attatattga tggcgatgtg
      421 aaactgaccc agagcatggc gattattcgc tatattgcgg ataaacataa catgctgggc
      481 ggctgcccga aagaacgcgc ggaaattagc atgctggaag gcgcggtgct ggatattcgc
      541 tatggcgtga gccgcattgc gtatagcaaa gattttgaaa ccctgaaagt ggattttctg
      601 agcaaactgc cggaaatgct gaaaatgttt gaagatcgcc tgtgccataa aacctatctg
      661 aacggcgatc atgtgaccca tccggatttt atgctgtatg atgcgctgga tgtggtgctg
      721 tatatggatc cgatgtgcct ggatgcgttt ccgaaactgg tgtgctttaa aaaacgcatt
      781 gaagcgattc cgcagattga taaatatctg aaaagcagca aatatattgc gtggccgctg
      841 cagggctggc aggcgacctt tggcggcggc gatcatccgc cgaaataata aaagcttgcg
      901 gccgcactcg agcaccacca ccaccaccac tgagatccgg ctgctaacaa agcccgaaag
      961 gaagctgagt tggctgctgc caccgctgag caataactag cataacccct tggggcctct
     1021 aaacgggtct tgaggggttt tttgctgaaa ggaggaacta tatccggatt ggcgaatggg
     1081 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     1141 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     1201 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     1261 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     1321 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     1381 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     1441 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     1501 acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt
     1561 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgaa
     1621 ttaattctta gaaaaactca tcgagcatca aatgaaactg caatttattc atatcaggat
     1681 tatcaatacc atatttttga aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc
     1741 agttccatag gatggcaaga tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa
     1801 tacaacctat taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag
     1861 tgacgactga atccggtgag aatggcaaaa gtttatgcat ttctttccag acttgttcaa
     1921 caggccagcc attacgctcg tcatcaaaat cactcgcatc aaccaaaccg ttattcattc
     1981 gtgattgcgc ctgagcgaga cgaaatacgc gatcgctgtt aaaaggacaa ttacaaacag
     2041 gaatcgaatg caaccggcgc aggaacactg ccagcgcatc aacaatattt tcacctgaat
     2101 caggatattc ttctaatacc tggaatgctg ttttcccggg gatcgcagtg gtgagtaacc
     2161 atgcatcatc aggagtacgg ataaaatgct tgatggtcgg aagaggcata aattccgtca
     2221 gccagtttag tctgaccatc tcatctgtaa catcattggc aacgctacct ttgccatgtt
     2281 tcagaaacaa ctctggcgca tcgggcttcc catacaatcg atagattgtc gcacctgatt
     2341 gcccgacatt atcgcgagcc catttatacc catataaatc agcatccatg ttggaattta
     2401 atcgcggcct agagcaagac gtttcccgtt gaatatggct cataacaccc cttgtattac
     2461 tgtttatgta agcagacagt tttattgttc atgaccaaaa tcccttaacg tgagttttcg
     2521 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     2581 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     2641 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     2701 ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     2761 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     2821 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     2881 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     2941 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     3001 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     3061 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     3121 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     3181 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     3241 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     3301 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt
     3361 acgcatctgt gcggtatttc acaccgcaat ggtgcactct cagtacaatc tgctctgatg
     3421 ccgcatagtt aagccagtat acactccgct atcgctacgt gactgggtca tggctgcgcc
     3481 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     3541 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     3601 accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca
     3661 gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg ttaatgtctg
     3721 gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca ctgatgcctc
     3781 cgtgtaaggg ggatttctgt tcatgggggt aatgataccg atgaaacgag agaggatgct
     3841 cacgatacgg gttactgatg atgaacatgc ccggttactg gaacgttgtg agggtaaaca
     3901 actggcggta tggatgcggc gggaccagag aaaaatcact cagggtcaat gccagcgctt
     3961 cgttaataca gatgtaggtg ttccacaggg tagccagcag catcctgcga tgcagatccg
     4021 gaacataatg gtgcagggcg ctgacttccg cgtttccaga ctttacgaaa cacggaaacc
     4081 gaagaccatt catgttgttg ctcaggtcgc agacgttttg cagcagcagt cgcttcacgt
     4141 tcgctcgcgt atcggtgatt cattctgcta accagtaagg caaccccgcc agcctagccg
     4201 ggtcctcaac gacaggagca cgatcatgcg cacccgtggg gccgccatgc cggcgataat
     4261 ggcctgcttc tcgccgaaac gtttggtggc gggaccagtg acgaaggctt gagcgagggc
     4321 gtgcaagatt ccgaataccg caagcgacag gccgatcatc gtcgcgctcc agcgaaagcg
     4381 gtcctcgccg aaaatgaccc agagcgctgc cggcacctgt cctacgagtt gcatgataaa
     4441 gaagacagtc ataagtgcgg cgacgatagt catgccccgc gcccaccgga aggagctgac
     4501 tgggttgaag gctctcaagg gcatcggtcg agatcccggt gcctaatgag tgagctaact
     4561 tacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     4621 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gccagggtgg
     4681 tttttctttt caccagtgag acgggcaaca gctgattgcc cttcaccgcc tggccctgag
     4741 agagttgcag caagcggtcc acgctggttt gccccagcag gcgaaaatcc tgtttgatgg
     4801 tggttaacgg cgggatataa catgagctgt cttcggtatc gtcgtatccc actaccgaga
     4861 tatccgcacc aacgcgcagc ccggactcgg taatggcgcg cattgcgccc agcgccatct
     4921 gatcgttggc aaccagcatc gcagtgggaa cgatgccctc attcagcatt tgcatggttt
     4981 gttgaaaacc ggacatggca ctccagtcgc cttcccgttc cgctatcggc tgaatttgat
     5041 tgcgagtgag atatttatgc cagccagcca gacgcagacg cgccgagaca gaacttaatg
     5101 ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc acgcccagtc
     5161 gcgtaccgtc ttcatgggag aaaataatac tgttgatggg tgtctggtca gagacatcaa
     5221 gaaataacgc cggaacatta gtgcaggcag cttccacagc aatggcatcc tggtcatcca
     5281 gcggatagtt aatgatcagc ccactgacgc gttgcgcgag aagattgtgc accgccgctt
     5341 tacaggcttc gacgccgctt cgttctacca tcgacaccac cacgctggca cccagttgat
     5401 cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc agactggagg
     5461 tggcaacgcc aatcagcaac gactgtttgc ccgccagttg ttgtgccacg cggttgggaa
     5521 tgtaattcag ctccgccatc gccgcttcca ctttttcccg cgttttcgca gaaacgtggc
     5581 tggcctggtt caccacgcgg gaaacggtct gataagagac accggcatac tctgcgacat
     5641 cgtataacgt tactggtttc acattcacca ccctgaattg actctcttcc gggcgctatc
     5701 atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc cgggatctcg acgctctccc
     5761 ttatgcgact cctgcattag gaagcagccc agtagtaggt tgaggccgtt gagcaccgcc
     5821 gccgcaagga atggtgcatg caaggagatg gcgcccaaca gtcccccggc cacggggcct
     5881 gccaccatac ccacgccgaa acaagcgctc atgagcccga agtggcgagc c
//
