LOCUS       sequence-452266-        8922 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of wzxE and other genes from the E. coli enterobacterial 
            common antigen biosynthesis gene cluster.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8922)
  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 8922)
  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: pTT021
FEATURES             Location/Qualifiers
     source          1..8922
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(96..974)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1001..1285
                     /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     1208..1227
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     primer_bind     complement(5780..5797)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      5950..6036
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      6128..6155
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        6174..6265
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6266..6334,6335..7126)
                     /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(6484..6503)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7168..7623
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7255..7274)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7465..7486
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      7734..8322
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8223..8242
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8476..8493
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     8659..8681
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     8819..8838
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 atcgatgcat aatgtgcctg tcaaatggac gaagcaggga ttctgcaaac cctatgctac
       61 tccgtcaagc cgtcaattgt ctgattcgtt accaattatg acaacttgac ggctacatca
      121 ttcacttttt cttcacaacc ggcacggaac tcgctcgggc tggccccggt gcatttttta
      181 aatacccgcg agaaatagag ttgatcgtca aaaccaacat tgcgaccgac ggtggcgata
      241 ggcatccggg tggtgctcaa aagcagcttc gcctggctga tacgttggtc ctcgcgccag
      301 cttaagacgc taatccctaa ctgctggcgg aaaagatgtg acagacgcga cggcgacaag
      361 caaacatgct gtgcgacgct ggcgatatca aaattgctgt ctgccaggtg atcgctgatg
      421 tactgacaag cctcgcgtac ccgattatcc atcggtggat ggagcgactc gttaatcgct
      481 tccatgcgcc gcagtaacaa ttgctcaagc agatttatcg ccagcagctc cgaatagcgc
      541 ccttcccctt gcccggcgtt aatgatttgc ccaaacaggt cgctgaaatg cggctggtgc
      601 gcttcatccg ggcgaaagaa ccccgtattg gcaaatattg acggccagtt aagccattca
      661 tgccagtagg cgcgcggacg aaagtaaacc cactggtgat accattcgcg agcctccgga
      721 tgacgaccgt agtgatgaat ctctcctggc gggaacagca aaatatcacc cggtcggcaa
      781 acaaattctc gtccctgatt tttcaccacc ccctgaccgc gaatggtgag attgagaata
      841 taacctttca ttcccagcgg tcggtcgata aaaaaatcga gataaccgtt ggcctcaatc
      901 ggcgttaaac ccgccaccag atgggcatta aacgagtatc ccggcagcag gggatcattt
      961 tgcgcttcag ccatactttt catactcccg ccattcagag aagaaaccaa ttgtccatat
     1021 tgcatcagac attgccgtca ctgcgtcttt tactggctct tctcgctaac caaaccggta
     1081 accccgctta ttaaaagcat tctgtaacaa agcgggacca aagccatgac aaaaacgcgt
     1141 aacaaaagtg tctataatca cggcagaaaa gtccacattg attatttgca cggcgtcaca
     1201 ctttgctatg ccatagcatt tttatccata agattagcgg atcctacctg acgcttttta
     1261 tcgcaactct ctactgtttc tccatacccg tttttttggg ctagcaggag gaattcacca
     1321 tggtgtcgtt ggcaaaagcg tccttgtgga cggcggccag tacactggtc aagattggtg
     1381 ccgggttact ggtcggtaag ttgctggcgg tgtcatttgg tccggcgggg cttgggctgg
     1441 cggcaaattt ccgccagttg attaccgtgc tcggcgtgct tgccggggct ggcatcttta
     1501 acggtgtaac caaatacgtt gcccagtacc atgataatcc gcaacagctg cgccgcgtgg
     1561 tcggcacttc atcagcgatg gtacttggtt tctctacgct gatggcgctg gtttttgtgc
     1621 tggcagctgc gccaatcagc cagggattgt ttggtaatac cgactatcag gggctggtgc
     1681 gtttagtggc gctggtgcaa atggggatcg cctggggcaa cctgttactg gcgctgatga
     1741 aaggctttcg cgatgccgca ggtaatgcgt tatcgctgat tgtcggcagc ttgattggcg
     1801 ttctcgcgta ctacgtcagt taccgtttgg gcggttatga aggggcgttg ctgggtctgg
     1861 cgctgattcc cgcgctggtg gtaattcctg ccgccatcat gttgatcaaa cgtggtgtca
