LOCUS       sequence-452268-        5816 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of C-terminally TEV cleavable His10-tagged WzxE from E. 
            coli in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5816)
  AUTHORS   Le Bas A, Clarke BR, Teelucksingh T, Lee M, El Omari K, Giltrap AM, 
            McMahon SA, Liu H, Beale JH, Mykhaylyk V, Duman R, Paterson NG, Ward
            PN, Harrison PJ, Weckener M, Pardon E, Steyaert J, Liu H, Quigley A,
            Davis BG, Wagner A, Whitfield C, Naismith JH
  TITLE     Structure of WzxE the lipid III flippase for Enterobacterial Common 
            Antigen polysaccharide.
  JOURNAL   Open Biol. 2025 Jan;15(1):240310. doi: 10.1098/rsob.240310. Epub 
            2025 Jan 8.
  PUBMED    39772807
REFERENCE   2  (bases 1 to 5816)
  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: "2025-01"; volume: "15(1):240310. doi"; pages: " 
            10.1098/rsob.240310. Epub 2025 Jan 8"
COMMENT     Sequence Label: pBAD-WzxE-TEV-His10
FEATURES             Location/Qualifiers
     source          1..5816
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     54..73
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             1423..1443
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1447..1476
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(1520..1537)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1690..1776
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1868..1895
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1914..2005
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2006..2074,2075..2866)
                     /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(2224..2243)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2908..3363
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2995..3014)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3205..3226
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      3474..4062
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3963..3982
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4216..4233
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     4399..4421
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4559..4578
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(4758..5636)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        join(5663..5816,1..131)
                     /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)"
ORIGIN
        1 taatcacggc agaaaagtcc acattgatta tttgcacggc gtcacacttt gctatgccat
       61 agcattttta tccataagat tagcggatct tacctgacgc tttttatcgc aactctctac
      121 tgtttctcca tacccgtttt tttgggctag caggaggaat acaccatggt gtcgttggca
      181 aaagcgtcct tgtggacggc ggccagtaca ctggtcaaga ttggtgccgg gttactggtc
      241 ggtaagttgc tggcggtgtc atttggtccg gcggggcttg ggctggcggc aaatttccgc
      301 cagttgatta ccgtgctcgg cgtgcttgcc ggggctggca tctttaacgg tgtaaccaaa
      361 tacgttgccc agtaccatga taatccgcaa cagctgcgcc gcgtggtcgg cacttcatca
      421 gcgatggtac ttggtttctc tacgctgatg gcgctggttt ttgtgctggc agctgcgcca
      481 atcagccagg gattgtttgg taataccgac tatcaggggc tggtgcgttt agtggcgctg
      541 gtgcaaatgg ggatcgcctg gggcaacctg ttactggcgc tgatgaaagg ctttcgcgat
      601 gccgcaggta atgcgttatc gctgattgtc ggcagcttga ttggcgttct cgcgtactac
      661 gtcagttacc gtttgggcgg ttatgaaggg gcgttgctgg gtctggcgct gattcccgcg
      721 ctggtggtaa ttcctgccgc catcatgttg atcaaacgtg gtgtcatccc gttaagctat
      781 ctgaaaccca gctgggataa cggtctggca gggcagttga gcaaatttac gctcatggcg
      841 ttgattacgt cggtgacctt gcctgttgct tacatcatga tgcgtaaact gctggcggcg
      901 cagtatagct gggatgaggt ggggatctgg caaggggtga gcagtatttc cgatgcctac
      961 ctgcaattta ttacggcatc gttcagcgta tatttgctgc ccacgttgtc gcggctaacg
     1021 gaaaagcgcg atatcacccg ggaagtggtt aaatcgctga aattcgtctt accggcagtg
     1081 gcggcggcga gttttaccgt ctggctgctg cgtgattttg ctatctggct gctgttgtcg
     1141 aataaattta ccgctatgcg cgatctcttt gcctggcagt tagtgggtga tgtgttaaaa
     1201 gtgggcgctt atgtctttgg ttatctggtg atcgccaaag cgtcactgcg gttttatatt
     1261 ctggcggaag tcagccagtt cactttattg atggtatttg cccactggct aatccctgcg
     1321 catggtgcac tgggcgcggc gcaggcatat atggcaactt atatcgtcta tttttctctt
     1381 tgttgtggcg tgtttttact ctggcgtagg cgggcactcg aggaaaacct gtattttcag
     1441 ggcgcgcacc accaccacca ccatcaccat caccattgat aacgcttggc tgttttggcg
     1501 gatgagagaa gattttcagc ctgatacaga ttaaatcaga acgcagaagc ggtctgataa
     1561 aacagaattt gcctggcggc agtagcgcgg tggtcccacc tgaccccatg ccgaactcag
     1621 aagtgaaacg ccgtagcgcc gatggtagtg tggggtctcc ccatgcgaga gtagggaact
     1681 gccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt
