LOCUS       sequence-389770-        5482 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of N-terminally GST-tagged WzzE from Pectobacterium 
            atrosepticum in E.coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5482)
  AUTHORS   Weckener M, Woodward LS, Clarke BR, Liu H, Ward PN, Le Bas A, Bhella
            D, Whitfield C, Naismith JH
  TITLE     The lipid linked oligosaccharide polymerase Wzy and its regulating 
            co-polymerase, Wzz, from enterobacterial common antigen biosynthesis
            form a complex.
  JOURNAL   Open Biol. 2023 Mar;13(3):220373. doi: 10.1098/rsob.220373. Epub 
            2023 Mar 22.
  PUBMED    36944376
REFERENCE   2  (bases 1 to 5482)
  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: "Open 
            Biol."; date: "2023-03"; volume: "13(3):220373. doi"; pages: " 
            10.1098/rsob.220373. Epub 2023 Mar 22"
COMMENT     Sequence Label: pGST-WzzE
FEATURES             Location/Qualifiers
     source          1..5482
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(139..1221)
                     /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     1183..1202
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1222..1299)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1345..1364
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1345..1363
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1364..1388
                     /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             1432..2085
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     2043..2065
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             2113..2133
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3199..3243
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(3281..3299)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3295..3342
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(3563..4222)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4137..4156
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4223..4325)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(4717..4734)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4851..5396)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(4931..4950)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 acgatataag ttgtaattct catgttagtc atgccccgcg cccaccggaa ggagctgact
       61 gggttgaagg ctctcaaggg catcggtcga gatcccggtg cctaatgagt gagctaactt
      121 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
      181 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ccagggtggt
      241 ttttcttttc accagtgaga cgggcaacag ctgattgccc ttcaccgcct ggccctgaga
      301 gagttgcagc aagcggtcca cgctggtttg ccccagcagg cgaaaatcct gtttgatggt
      361 ggttaacggc gggatataac atgagctgtc ttcggtatcg tcgtatccca ctaccgagat
      421 gtccgcacca acgcgcagcc cggactcggt aatggcgcgc attgcgccca gcgccatctg
      481 atcgttggca accagcatcg cagtgggaac gatgccctca ttcagcattt gcatggtttg
      541 ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc gctatcggct gaatttgatt
      601 gcgagtgaga tatttatgcc agccagccag acgcagacgc gccgagacag aacttaatgg
      661 gcccgctaac agcgcgattt gctggtgacc caatgcgacc agatgctcca cgcccagtcg
      721 cgtaccgtct tcatgggaga aaataatact gttgatgggt gtctggtcag agacatcaag
      781 aaataacgcc ggaacattag tgcaggcagc ttccacagca atggcatcct ggtcatccag
      841 cggatagtta atgatcagcc cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt
      901 acaggcttcg acgccgcttc gttctaccat cgacaccacc acgctggcac ccagttgatc
      961 ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg tgcagggcca gactggaggt
     1021 ggcaacgcca atcagcaacg actgtttgcc cgccagttgt tgtgccacgc ggttgggaat
     1081 gtaattcagc tccgccatcg ccgcttccac tttttcccgc gttttcgcag aaacgtggct
     1141 ggcctggttc accacgcggg aaacggtctg ataagagaca ccggcatact ctgcgacatc
     1201 gtataacgtt actggtttca cattcaccac cctgaattga ctctcttccg ggcgctatca
     1261 tgccataccg cgaaaggttt tgcgccattc gatggtgtcc gggatctcga cgctctccct
     1321 tatgcgactc ctgcattagg aaattaatac gactcactat aggggaattg tgagcggata
     1381 acaattcccc tgtagaaata attttgttta actttaataa ggagatatac catgtcccct
     1441 atactaggtt attggaaaat taagggcctt gtgcaaccca ctcgacttct tttggaatat
     1501 cttgaagaaa aatatgaaga gcatttgtat gagcgcgatg aaggtgataa atggcgaaac
     1561 aaaaagtttg aattgggttt ggagtttccc aatcttcctt attatattga tggtgatgtt
     1621 aaattaacac agtctatggc catcatacgt tatatagctg acaagcacaa catgttgggt
     1681 ggttgtccaa aagagcgtgc agagatttca atgcttgaag gagcggtttt ggatattaga
     1741 tacggtgttt cgagaattgc atatagtaaa gactttgaaa ctctcaaagt tgattttctt
     1801 agcaagctac ctgaaatgct gaaaatgttc gaagatcgtt tatgtcataa aacatattta
