LOCUS       sequence-389767-        5954 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of C-terminally His10-tagged WzyE from Pectobacterium 
            atrosepticum in E.coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5954)
  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 5954)
  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-His
FEATURES             Location/Qualifiers
     source          1..5954
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..111
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(291..1169)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1196..1480
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     1403..1422
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             2910..2930
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2934..2963
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(3007..3024)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3177..3263
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3355..3382
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3401..3492
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3493..3561,3562..4353)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3711..3730)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4395..4850
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4482..4501)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4692..4713
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      4961..5549
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5450..5469
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5703..5720
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     5886..5908
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgctcatga
       61 gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa
      121 ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg atctgctcat
      181 gtttgacagc ttatcatcga tgcataatgt gcctgtcaaa tggacgaagc agggattctg
      241 caaaccctat gctactccgt caagccgtca attgtctgat tcgttaccaa ttatgacaac
      301 ttgacggcta catcattcac tttttcttca caaccggcac ggaactcgct cgggctggcc
      361 ccggtgcatt ttttaaatac ccgcgagaaa tagagttgat cgtcaaaacc aacattgcga
      421 ccgacggtgg cgataggcat ccgggtggtg ctcaaaagca gcttcgcctg gctgatacgt
      481 tggtcctcgc gccagcttaa gacgctaatc cctaactgct ggcggaaaag atgtgacaga
      541 cgcgacggcg acaagcaaac atgctgtgcg acgctggcga tatcaaaatt gctgtctgcc
      601 aggtgatcgc tgatgtactg acaagcctcg cgtacccgat tatccatcgg tggatggagc
      661 gactcgttaa tcgcttccat gcgccgcagt aacaattgct caagcagatt tatcgccagc
      721 agctccgaat agcgcccttc cccttgcccg gcgttaatga tttgcccaaa caggtcgctg
      781 aaatgcggct ggtgcgcttc atccgggcga aagaaccccg tattggcaaa tattgacggc
      841 cagttaagcc attcatgcca gtaggcgcgc ggacgaaagt aaacccactg gtgataccat
      901 tcgcgagcct ccggatgacg accgtagtga tgaatctctc ctggcgggaa cagcaaaata
      961 tcacccggtc ggcaaacaaa ttctcgtccc tgatttttca ccaccccctg accgcgaatg
     1021 gtgagattga gaatataacc tttcattccc agcggtcggt cgataaaaaa atcgagataa
     1081 ccgttggcct caatcggcgt taaacccgcc accagatggg cattaaacga gtatcccggc
     1141 agcaggggat cattttgcgc ttcagccata cttttcatac tcccgccatt cagagaagaa
     1201 accaattgtc catattgcat cagacattgc cgtcactgcg tcttttactg gctcttctcg
     1261 ctaaccaaac cggtaacccc gcttattaaa agcattctgt aacaaagcgg gaccaaagcc
     1321 atgacaaaaa cgcgtaacaa aagtgtctat aatcacggca gaaaagtcca cattgattat
     1381 ttgcacggcg tcacactttg ctatgccata gcatttttat ccataagatt agcggatctt
     1441 acctgacgct ttttatcgca actctctact gtttctccat acccgttttt ttgggctagc
     1501 aggaggaata caccatggcg cttgggcaat ttggagggct gtttgtcgtc tacctgattt
     1561 cagtcatctt catcctgagc ttgacctgga tggagtttcg gcgcgtacgt tttaacttta
     1621 acgtgctgtt ctcgctgctc tatttattga cgttctattt tggttttccc tttacctgcg
     1681 tgctggtgtt ccggtttggc gttgaagtcg tccccgtgca gtatctgctt caggcgatgt
     1741 tgtccgcaac ggcgttctat gctgtctatt acgtcagcta caagacgcga ctacgccaga
     1801 aaacctccgt gccgcgtgct ccactcctga cggtgaaccg agttgaagct aatctgacct
     1861 ggctgctgct ggcgttgatt gcggtagcca ccgtcggtat tttcttcctc aacaatggct
     1921 tcctgctgtt taagctgcgc tcttacagcc agatattctc tagcgatgtg tccggtgtgg
     1981 cgttgaaacg ctttttctac ttcttcattc cggcaatgct ggtggtgtat ttcctgcgcc
     2041 agacgcagcg agcgtggcta ctgttcctca tcggaacggt cgcatttgga atgctgacat
     2101 atgtgattgt cggcgggacg cgcgcgaatc tgatcattgc cttcgcccta tttctgttta
     2161 ttggcattgt gcgtggttgg attacgctct ggatgttggt cgctgctggg atatttggta
     2221 ttgtcgggat gttctggctg gcgctgaagc gctatgggct ggacgtcagc ggtgattacg
     2281 cattttatac cttcctctat ttaactcgtg acaccttctc gccctgggag aatctggcgc
     2341 tgctgtggca gaattatgac aagattgagt ttcaggggct ggcaccgatt gctcgtgatt
     2401 tctacgtctt tattccctcc tggctatggc ctgaccgtcc gaatctggtg ttaaacagcg
     2461 cgaactattt cacctgggaa gtgttgaata accactctgg gctggcgata tcccctacgc
     2521 tgttaggctc gctggtggtg atgggcggcg tgctatttat cccgttgggt gcgattgcgg
     2581 tagggctggt tatcaagtgg ttcgactggg tgtacgagtt agggaagaac gacagcaatc
     2641 gctataaggc cgccattttg caggcgttct gcttcggtgc ggtgttcaat attattgtgc
     2701 taacgcgcga aggcgtcgat tcctttgttt cacgtgtggt gtttttctgt ctgatatttg
     2761 gcctgtgcct gctcgttgcc aaactgcttt actggctgct ggaaagcgcc ggactcatcc
     2821 ggcagcgtct gaggcgcatg cgcgccacgc cgctcgcgcc gacacccaat accgtacagg
     2881 atccggtgat aaaggagcag atactcgagg aaaacctgta ttttcagggc gcgcaccacc
     2941 accaccacca tcaccatcac cattgataac gcttggctgt tttggcggat gagagaagat
     3001 tttcagcctg atacagatta aatcagaacg cagaagcggt ctgataaaac agaatttgcc
     3061 tggcggcagt agcgcggtgg tcccacctga ccccatgccg aactcagaag tgaaacgccg
     3121 tagcgccgat ggtagtgtgg ggtctcccca tgcgagagta gggaactgcc aggcatcaaa
     3181 taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga
     3241 acgctctcct gagtaggaca aatccgccgg gagcggattt gaacgttgcg aagcaacggc
     3301 ccggagggtg gcgggcagga cgcccgccat aaactgccag gcatcaaatt aagcagaagg
     3361 ccatcctgac ggatggcctt tttgcgtttc tacaaactct tttgtttatt tttctaaata
     3421 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     3481 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     3541 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     3601 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     3661 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     3721 gcggtattat cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     3781 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     3841 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     3901 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     3961 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     4021 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     4081 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     4141 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     4201 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     4261 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     4321 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     4381 ctttagattg atttacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     4441 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     4501 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     4561 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg
     4621 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     4681 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcgggcta
     4741 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     4801 ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt aaaaggatct
     4861 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     4921 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     4981 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     5041 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     5101 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     5161 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     5221 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     5281 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     5341 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     5401 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     5461 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     5521 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     5581 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     5641 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     5701 gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc
     5761 atctgtgcgg tatttcacac cgcatagggt catggctgcg ccccgacacc cgccaacacc
     5821 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     5881 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca
     5941 gaaggagatg gcgc
//