     1741 tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc cgggagcgga tttgaacgtt
     1801 gcgaagcaac ggcccggagg gtggcgggca ggacgcccgc cataaactgc caggcatcaa
     1861 attaagcaga aggccatcct gacggatggc ctttttgcgt ttctacaaac tcttttgttt
     1921 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     1981 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     2041 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     2101 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     2161 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     2221 tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg caagagcaac tcggtcgccg
     2281 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     2341 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     2401 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     2461 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     2521 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     2581 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     2641 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     2701 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     2761 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     2821 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     2881 ttactcatat atactttaga ttgatttacg cgccctgtag cggcgcatta agcgcggcgg
     2941 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     3001 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     3061 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     3121 atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     3181 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     3241 ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     3301 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa
     3361 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     3421 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     3481 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     3541 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     3601 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
     3661 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     3721 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     3781 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     3841 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     3901 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     3961 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     4021 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     4081 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     4141 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     4201 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat
     4261 tttctcctta cgcatctgtg cggtatttca caccgcatag ggtcatggct gcgccccgac
     4321 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
     4381 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
     4441 aacgcgcgag gcagaaggag atggcgccca acagtccccc ggccacgggg cctgccacca
     4501 tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct tccccatcgg
     4561 tgatgtcggc gatataggcg ccagcaaccg cacctgtggc gccggtgatg ccggccacga
     4621 tgcgtccggc gtagaggatc tgctcatgtt tgacagctta tcatcgatgc ataatgtgcc
     4681 tgtcaaatgg acgaagcagg gattctgcaa accctatgct actccgtcaa gccgtcaatt
     4741 gtctgattcg ttaccaatta tgacaacttg acggctacat cattcacttt ttcttcacaa
     4801 ccggcacgga actcgctcgg gctggccccg gtgcattttt taaatacccg cgagaaatag
     4861 agttgatcgt caaaaccaac attgcgaccg acggtggcga taggcatccg ggtggtgctc
     4921 aaaagcagct tcgcctggct gatacgttgg tcctcgcgcc agcttaagac gctaatccct
     4981 aactgctggc ggaaaagatg tgacagacgc gacggcgaca agcaaacatg ctgtgcgacg
     5041 ctggcgatat caaaattgct gtctgccagg tgatcgctga tgtactgaca agcctcgcgt
     5101 acccgattat ccatcggtgg atggagcgac tcgttaatcg cttccatgcg ccgcagtaac
     5161 aattgctcaa gcagatttat cgccagcagc tccgaatagc gcccttcccc ttgcccggcg
     5221 ttaatgattt gcccaaacag gtcgctgaaa tgcggctggt gcgcttcatc cgggcgaaag
     5281 aaccccgtat tggcaaatat tgacggccag ttaagccatt catgccagta ggcgcgcgga
     5341 cgaaagtaaa cccactggtg ataccattcg cgagcctccg gatgacgacc gtagtgatga
     5401 atctctcctg gcgggaacag caaaatatca cccggtcggc aaacaaattc tcgtccctga
     5461 tttttcacca ccccctgacc gcgaatggtg agattgagaa tataaccttt cattcccagc
     5521 ggtcggtcga taaaaaaatc gagataaccg ttggcctcaa tcggcgttaa acccgccacc
     5581 agatgggcat taaacgagta tcccggcagc aggggatcat tttgcgcttc agccatactt
     5641 ttcatactcc cgccattcag agaagaaacc aattgtccat attgcatcag acattgccgt
     5701 cactgcgtct tttactggct cttctcgcta accaaaccgg taaccccgct tattaaaagc
     5761 attctgtaac aaagcgggac caaagccatg acaaaaacgc gtaacaaaag tgtcta
//