     1921 tcccgttaag ctatctgaaa cccagctggg ataacggtct ggcagggcag ttgagcaaat
     1981 ttacgctcat ggcgttgatt acgtcggtga ccttgcctgt tgcttacatc atgatgcgta
     2041 aactgctggc ggcgcagtat agctgggatg aggtggggat ctggcaaggg gtgagcagta
     2101 tttccgatgc ctacctgcaa tttattacgg catcgttcag cgtatatttg ctgcccacgt
     2161 tgtcgcggct aacggaaaag cgcgatatca cccgggaagt ggttaaatcg ctgaaattcg
     2221 tcttaccggc agtggcggcg gcgagtttta ccgtctggct gctgcgtgat tttgctatct
     2281 ggctgctgtt gtcgaataaa tttaccgcta tgcgcgatct ctttgcctgg cagttagtgg
     2341 gtgatgtgtt aaaagtgggc gcttatgtct ttggttatct ggtgatcgcc aaagcgtcac
     2401 tgcggtttta tattctggcg gaagtcagcc agttcacttt attgatggta tttgcccact
     2461 ggctaatccc tgcgcatggt gcactgggcg cggcgcaggc atatatggca acttatatcg
     2521 tctatttttc tctttgttgt ggcgtgtttt tactctggcg taggcgggca tgactgtact
     2581 gattcacgta ctgggatcgg atatccctca ccataaccga accgttttgc ggtttttcaa
     2641 tgacgcgctg gccgcgacga gcgagcacgc gcgcgagttt atggttgttg gcaaggacga
     2701 cggcttaagt gatagctgtc cggcgctttc tgtgcaattt ttccctggga aaaaatcgct
     2761 ggcggaagcg gtcatcgcga aagcaaaagc taaccgtcag cagcgttttt tcttccacgg
     2821 tcagttcaat cccacactgt ggctggctct gctgagtggt ggcattaagc ccagccagtt
     2881 tttctggcat atctgggggg cagacctgta tgagctttcc agtggcttga gatataagct
     2941 tttttaccca ctacgtcgcc tggcgcaaaa gcgagtcggc tgtgtatttg ccacccgcgg
     3001 cgatttgagc ttttttgcca aaacgcaccc aaaggtgcgg ggcgaactgc tgttcttccc
     3061 gacgcggatg gacccttcgc tcaatacgat ggcgaacgat cggcaacgtg aagggaaaat
     3121 gaccattctg gtggggaact ccggcgaccg cagcaatgag catattgctg ccttgcgcgc
     3181 cgttcatcag caatttggcg atacggtaaa agtggtggtg ccgatgggat atccgcctaa
     3241 taacgaagcg tacattgagg aagttcgtca ggcggggctg gagttattca gcgaagaaaa
     3301 tctacaaatt ctgagcgaaa aactggaatt tgacgcctat ctggcgctac ttcgtcagtg
     3361 cgatcttggt tactttattt ttgcccgcca gcagggcatt ggtacgctgt gcttactgat
     3421 tcaggcgggc attccttgtg tgcttaaccg ggaaaatccg ttctggcagg atatgacgga
     3481 acaacatttg ccggtgctgt ttactaccga cgatctcaac gaggatattg tgcgtgaagc
     3541 gcagcgccag ttggcgtcgg tggataaaaa caccattgcc ttctttagcc ctaactatct
     3601 acaaggctgg cagcgggcgt tggcgattgc cgccagggag gtcgcatgag tctgctgcaa
     3661 ttcagtggcc tgtttgttgt ctggctgctc tgcacgctgt ttattgccac gctgacctgg
     3721 tttgagtttc gccgtgtgcg ctttaacttc aatgtcttct tttcattgct gtttttgctc
     3781 acctttttct tcggcttccc gctgaccagc gtgctggtat ttcgctttga tgttggtgtc
     3841 gcgccgccag aaatcttgtt gcaggcgttg ctttctgcgg gctgcttcta cgcggtttac
     3901 tatgtcacct acaaaacccg cctacgcaaa cgcgttgctg atgtaccgcg ccgtccgctg
     3961 tttaccatga accgcgtgga gaccaatctt acgtgggtga tcctgatggg tatcgcgctg
     4021 gtaagcgtcg gcatcttctt catgcacaac ggctttttgc tgttccggct taactcctac
     4081 agtcagatct tttccagtga agtctccggc gtggcgttaa aacgcttctt ttactttttc
     4141 atcccggcga tgctggtggt ctactttctg cgccaggaca gcaaagcgtg gctgtttttc
     4201 ctcgtcagca cggtcgcctt tggcttgctg acttatatga ttgtcggcgg cactcgcgcc
     4261 aatatcatca tcgcattcgc tatcttcctg tttattggca ttattcgcgg ctggatttcg
     4321 ttgtggatgc tggcggcggc gggcgtgctg gggattgttg gcatgttctg gctggcacta
     4381 aaacgctatg gaatgaatgt gagcggcgat gaagcgttct atacgtttct ctatctcact
     4441 cgcgacacct tctcgccgtg ggagaatctg gcgttgctgt tgcagaacta cgacaacatc
     4501 gacttccagg gcctggctcc aattgtccgc gatttctatg tctttatccc ttcctggctg
     4561 tggccgggtc gcccgagtat ggtgctgaac tcagccaact actttacctg ggaagtgctg
     4621 aataaccact ccggactggc gatctcgcct acgcttatag gctcactggt ggtgatgggc
     4681 ggcgcgttgt tcatcccgct cggggcgatc gtggttggtc tgatcatcaa atggttcgac
     4741 tggctgtatg agctgggcaa ccgcgagcct aatcgctata aagctgcgat attgcacagt
     4801 ttctgctttg gggcgatctt caatatgatc gtgctggcgc gtgaagggct ggattcgttt
     4861 gtctcacgcg tggtcttttt tatcgtggtc ttcggcgcat gtctgatgat cgcaaaactg
     4921 ttgtactggc tttttgaaag cgccggactc attcataaac gtacaaaatc atcgctccgg
     4981 acgcaggttg aaggataaca atgaataaca acaccacggc accaacctat acgctgcgtg
     5041 gcttacagtt gattggttgg cgtgatatgc agcacgccct cgattatctg tttgctgacg
     5101 ggcagcttaa gcagggaacg ctggttgcca ttaatgctga aaaaatgctg actattgaag
     5161 ataacgccga ggtcagggag ttaattaacg ctgccgaatt taaatatgcg gatggcatca
