LOCUS       sequence-389760-        5739 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of C-terminally GST-distrep tag II-tagged WzyE from 
            Pectobacterium atrosepticum in E.coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5739)
  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 5739)
  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: pWzyE-GST-distrepII
FEATURES             Location/Qualifiers
     source          1..5739
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      90..678
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     579..598
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     832..849
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    864..1003
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(929..948)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1089..1111
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1105..1296)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     1805..1824
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1805..1823
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1855..1877
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3294..3944
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="SPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKKF
                     ELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRYG
                     VSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVLY
                     MDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     3902..3924
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             3954..3974
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3978..4001
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             4038..4061
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(4114..4132)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4128..4175
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      4212..4667
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4299..4318)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4509..4530
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4693..4797
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4798..4866,4867..5658)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5016..5035)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
       61 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
      121 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
      181 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt
      241 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
      301 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
      361 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
      421 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
      481 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
      541 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
      601 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
      661 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
      721 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
      781 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
      841 gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt
      901 atttcacacc gcaatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
      961 agtatacact ccgctatcgc tacgtgactg ggtcatggct gcgccccgac acccgccaac
     1021 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     1081 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     1141 gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc
     1201 atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc tgataaagcg
     1261 ggccatgtta agggcggttt tttcctgttt ggtcactgat gcctccgtgt aagggggatt
     1321 tctgttcatg ggggtaatga taccgatgaa acgagagagg atgctcacga tacgggttac
     1381 tgatgatgaa catgcccggt tactggaacg ttgtgagggt aaacaactgg cggtatggat
     1441 gcggcgggac cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta atacagatgt
     1501 aggtgttcca cagggtagcc agcagcatcc tgcgatgcag atccggaaca taatggtgca
     1561 gggcgctgac ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga ccattcatgt
     1621 tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct cgcgtatcgg
     1681 tgattcattc tgctaaccag taaggcaacc ccgccagcct agccgggtcc tcaacgacag
     1741 gagcacgatc atgcgcaccc gtggccagga cccaacgctg cccgagatct cgatcccgcg
     1801 aaattaatac gactcactat agggagacca caacggtttc cctctagaaa taattttgtt
     1861 taactttaag aaggagatat accatggcgc ttgggcaatt tggagggctg tttgtcgtct
     1921 acctgatttc agtcatcttc atcctgagct tgacctggat ggagtttcgg cgcgtacgtt
     1981 ttaactttaa cgtgctgttc tcgctgctct atttattgac gttctatttt ggttttccct
     2041 ttacctgcgt gctggtgttc cggtttggcg ttgaagtcgt ccccgtgcag tatctgcttc
     2101 aggcgatgtt gtccgcaacg gcgttctatg ctgtctatta cgtcagctac aagacgcgac
     2161 tacgccagaa aacctccgtg ccgcgtgctc cactcctgac ggtgaaccga gttgaagcta
     2221 atctgacctg gctgctgctg gcgttgattg cggtagccac cgtcggtatt ttcttcctca
     2281 acaatggctt cctgctgttt aagctgcgct cttacagcca gatattctct agcgatgtgt
     2341 ccggtgtggc gttgaaacgc tttttctact tcttcattcc ggcaatgctg gtggtgtatt
     2401 tcctgcgcca gacgcagcga gcgtggctac tgttcctcat cggaacggtc gcatttggaa
     2461 tgctgacata tgtgattgtc ggcgggacgc gcgcgaatct gatcattgcc ttcgccctat
     2521 ttctgtttat tggcattgtg cgtggttgga ttacgctctg gatgttggtc gctgctggga
     2581 tatttggtat tgtcgggatg ttctggctgg cgctgaagcg ctatgggctg gacgtcagcg
     2641 gtgattacgc attttatacc ttcctctatt taactcgtga caccttctcg ccctgggaga
     2701 atctggcgct gctgtggcag aattatgaca agattgagtt tcaggggctg gcaccgattg
     2761 ctcgtgattt ctacgtcttt attccctcct ggctatggcc tgaccgtccg aatctggtgt
     2821 taaacagcgc gaactatttc acctgggaag tgttgaataa ccactctggg ctggcgatat
     2881 cccctacgct gttaggctcg ctggtggtga tgggcggcgt gctatttatc ccgttgggtg
     2941 cgattgcggt agggctggtt atcaagtggt tcgactgggt gtacgagtta gggaagaacg
     3001 acagcaatcg ctataaggcc gccattttgc aggcgttctg cttcggtgcg gtgttcaata
     3061 ttattgtgct aacgcgcgaa ggcgtcgatt cctttgtttc acgtgtggtg tttttctgtc
     3121 tgatatttgg cctgtgcctg ctcgttgcca aactgcttta ctggctgctg gaaagcgccg
     3181 gactcatccg gcagcgtctg aggcgcatgc gcgccacgcc gctcgcgccg acacccaata
     3241 ccgtacagga tccggtgata aaggagcaga tactcgaggg cggtggcggt tcgtccccta
     3301 tactaggtta ttggaaaatt aagggccttg tgcaacccac tcgacttctt ttggaatatc
     3361 ttgaagaaaa atatgaagag catttgtatg agcgcgatga aggtgataaa tggcgaaaca
     3421 aaaagtttga attgggtttg gagtttccca atcttcctta ttatattgat ggtgatgtta
     3481 aattaacaca gtctatggcc atcatacgtt atatagctga caagcacaac atgttgggtg
     3541 gttgtccaaa agagcgtgca gagatttcaa tgcttgaagg agcggttttg gatattagat
     3601 acggtgtttc gagaattgca tatagtaaag actttgaaac tctcaaagtt gattttctta
     3661 gcaagctacc tgaaatgctg aaaatgttcg aagatcgttt atgtcataaa acatatttaa
     3721 atggtgatca tgtaacccat cctgacttca tgttgtatga cgctcttgat gttgttttat
     3781 acatggaccc aatgtgcctg gatgcgttcc caaaattagt ttgttttaaa aaacgtattg
     3841 aagctatccc acaaattgat aagtacttga aatccagcaa gtatatagca tggcctttgc
     3901 agggctggca agccacgttt ggtggtggcg accatcctcc aaaatcgctc gaggaaaacc
     3961 tgtattttca gggcgcgtgg tcccatcctc agttcgagaa aggcgcgggt tcaggtgctg
     4021 gcagtggagc gtccgcgtgg agtcatccgc aattcgaaaa atgagatccg gctgctaaca
     4081 aagcccgaaa ggaagctgag ttggctgctg ccaccgctga gcaataacta gcataacccc
     4141 ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa aggaggaact atatccggat
     4201 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     4261 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     4321 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     4381 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     4441 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     4501 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     4561 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     4621 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt
     4681 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     4741 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     4801 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     4861 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     4921 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     4981 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     5041 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     5101 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     5161 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     5221 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     5281 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     5341 gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     5401 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     5461 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     5521 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     5581 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     5641 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     5701 aaacttcatt tttaatttaa aaggatctag gtgaagatc
//
