LOCUS       sequence-344776-        6306 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  express E. coli E69 (O9a:K30) Wzc_K540M with periplasmic motif 3 
            truncation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6306)
  AUTHORS   Yang Y, Liu J, Clarke BR, Seidel L, Bolla JR, Ward PN, Zhang P, 
            Robinson CV, Whitfield C, Naismith JH
  TITLE     The molecular basis of regulation of bacterial capsule assembly by 
            Wzc.
  JOURNAL   Nat Commun. 2021 Jul 16;12(1):4349. doi: 10.1038/s41467-021-24652-1.
  PUBMED    34272394
REFERENCE   2  (bases 1 to 6306)
  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: "Nat 
            Commun."; date: "2021-07-16"; volume: "12(1):4349. doi"; pages: " 
            10.1038/s41467-021-24652-1"
COMMENT     Sequence Label: pBN093
FEATURES             Location/Qualifiers
     source          1..6306
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..285
                     /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     208..227
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             2093..2110
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2164..2181)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2334..2420
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2512..2539
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2558..2649
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2650..2718,2719..3510)
                     /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(2868..2887)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3552..4007
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3639..3658)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3849..3870
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      4118..4706
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4607..4626
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4860..4877
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     5043..5065
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5203..5222
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(5402..6280)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 aagaaaccaa ttgtccatat tgcatcagac attgccgtca ctgcgtcttt tactggctct
       61 tctcgctaac caaaccggta accccgctta ttaaaagcat tctgtaacaa agcgggacca
      121 aagccatgac aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa gtccacattg
      181 attatttgca cggcgtcaca ctttgctatg ccatagcatt tttatccata agattagcgg
      241 atcctacctg acgcttttta tcgcaactct ctactgtttc tccatacccg tttttttggg
      301 ctagcaggag gaattcacca tgacttcagt cacatctaag caatcaacga tactaggatc
      361 tgatgaaata gatttaggtc gcgttatcgg tgagctaatt gatcatcgta agcttattat
      421 atctataaca tcggttttta ccttatttgc aatactctat gcattattag cgactcctat
      481 ctatgagacg gatgcgctaa ttcaaataga gcagaagcaa ggtaatgcta tactaagcag
      541 tctaagtcag gttttacctg atggccaacc ccaatctgca cctgaaacag ctttgctaca
      601 atctcgaatg attcttggca aaactataga tgatttaaac cttcagattc aaattgaaca
      661 aaaatatttt cctgttattg gccgtgggtt agcaagatta atgggagaaa aaccaggtaa
      721 tattgatatt actcggttat atttgcctga tagtgacgat attagtaata atactcctag
      781 tataatatta acagtaaaag ataaagaaaa ttattcgatc aatagcgatg gtattcaact
      841 caatggggtt gtagggacgt tattaaatga aaaagggata tcattactgg ttaacgaaat
      901 cgatgctaaa cctggagatc aatttgttat tacacagttg cctcgattaa aggccatttc
      961 tgatctttta aaaagtttta gcgttgctga tttgggtaaa gatacgggaa tgttaacgtt
     1021 gactcttact ggagataacc caaaaagaat atctcatatt ttggacagta tcagtcagaa
     1081 ctatcttgca cagaacattg ctagacaagc tgcgcaggat ggagcaggag ccaaatcaag
     1141 tgctattggc aacgttagaa ttattgataa tgctgtaaca gatcctaatc ctgtaagacc
     1201 aaagaagact atcataattg taataggtgt tgttttaggg ctaatcgttt cggttgtact
     1261 ggtattgttc caagtgtttc tacgccgtgg tatcgaatcc ccagagcaac tcgaagaaat
     1321 cggaataaac gtgtatgcaa gtattccaat atctgaatgg cttacaaaaa atgctcgtca
     1381 gtcagggaaa gtcagaaaaa atcagtcaga cacattgctt gctgttggta atcctgccga
     1441 tttagctgtt gaggccattc gtggtttacg cacaagtttg cattttgcaa tgatggaagc
     1501 aaaaaataat gtcctaatga tatcgggtgc tagtccaagc gcaggtatga cttttattag
     1561 cagtaacttg gcagccacta tagcaattac agggaaaaaa gttttattca ttgatgctga
     1621 ccttcgtaag ggatatgctc ataaaatgtt tggtcataaa aacgataaag gattatctga
     1681 gtttttatct ggacaagccg ctgctgaaat gatcattgat aaagtagaag gggggggatt
     1741 tgattacatt ggtcgtggac aaattcctcc aaacccagct gaattactta tgcatccgcg
     1801 ttttgaacaa ttattaaatt gggcctcaca aaattatgat ttaattataa tagatacacc
     1861 tccaattttg gcagtgacag atgctgcgat tattggtcga tatgctggaa catgtctatt
     1921 agtcgctcgc tttgaaaaaa atactgttaa agaaattgat gtaagtatga agcggtttga
     1981 gcaaagtggt gtggtagtca aaggctgtat acttaacggt gttgtaaaga aagctagcag
     2041 ctactataga tatggccata atcattatgg ctattcttat tatgataaaa aacaccacca
