LOCUS       sequence-425614-        5485 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Xylose reductase from Hypocrea jecorina fused with GST-tagged and 
            codon optimized for expression in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5485)
  AUTHORS   Jaroensuk J, Sutthaphirom C, Phonbuppha J, Chinantuya W, Kesornpun 
            C, Akeratchatapan N, Kittipanukul N, Phatinuwat K, Atichartpongkul 
            S, Fuangthong M, Pongtharangkul T, Hollmann F, Chaiyen P
  TITLE     A versatile in situ cofactor enhancing system for meeting cellular 
            demands for engineered metabolic pathways.
  JOURNAL   J Biol Chem. 2023 Dec 28:105598. doi: 10.1016/j.jbc.2023.105598.
  PUBMED    38159859
REFERENCE   2  (bases 1 to 5485)
  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: "J Biol 
            Chem. 2023 Dec 28:105598. doi: 10.1016/j.jbc.2023.105598."
COMMENT     Sequence Label: pRSFDuet-gst-xr
FEATURES             Location/Qualifiers
     source          1..5485
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    3..27
                     /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)."
     CDS             83..100
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        214..232
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    233..257
                     /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)."
     CDS             299..952
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     910..932
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             959..976
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             2022..2066
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(2104..2122)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2118..2165
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2395..3210)
                     /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(2395..3210)
                     /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     3118..3137
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(3211..3302)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3318..4067)
                     /direction=LEFT
                     /label=RSF ori
                     /note="Plasmids containing the RSF 1030 origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     primer_bind     complement(3328..3345)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3568..3587)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4263..5345)
                     /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     5307..5326
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5346..5423)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        join(5469..5485,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag
       61 gagatatacc atgggcagca gccatcacca tcatcaccac agccaggatc cgaattcgag
      121 ctcggcgcgc ctgcaggtcg acaagcttgc ggccgcataa tgcttaagtc gaacagaaag
      181 taatcgtatt gtacacggcc gcataatcga aattaatacg actcactata ggggaattgt
      241 gagcggataa caattcccca tcttagtata ttagttaagt ataagaagga gatatacaat
      301 gtcccctata ctaggttatt ggaaaattaa gggccttgtg caacccactc gacttctttt
      361 ggaatatctt gaagaaaaat atgaagagca tttgtatgag cgcgatgaag gtgataaatg
      421 gcgaaacaaa aagtttgaat tgggtttgga gtttcccaat cttccttatt atattgatgg
      481 tgatgttaaa ttaacacagt ctatggccat catacgttat atagctgaca agcacaacat
      541 gttgggtggt tgtccaaaag agcgtgcaga gatttcaatg cttgaaggag cggttttgga
      601 tattagatac ggtgtttcga gaattgcata tagtaaagac tttgaaactc tcaaagttga
      661 ttttcttagc aagctacctg aaatgctgaa aatgttcgaa gatcgtttat gtcataaaac
      721 atatttaaat ggtgatcatg taacccatcc tgacttcatg ttgtatgacg ctcttgatgt
      781 tgttttatac atggacccaa tgtgcctgga tgcgttccca aaattagttt gttttaaaaa
      841 acgtattgaa gctatcccac aaattgataa gtacttgaaa tccagcaagt atatagcatg
      901 gcctttgcag ggctggcaag ccacgtttgg tggtggcgac catcctccaa aatcggatct
      961 ggttccgcgt ggatccatgg cgagcccgac cctgaagctg aatagcggtt acgatatgcc
     1021 gcaagttggt tttggcctgt ggaaggttga taatgcggtg tgcgcggaca ccgtgtacaa
     1081 cgcgatcaag gcgggttatc gtctgttcga cggtgcgtgc gattacggca acgagaaaga
     1141 atgcggcgag ggcgttgcgc gtgcgatcaa ggacggcctg gtgaaacgtg aagacctgtt
     1201 cattgttagc aagctgtggc agacctttca cgacgaggat aaagtggaac cgatcacccg
     1261 tcgtcagctg gcggattggc aaattgacta cttcgacctg ttcctggttc acttcccggc
     1321 ggcgctggaa tatgtggacc cgagcgttcg ttacccgccg ggttggtttt atgatggcaa
     1381 gagcgaggtg cgctggagca aaaccaccac cctgcagcaa acctggggtg cgatggaacg
     1441 tctggttgac aagggtctgg cgcgtagcat tggcgtgagc aactatcagg cgcaaagcgt
     1501 ttacgatgcg ctgatctatg cgcgtattaa accggcgacc ctgcagattg agcaccaccc
     1561 gtacctgcag caaccggacc tggtgagcct ggcgcaaacc gaaggtattg tggttaccgc
     1621 gtatagcagc ttcggtccga ccggctttat ggagctggat atgccgcgtg cgaagagcgt
