LOCUS       sequence-344777-        6696 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  express E. coli E69 (O9a:K30) Wzc_K540M with Y705E, Y706E.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6696)
  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 6696)
  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: pBN083
FEATURES             Location/Qualifiers
     source          1..6696
                     /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             2483..2500
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2554..2571)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2724..2810
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2902..2929
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2948..3039
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3040..3108,3109..3900)
                     /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(3258..3277)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3942..4397
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4029..4048)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4239..4260
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      4508..5096
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4997..5016
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5250..5267
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     5433..5455
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5593..5612
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(5792..6670)
                     /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 gctaaaagtc tcgagttttt
     1141 aaatcaacaa ttacccaaag ttcgtgccga attagattct gcggaagaca agttaaatgc
     1201 ttatcgtaag caaaaagact ctgttgactt gaatatggaa gctaaatcag tacttgatca
     1261 aatagttaac gttgataatc aactaaatga attaaccttc cgtgaggcag aagtttcaca
     1321 gctatatact aaggaacatc ctacttataa ggcgttaatg gaaaaacggc aaacacttca
     1381 agaagaaaaa tctaaactta ataagcgcgt tagtagtatg ccctcgaccc agcaggaggt
     1441 tctgcgatta agtcgagatg ttgaatctgg tcgtgctgta tatttgcaat tgctaaatcg
     1501 gcaacaagaa ttaaacattg ccaaatcaag tgctattggc aacgttagaa ttattgataa
     1561 tgctgtaaca gatcctaatc ctgtaagacc aaagaagact atcataattg taataggtgt
     1621 tgttttaggg ctaatcgttt cggttgtact ggtattgttc caagtgtttc tacgccgtgg
     1681 tatcgaatcc ccagagcaac tcgaagaaat cggaataaac gtgtatgcaa gtattccaat
     1741 atctgaatgg cttacaaaaa atgctcgtca gtcagggaaa gtcagaaaaa atcagtcaga
     1801 cacattgctt gctgttggta atcctgccga tttagctgtt gaggccattc gtggtttacg
     1861 cacaagtttg cattttgcaa tgatggaagc aaaaaataat gtcctaatga tatcgggtgc
     1921 tagtccaagc gcaggtatga cttttattag cagtaacttg gcagccacta tagcaattac
     1981 agggaaaaaa gttttattca ttgatgctga ccttcgtaag ggatatgctc ataaaatgtt
     2041 tggtcataaa aacgataaag gattatctga gtttttatct ggacaagccg ctgctgaaat
     2101 gatcattgat aaagtagaag gggggggatt tgattacatt ggtcgtggac aaattcctcc
     2161 aaacccagct gaattactta tgcatccgcg ttttgaacaa ttattaaatt gggcctcaca
     2221 aaattatgat ttaattataa tagatacacc tccaattttg gcagtgacag atgctgcgat
     2281 tattggtcga tatgctggaa catgtctatt agtcgctcgc tttgaaaaaa atactgttaa
     2341 agaaattgat gtaagtatga agcggtttga gcaaagtggt gtggtagtca aaggctgtat
     2401 acttaacggt gttgtaaaga aagctagcag cgaggaaaga tatggccata atcattatgg
     2461 ctattcttat tatgataaaa aacaccacca ccaccaccac tgactgcagg catgcaagct
     2521 tggctgtttt ggcggatgag agaagatttt cagcctgata cagattaaat cagaacgcag
     2581 aagcggtctg ataaaacaga atttgcctgg cggcagtagc gcggtggtcc cacctgaccc
     2641 catgccgaac tcagaagtga aacgccgtag cgccgatggt agtgtggggt ctccccatgc
     2701 gagagtaggg aactgccagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct
     2761 ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag taggacaaat ccgccgggag
     2821 cggatttgaa cgttgcgaag caacggcccg gagggtggcg ggcaggacgc ccgccataaa
     2881 ctgccaggca tcaaattaag cagaaggcca tcctgacgga tggccttttt gcgtttctac
     2941 aaactctttt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     3001 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     3061 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     3121 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     3181 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     3241 agcactttta aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag
     3301 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     3361 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     3421 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     3481 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     3541 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     3601 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     3661 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     3721 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     3781 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     3841 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     3901 ctgtcagacc aagtttactc atatatactt tagattgatt tacgcgccct gtagcggcgc
     3961 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     4021 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     4081 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     4141 ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     4201 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     4261 aacaacactc aaccctatct cgggctattc ttttgattta taagggattt tgccgatttc
     4321 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     4381 attaacgttt acaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     4441 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     4501 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     4561 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     4621 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
     4681 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     4741 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     4801 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     4861 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     4921 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     4981 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     5041 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     5101 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     5161 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     5221 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     5281 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atagggtcat
     5341 ggctgcgccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     5401 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     5461 accgtcatca ccgaaacgcg cgaggcagaa ggagatggcg cccaacagtc ccccggccac
     5521 ggggcctgcc accataccca cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg
     5581 atcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg tggcgccggt
     5641 gatgccggcc acgatgcgtc cggcgtagag gatctgctca tgtttgacag cttatcatcg
     5701 atgcataatg tgcctgtcaa atggacgaag cagggattct gcaaacccta tgctactccg
     5761 tcaagccgtc aattgtctga ttcgttacca attatgacaa cttgacggct acatcattca
     5821 ctttttcttc acaaccggca cggaactcgc tcgggctggc cccggtgcat tttttaaata
     5881 cccgcgagaa atagagttga tcgtcaaaac caacattgcg accgacggtg gcgataggca
     5941 tccgggtggt gctcaaaagc agcttcgcct ggctgatacg ttggtcctcg cgccagctta
     6001 agacgctaat ccctaactgc tggcggaaaa gatgtgacag acgcgacggc gacaagcaaa
     6061 catgctgtgc gacgctggcg atatcaaaat tgctgtctgc caggtgatcg ctgatgtact
     6121 gacaagcctc gcgtacccga ttatccatcg gtggatggag cgactcgtta atcgcttcca
     6181 tgcgccgcag taacaattgc tcaagcagat ttatcgccag cagctccgaa tagcgccctt
     6241 ccccttgccc ggcgttaatg atttgcccaa acaggtcgct gaaatgcggc tggtgcgctt
     6301 catccgggcg aaagaacccc gtattggcaa atattgacgg ccagttaagc cattcatgcc
     6361 agtaggcgcg cggacgaaag taaacccact ggtgatacca ttcgcgagcc tccggatgac
     6421 gaccgtagtg atgaatctct cctggcggga acagcaaaat atcacccggt cggcaaacaa
     6481 attctcgtcc ctgatttttc accaccccct gaccgcgaat ggtgagattg agaatataac
     6541 ctttcattcc cagcggtcgg tcgataaaaa aatcgagata accgttggcc tcaatcggcg
     6601 ttaaacccgc caccagatgg gcattaaacg agtatcccgg cagcagggga tcattttgcg
     6661 cttcagccat acttttcata ctcccgccat tcagag
//