     2101 ccaccaccac tgactgcagg catgcaagct tggctgtttt ggcggatgag agaagatttt
     2161 cagcctgata cagattaaat cagaacgcag aagcggtctg ataaaacaga atttgcctgg
     2221 cggcagtagc gcggtggtcc cacctgaccc catgccgaac tcagaagtga aacgccgtag
     2281 cgccgatggt agtgtggggt ctccccatgc gagagtaggg aactgccagg catcaaataa
     2341 aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg
     2401 ctctcctgag taggacaaat ccgccgggag cggatttgaa cgttgcgaag caacggcccg
     2461 gagggtggcg ggcaggacgc ccgccataaa ctgccaggca tcaaattaag cagaaggcca
     2521 tcctgacgga tggccttttt gcgtttctac aaactctttt gtttattttt ctaaatacat
     2581 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     2641 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     2701 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     2761 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     2821 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     2881 gtattatccc gtgttgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     2941 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     3001 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     3061 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     3121 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     3181 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     3241 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     3301 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     3361 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     3421 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     3481 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     3541 tagattgatt tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     3601 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     3661 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     3721 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc
     3781 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     3841 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cgggctattc
     3901 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     3961 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaatttaaa aggatctagg
     4021 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     4081 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     4141 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     4201 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     4261 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     4321 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     4381 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     4441 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     4501 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     4561 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     4621 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     4681 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     4741 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     4801 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     4861 gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc
     4921 tgtgcggtat ttcacaccgc atagggtcat ggctgcgccc cgacacccgc caacacccgc
     4981 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     5041 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgaggcagaa
     5101 ggagatggcg cccaacagtc ccccggccac ggggcctgcc accataccca cgccgaaaca
     5161 agcgctcatg agcccgaagt ggcgagcccg atcttcccca tcggtgatgt cggcgatata
     5221 ggcgccagca accgcacctg tggcgccggt gatgccggcc acgatgcgtc cggcgtagag
     5281 gatctgctca tgtttgacag cttatcatcg atgcataatg tgcctgtcaa atggacgaag
     5341 cagggattct gcaaacccta tgctactccg tcaagccgtc aattgtctga ttcgttacca
     5401 attatgacaa cttgacggct acatcattca ctttttcttc acaaccggca cggaactcgc
     5461 tcgggctggc cccggtgcat tttttaaata cccgcgagaa atagagttga tcgtcaaaac
     5521 caacattgcg accgacggtg gcgataggca tccgggtggt gctcaaaagc agcttcgcct
     5581 ggctgatacg ttggtcctcg cgccagctta agacgctaat ccctaactgc tggcggaaaa
     5641 gatgtgacag acgcgacggc gacaagcaaa catgctgtgc gacgctggcg atatcaaaat
     5701 tgctgtctgc caggtgatcg ctgatgtact gacaagcctc gcgtacccga ttatccatcg
     5761 gtggatggag cgactcgtta atcgcttcca tgcgccgcag taacaattgc tcaagcagat
     5821 ttatcgccag cagctccgaa tagcgccctt ccccttgccc ggcgttaatg atttgcccaa
     5881 acaggtcgct gaaatgcggc tggtgcgctt catccgggcg aaagaacccc gtattggcaa
     5941 atattgacgg ccagttaagc cattcatgcc agtaggcgcg cggacgaaag taaacccact
     6001 ggtgatacca ttcgcgagcc tccggatgac gaccgtagtg atgaatctct cctggcggga
     6061 acagcaaaat atcacccggt cggcaaacaa attctcgtcc ctgatttttc accaccccct
     6121 gaccgcgaat ggtgagattg agaatataac ctttcattcc cagcggtcgg tcgataaaaa
     6181 aatcgagata accgttggcc tcaatcggcg ttaaacccgc caccagatgg gcattaaacg
     6241 agtatcccgg cagcagggga tcattttgcg cttcagccat acttttcata ctcccgccat
     6301 tcagag
//
