LOCUS       sequence-206044-        5931 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for bacterial expression of a His-tagged A180Q mutant of 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: pTH860-gst-optec A180Q
FEATURES             Location/Qualifiers
     source          1..5931
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     90..109
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     193..212
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        193..211
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    212..236
                     /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             251..273
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             286..309
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             988..1005
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1058..1076)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1072..1119
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1156..1611
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1243..1262)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1453..1474
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1703..2518)
                     /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(1703..2518)
                     /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     2426..2445
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2510..2529
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2640..3228
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3129..3148
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3382..3399
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3414..3553
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3479..3498)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3639..3661
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3655..3846)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4655..5737)
                     /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     5699..5718
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5738..5815)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 aacagtcccc cggccacggg gcctgccacc atacccacgc cgaaacaagc gctcatgagc
       61 ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg cgatataggc gccagcaacc
      121 gcacctgtgg cgccggtgat gccggccacg atgcgtccgg cgtagaggat cgagatctcg
      181 atcccgcgaa attaatacga ctcactatag gggaattgtg agcggataac aattcccctc
      241 tagaaataat tttgtttaac tttaagaagg agatatacca tgggccacca tcaccatcac
      301 catcaccata gcccgattct gggctattgg aaaattaaag gcctggtgca gccgacccgc
      361 ctgctgctgg aatatctgga agaaaaatat gaagaacatc tgtatgaacg cgatgaaggc
      421 gataaatggc gcaacaaaaa atttgaactg ggcctggaat ttccgaacct gccgtattat
      481 attgatggcg atgtgaaact gacccagagc atggcgatta ttcgctatat tgcggataaa
      541 cataacatgc tgggcggctg cccgaaagaa cgcgcggaaa ttagcatgct ggaaggcgcg
      601 gtgctggata ttcgctatgg cgtgagccgc attgcgtata gcaaagattt tgaaaccctg
      661 aaagtggatt ttctgagcaa actgccggaa atgctgaaaa tgtttgaaga tcgcctgtgc
      721 cataaaacct atctgaacgg cgatcatgtg acccatccgg attttatgct gtatgatgcg
      781 ctggatgtgg tgctgtatat ggatccgatg tgcctggatc agtttccgaa actggtgtgc
      841 tttaaaaagc gcattgaagc gattccgcag attgataaat atctgaaaag cagcaaatat
      901 attgcgtggc cgctgcaggg ctggcaggcg acctttggcg gcggcgatca tccgccgaaa
      961 taataaaagc ttgcggccgc actcgagcac caccaccacc accactgaga tccggctgct
     1021 aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa
     1081 ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc
     1141 ggattggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     1201 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     1261 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     1321 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     1381 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     1441 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     1501 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     1561 tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt taggtggcac
     1621 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     1681 gtatccgctc atgaattaat tcttagaaaa actcatcgag catcaaatga aactgcaatt
     1741 tattcatatc aggattatca ataccatatt tttgaaaaag ccgtttctgt aatgaaggag
     1801 aaaactcacc gaggcagttc cataggatgg caagatcctg gtatcggtct gcgattccga
     1861 ctcgtccaac atcaatacaa cctattaatt tcccctcgtc aaaaataagg ttatcaagtg
     1921 agaaatcacc atgagtgacg actgaatccg gtgagaatgg caaaagttta tgcatttctt
     1981 tccagacttg ttcaacaggc cagccattac gctcgtcatc aaaatcactc gcatcaacca
     2041 aaccgttatt cattcgtgat tgcgcctgag cgagacgaaa tacgcgatcg ctgttaaaag
     2101 gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa
     2161 tattttcacc tgaatcagga tattcttcta atacctggaa tgctgttttc ccggggatcg
     2221 cagtggtgag taaccatgca tcatcaggag tacggataaa atgcttgatg gtcggaagag
     2281 gcataaattc cgtcagccag tttagtctga ccatctcatc tgtaacatca ttggcaacgc
     2341 tacctttgcc atgtttcaga aacaactctg gcgcatcggg cttcccatac aatcgataga
     2401 ttgtcgcacc tgattgcccg acattatcgc gagcccattt atacccatat aaatcagcat
     2461 ccatgttgga atttaatcgc ggcctagagc aagacgtttc ccgttgaata tggctcataa
     2521 caccccttgt attactgttt atgtaagcag acagttttat tgttcatgac caaaatccct
     2581 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     2641 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     2701 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     2761 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
     2821 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     2881 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     2941 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     3001 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     3061 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     3121 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     3181 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     3241 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     3301 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     3361 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg
     3421 cggtattttc tccttacgca tctgtgcggt atttcacacc gcaatggtgc actctcagta
     3481 caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg
     3541 ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     3601 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     3661 gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc
     3721 gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag
     3781 aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt
     3841 ggtcactgat gcctccgtgt aagggggatt tctgttcatg ggggtaatga taccgatgaa
     3901 acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg
     3961 ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg
     4021 tcaatgccag cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc
     4081 tgcgatgcag atccggaaca taatggtgca gggcgctgac ttccgcgttt ccagacttta
     4141 cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca
     4201 gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc
     4261 ccgccagcct agccgggtcc tcaacgacag gagcacgatc atgcgcaccc gtggggccgc
     4321 catgccggcg ataatggcct gcttctcgcc gaaacgtttg gtggcgggac cagtgacgaa
     4381 ggcttgagcg agggcgtgca agattccgaa taccgcaagc gacaggccga tcatcgtcgc
     4441 gctccagcga aagcggtcct cgccgaaaat gacccagagc gctgccggca cctgtcctac
     4501 gagttgcatg ataaagaaga cagtcataag tgcggcgacg atagtcatgc cccgcgccca
     4561 ccggaaggag ctgactgggt tgaaggctct caagggcatc ggtcgagatc ccggtgccta
     4621 atgagtgagc taacttacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     4681 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     4741 tgggcgccag ggtggttttt cttttcacca gtgagacggg caacagctga ttgcccttca
     4801 ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa
     4861 aatcctgttt gatggtggtt aacggcggga tataacatga gctgtcttcg gtatcgtcgt
     4921 atcccactac cgagatatcc gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg
     4981 cgcccagcgc catctgatcg ttggcaacca gcatcgcagt gggaacgatg ccctcattca
     5041 gcatttgcat ggtttgttga aaaccggaca tggcactcca gtcgccttcc cgttccgcta
     5101 tcggctgaat ttgattgcga gtgagatatt tatgccagcc agccagacgc agacgcgccg
     5161 agacagaact taatgggccc gctaacagcg cgatttgctg gtgacccaat gcgaccagat
     5221 gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat aatactgttg atgggtgtct
     5281 ggtcagagac atcaagaaat aacgccggaa cattagtgca ggcagcttcc acagcaatgg
     5341 catcctggtc atccagcgga tagttaatga tcagcccact gacgcgttgc gcgagaagat
     5401 tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc taccatcgac accaccacgc
     5461 tggcacccag ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca
     5521 gggccagact ggaggtggca acgccaatca gcaacgactg tttgcccgcc agttgttgtg
     5581 ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc ttccactttt tcccgcgttt
     5641 tcgcagaaac gtggctggcc tggttcacca cgcgggaaac ggtctgataa gagacaccgg
     5701 catactctgc gacatcgtat aacgttactg gtttcacatt caccaccctg aattgactct
     5761 cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg gtgtccggga
     5821 tctcgacgct ctcccttatg cgactcctgc attaggaagc agcccagtag taggttgagg
     5881 ccgttgagca ccgccgccgc aaggaatggt gcatgcaagg agatggcgcc c
//