     1861 aatggtgatc atgtaaccca tcctgacttc atgttgtatg acgctcttga tgttgtttta
     1921 tacatggacc caatgtgcct ggatgcgttc ccaaaattag tttgttttaa aaaacgtatt
     1981 gaagctatcc cacaaattga taagtacttg aaatccagca agtatatagc atggcctttg
     2041 cagggctggc aagccacgtt tggtggtggc gaccatcctc caaaatcgga tgattacgac
     2101 atcccaacga ccgaaaacct gtattttcag ggcgccatgg tgaaatcaga gaacttgtct
     2161 accgggaatg cgttgattga taacgaactg gatatccgtg gtttattccg cctgctgtgg
     2221 cgtggaaagg tgtggccgat agcaatcggc ctgctgtttg ccgtcgtggc attaggctat
     2281 tcctatcttg ttaaacagga atggggtgca acatcgatta ccgacaagcc gacggttaat
     2341 atgttgggcg gctattactc gcagcagcaa ttcctacgta acctcgacgc tcgatcgttt
     2401 tcctctcctc agccagaaca accgtctatt tcggctggtg cttatgatga attcatcatg
     2461 cagttagcgg cttacgatac ccgccgcgat ttctggttgc agactgacta ttataagcaa
     2521 cgtctggaag gtgatgaaaa ggctgatgca gccttgctgg atgagctggt caacaacatt
     2581 cagtttacgg cgcgtgatga cgggaaaaaa accaatgact ccgtgaagct ggtggcggaa
     2641 acctcctcag attccaacac gttgcttcgt cagtatgtgg tttttgccag tcagcgtgca
     2701 gccaaccact tgaatgaaga aatcaaggga gcctgggctg ccagaaccat tttcatgaag
     2761 tcgcagatta agcgtcagga agcggtagca aaagcgattt acgatcgtga agtccgcagc
     2821 gttgaactgg cgctgaaaat cgcacaacag caaggtatta gccgtagcca gacggatacg
     2881 ccagcagatg aaatccccgc atcggaaatg ttcctgctcg gcaggccgat gcttcaagcc
     2941 cgactggaaa cgttgcagac cagcggcccg cattatgaac tggattacga tcaaaatcgc
     3001 gcgatgttgg cgacgctgaa cgttggccca acgcttgagg ccagtttcca gacgtatcgt
     3061 tatttgcgta caccggaaga accggtgaaa cgcgacagcc cgcgccgggc attcctgatg
     3121 gtgatgtggg gtgcgattgg tgtgctagtg ggagcgggtg ttgccctggc tcgccgtctg
     3181 cgttagctcg agtctggtaa agaaaccgct gctgcgaaat ttgaacgcca gcacatggac
     3241 tcgtctacta gcgcagctta attaacctag gctgctgcca ccgctgagca ataactagca
     3301 taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaacc tcaggcattt
     3361 gagaagcaca cggtcacact gcttccggta gtcaataaac cggtaaacca gcaatagaca
     3421 taagcggcta tttaacgacc ctgccctgaa ccgacgaccg ggtcgaattt gctttcgaat
     3481 ttctgccatt catccgctta ttatcactta ttcaggcgta gcaaccaggc gtttaagggc
     3541 accaataact gccttaaaaa aattacgccc cgccctgcca ctcatcgcag tactgttgta
     3601 attcattaag cattctgccg acatggaagc catcacaaac ggcatgatga acctgaatcg
     3661 ccagcggcat cagcaccttg tcgccttgcg tataatattt gcccatagtg aaaacggggg
     3721 cgaagaagtt gtccatattg gccacgttta aatcaaaact ggtgaaactc acccagggat
     3781 tggctgagac gaaaaacata ttctcaataa accctttagg gaaataggcc aggttttcac
     3841 cgtaacacgc cacatcttgc gaatatatgt gtagaaactg ccggaaatcg tcgtggtatt
     3901 cactccagag cgatgaaaac gtttcagttt gctcatggaa aacggtgtaa caagggtgaa
     3961 cactatccca tatcaccagc tcaccgtctt tcattgccat acggaactcc ggatgagcat
     4021 tcatcaggcg ggcaagaatg tgaataaagg ccggataaaa cttgtgctta tttttcttta
     4081 cggtctttaa aaaggccgta atatccagct gaacggtctg gttataggta cattgagcaa
     4141 ctgactgaaa tgcctcaaaa tgttctttac gatgccattg ggatatatca acggtggtat
     4201 atccagtgat ttttttctcc attttagctt ccttagctcc tgaaaatctc gataactcaa
     4261 aaaatacgcc cggtagtgat cttatttcat tatggtgaaa gttggaacct cttacgtgcc
     4321 gatcaacgtc tcattttcgc caaaagttgg cccagggctt cccggtatca acagggacac
     4381 caggatttat ttattctgcg aagtgatctt ccgtcacagg tatttattcg gcgcaaagtg
     4441 cgtcgggtga tgctgccaac ttactgattt agtgtatgat ggtgtttttg aggtgctcca
     4501 gtggcttctg tttctatcag ctgtccctcc tgttcagcta ctgacggggt ggtgcgtaac
     4561 ggcaaaagca ccgccggaca tcagcgctag cggagtgtat actggcttac tatgttggca
     4621 ctgatgaggg tgtcagtgaa gtgcttcatg tggcaggaga aaaaaggctg caccggtgcg
     4681 tcagcagaat atgtgataca ggatatattc cgcttcctcg ctcactgact cgctacgctc
     4741 ggtcgttcga ctgcggcgag cggaaatggc ttacgaacgg ggcggagatt tcctggaaga
     4801 tgccaggaag atacttaaca gggaagtgag agggccgcgg caaagccgtt tttccatagg
     4861 ctccgccccc ctgacaagca tcacgaaatc tgacgctcaa atcagtggtg gcgaaacccg
     4921 acaggactat aaagatacca ggcgtttccc cctggcggct ccctcgtgcg ctctcctgtt
     4981 cctgcctttc ggtttaccgg tgtcattccg ctgttatggc cgcgtttgtc tcattccacg
     5041 cctgacactc agttccgggt aggcagttcg ctccaagctg gactgtatgc acgaaccccc
     5101 cgttcagtcc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggaaag
     5161 acatgcaaaa gcaccactgg cagcagccac tggtaattga tttagaggag ttagtcttga
     5221 agtcatgcgc cggttaaggc taaactgaaa ggacaagttt tggtgactgc gctcctccaa
     5281 gccagttacc tcggttcaaa gagttggtag ctcagagaac cttcgaaaaa ccgccctgca
     5341 aggcggtttt ttcgttttca gagcaagaga ttacgcgcag accaaaacga tctcaagaag
     5401 atcatcttat taatcagata aaatatttct agatttcagt gcaatttatc tcttcaaatg
     5461 tagcacctga agtcagcccc at
//