     5221 gcgttgtacg ttcagtacgt aaaaagtacc cgcaggcgca ggtttcccgc gttgccggtg
     5281 ccgatctctg ggaagagctg atggcgcgcg caggcaaaga agggacgccg gtatttcttg
     5341 tgggcggtaa acctgaagtg ctggcgcaaa ctgaagctaa actgcgcaac cagtggaatg
     5401 tgaatatcgt tggcagtcag gatggttatt ttaaacccga gcagcgtcag gcgctgtttg
     5461 aacgcattca tgccagcggt gcgcaaatcg tcaccgttgc gatgggatcg ccaaagcagg
     5521 agatcatcat gcgcgactgc cgtctggtac atccagatgc gctgtatatg ggcgttggcg
     5581 ggacttacga tgttttcacc ggtcacgtaa aacgcgcacc gaaaatctgg caaacgctgg
     5641 ggctggagtg gctctaccgc ctgctttcgc agccgagccg cattaagcgt cagcttcgtt
     5701 tgctgcgtta tttacgctgg cactacaccg gcaacctatg aaagcttggc tgttttggcg
     5761 gatgagagaa gattttcagc ctgatacaga ttaaatcaga acgcagaagc ggtctgataa
     5821 aacagaattt gcctggcggc agtagcgcgg tggtcccacc tgaccccatg ccgaactcag
     5881 aagtgaaacg ccgtagcgcc gatggtagtg tggggtctcc ccatgcgaga gtagggaact
     5941 gccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt
     6001 tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc cgggagcgga tttgaacgtt
     6061 gcgaagcaac ggcccggagg gtggcgggca ggacgcccgc cataaactgc caggcatcaa
     6121 attaagcaga aggccatcct gacggatggc ctttttgcgt ttctacaaac tcttttgttt
     6181 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     6241 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     6301 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     6361 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     6421 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     6481 tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg caagagcaac tcggtcgccg
     6541 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     6601 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     6661 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     6721 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     6781 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     6841 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     6901 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     6961 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     7021 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     7081 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     7141 ttactcatat atactttaga ttgatttacg cgccctgtag cggcgcatta agcgcggcgg
     7201 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     7261 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     7321 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     7381 atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     7441 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     7501 ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     7561 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa
     7621 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     7681 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     7741 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     7801 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     7861 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
     7921 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     7981 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     8041 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     8101 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     8161 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     8221 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     8281 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     8341 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     8401 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     8461 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat
     8521 tttctcctta cgcatctgtg cggtatttca caccgcatag ggtcatggct gcgccccgac
     8581 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
     8641 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
     8701 aacgcgcgag gcagaaggag atggcgccca acagtccccc ggccacgggg cctgccacca
     8761 tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct tccccatcgg
     8821 tgatgtcggc gatataggcg ccagcaaccg cacctgtggc gccggtgatg ccggccacga
     8881 tgcgtccggc gtagaggatc tgctcatgtt tgacagctta tc
//