     1681 ggcgccgctg atggacagcc cggttatcaa ggcgctggcg gataaacatc gtcgtacccc
     1741 ggcgcaggtt ctgctgcgtt gggcgaccca acgtggcatc gcggtgattc cgaagaccag
     1801 ccgtccggaa gtgatggcgc aaaacctgga caacaccagc tttgacctgg atagcgaaga
     1861 tctggcgaaa attgcggaca tggatctgaa catccgtttc aataagccga ccaactactt
     1921 tagcgcgaat aagctgtacc tgtttggtta agaatttggc agatctcaat tggatatcgg
     1981 ccggccacgc gatcgctgac gtcggtaccc tcgagtctgg taaagaaacc gctgctgcga
     2041 aatttgaacg ccagcacatg gactcgtcta ctagcgcagc ttaattaacc taggctgctg
     2101 ccaccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt
     2161 ttttgctgaa acctcaggca tttgagaagc acacggtcac actgcttccg gtagtcaata
     2221 aaccggtaaa ccagcaatag acataagcgg ctatttaacg accctgccct gaaccgacga
     2281 caagctgacg accgggtctc cgcaagtggc acttttcggg gaaatgtgcg cggaacccct
     2341 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgaatta attcttagaa
     2401 aaactcatcg agcatcaaat gaaactgcaa tttattcata tcaggattat caataccata
     2461 tttttgaaaa agccgtttct gtaatgaagg agaaaactca ccgaggcagt tccataggat
     2521 ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca acatcaatac aacctattaa
     2581 tttcccctcg tcaaaaataa ggttatcaag tgagaaatca ccatgagtga cgactgaatc
     2641 cggtgagaat ggcaaaagtt tatgcatttc tttccagact tgttcaacag gccagccatt
     2701 acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg
     2761 agcgagacga aatacgcggt cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa
     2821 ccggcgcagg aacactgcca gcgcatcaac aatattttca cctgaatcag gatattcttc
     2881 taatacctgg aatgctgttt tcccggggat cgcagtggtg agtaaccatg catcatcagg
     2941 agtacggata aaatgcttga tggtcggaag aggcataaat tccgtcagcc agtttagtct
     3001 gaccatctca tctgtaacat cattggcaac gctacctttg ccatgtttca gaaacaactc
     3061 tggcgcatcg ggcttcccat acaatcgata gattgtcgca cctgattgcc cgacattatc
     3121 gcgagcccat ttatacccat ataaatcagc atccatgttg gaatttaatc gcggcctaga
     3181 gcaagacgtt tcccgttgaa tatggctcat actcttcctt tttcaatatt attgaagcat
     3241 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     3301 aataggcatg cagcgctctt ccgcttcctc gctcactgac tcgctacgct cggtcgttcg
     3361 actgcggcga gcggtgtcag ctcactcaaa agcggtaata cggttatcca cagaatcagg
     3421 ggataaagcc ggaaagaaca tgtgagcaaa aagcaaagca ccggaagaag ccaacgccgc
     3481 aggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagcc
     3541 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     3601 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     3661 cgggaagcgt ggcgctttct catagctcac gctgttggta tctcagttcg gtgtaggtcg
     3721 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     3781 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     3841 ccattggtaa ctgatttaga ggactttgtc ttgaagttat gcacctgtta aggctaaact
     3901 gaaagaacag attttggtga gtgcggtcct ccaacccact taccttggtt caaagagttg
     3961 gtagctcagc gaaccttgag aaaaccaccg ttggtagcgg tggtttttct ttatttatga
     4021 gatgatgaat caatcggtct atcaagtcaa cgaacagcta ttccgttact ctagatttca
     4081 gtgcaattta tctcttcaaa tgtagcacct gaagtcagcc ccatacgata taagttgtaa
     4141 ttctcatgtt agtcatgccc cgcgcccacc ggaaggagct gactgggttg aaggctctca
     4201 agggcatcgg tcgagatccc ggtgcctaat gagtgagcta acttacatta attgcgttgc
     4261 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     4321 aacgcgcggg gagaggcggt ttgcgtattg ggcgccaggg tggtttttct tttcaccagt
     4381 gagacgggca acagctgatt gcccttcacc gcctggccct gagagagttg cagcaagcgg
     4441 tccacgctgg tttgccccag caggcgaaaa tcctgtttga tggtggttaa cggcgggata
     4501 taacatgagc tgtcttcggt atcgtcgtat cccactaccg agatgtccgc accaacgcgc
     4561 agcccggact cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc
     4621 atcgcagtgg gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa accggacatg
     4681 gcactccagt cgccttcccg ttccgctatc ggctgaattt gattgcgagt gagatattta
     4741 tgccagccag ccagacgcag acgcgccgag acagaactta atgggcccgc taacagcgcg
     4801 atttgctggt gacccaatgc gaccagatgc tccacgccca gtcgcgtacc gtcttcatgg
     4861 gagaaaataa tactgttgat gggtgtctgg tcagagacat caagaaataa cgccggaaca
     4921 ttagtgcagg cagcttccac agcaatggca tcctggtcat ccagcggata gttaatgatc
     4981 agcccactga cgcgttgcgc gagaagattg tgcaccgccg ctttacaggc ttcgacgccg
     5041 cttcgttcta ccatcgacac caccacgctg gcacccagtt gatcggcgcg agatttaatc
     5101 gccgcgacaa tttgcgacgg cgcgtgcagg gccagactgg aggtggcaac gccaatcagc
     5161 aacgactgtt tgcccgccag ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc
     5221 atcgccgctt ccactttttc ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg
     5281 cgggaaacgg tctgataaga gacaccggca tactctgcga catcgtataa cgttactggt
     5341 ttcacattca ccaccctgaa ttgactctct tccgggcgct atcatgccat accgcgaaag
     5401 gttttgcgcc attcgatggt gtccgggatc tcgacgctct cccttatgcg actcctgcat
     5461 taggaaatta atacgactca ctata
//
