LOCUS       sequence-344774-        6696 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  express E. coli E69 (O9a:K30) Wzc_K540M with Y708E.
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: pBN084
FEATURES             Location/Qualifiers
     source          1..6696
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      33..60
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        79..170
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(171..239,240..1031)
                     /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(389..408)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1073..1528
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1160..1179)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1370..1391
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      1639..2227
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2128..2147
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2381..2398
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     2564..2586
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2724..2743
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(2923..3801)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        3828..4112
                     /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     4035..4054
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             6310..6327
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6381..6398)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      6551..6637
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 cccgccataa actgccaggc atcaaattaa gcagaaggcc atcctgacgg atggcctttt
       61 tgcgtttcta caaactcttt tgtttatttt tctaaataca ttcaaatatg tatccgctca
      121 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
      181 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
      241 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
      301 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
      361 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg
      421 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
      481 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
      541 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
      601 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
      661 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
      721 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
      781 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
      841 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
      901 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
      961 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     1021 agcattggta actgtcagac caagtttact catatatact ttagattgat ttacgcgccc
     1081 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     1141 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     1201 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     1261 cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc
     1321 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     1381 ttccaaactg gaacaacact caaccctatc tcgggctatt cttttgattt ataagggatt
     1441 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     1501 tttaacaaaa tattaacgtt tacaatttaa aaggatctag gtgaagatcc tttttgataa
     1561 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     1621 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     1681 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     1741 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
     1801 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     1861 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     1921 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     1981 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     2041 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     2101 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     2161 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     2221 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     2281 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     2341 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     2401 agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     2461 catagggtca tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     2521 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     2581 cagaggtttt caccgtcatc accgaaacgc gcgaggcaga aggagatggc gcccaacagt
     2641 cccccggcca cggggcctgc caccataccc acgccgaaac aagcgctcat gagcccgaag
     2701 tggcgagccc gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct
     2761 gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga ggatctgctc atgtttgaca
     2821 gcttatcatc gatgcataat gtgcctgtca aatggacgaa gcagggattc tgcaaaccct
     2881 atgctactcc gtcaagccgt caattgtctg attcgttacc aattatgaca acttgacggc
     2941 tacatcattc actttttctt cacaaccggc acggaactcg ctcgggctgg ccccggtgca
     3001 ttttttaaat acccgcgaga aatagagttg atcgtcaaaa ccaacattgc gaccgacggt
     3061 ggcgataggc atccgggtgg tgctcaaaag cagcttcgcc tggctgatac gttggtcctc
     3121 gcgccagctt aagacgctaa tccctaactg ctggcggaaa agatgtgaca gacgcgacgg
     3181 cgacaagcaa acatgctgtg cgacgctggc gatatcaaaa ttgctgtctg ccaggtgatc
     3241 gctgatgtac tgacaagcct cgcgtacccg attatccatc ggtggatgga gcgactcgtt
     3301 aatcgcttcc atgcgccgca gtaacaattg ctcaagcaga tttatcgcca gcagctccga
     3361 atagcgccct tccccttgcc cggcgttaat gatttgccca aacaggtcgc tgaaatgcgg
     3421 ctggtgcgct tcatccgggc gaaagaaccc cgtattggca aatattgacg gccagttaag
     3481 ccattcatgc cagtaggcgc gcggacgaaa gtaaacccac tggtgatacc attcgcgagc
     3541 ctccggatga cgaccgtagt gatgaatctc tcctggcggg aacagcaaaa tatcacccgg
     3601 tcggcaaaca aattctcgtc cctgattttt caccaccccc tgaccgcgaa tggtgagatt
     3661 gagaatataa cctttcattc ccagcggtcg gtcgataaaa aaatcgagat aaccgttggc
     3721 ctcaatcggc gttaaacccg ccaccagatg ggcattaaac gagtatcccg gcagcagggg
     3781 atcattttgc gcttcagcca tacttttcat actcccgcca ttcagagaag aaaccaattg
     3841 tccatattgc atcagacatt gccgtcactg cgtcttttac tggctcttct cgctaaccaa
     3901 accggtaacc ccgcttatta aaagcattct gtaacaaagc gggaccaaag ccatgacaaa
     3961 aacgcgtaac aaaagtgtct ataatcacgg cagaaaagtc cacattgatt atttgcacgg
     4021 cgtcacactt tgctatgcca tagcattttt atccataaga ttagcggatc ctacctgacg
     4081 ctttttatcg caactctcta ctgtttctcc atacccgttt ttttgggcta gcaggaggaa
     4141 ttcaccatga cttcagtcac atctaagcaa tcaacgatac taggatctga tgaaatagat
     4201 ttaggtcgcg ttatcggtga gctaattgat catcgtaagc ttattatatc tataacatcg
     4261 gtttttacct tatttgcaat actctatgca ttattagcga ctcctatcta tgagacggat
     4321 gcgctaattc aaatagagca gaagcaaggt aatgctatac taagcagtct aagtcaggtt
     4381 ttacctgatg gccaacccca atctgcacct gaaacagctt tgctacaatc tcgaatgatt
     4441 cttggcaaaa ctatagatga tttaaacctt cagattcaaa ttgaacaaaa atattttcct
     4501 gttattggcc gtgggttagc aagattaatg ggagaaaaac caggtaatat tgatattact
     4561 cggttatatt tgcctgatag tgacgatatt agtaataata ctcctagtat aatattaaca
     4621 gtaaaagata aagaaaatta ttcgatcaat agcgatggta ttcaactcaa tggggttgta
     4681 gggacgttat taaatgaaaa agggatatca ttactggtta acgaaatcga tgctaaacct
     4741 ggagatcaat ttgttattac acagttgcct cgattaaagg ccatttctga tcttttaaaa
     4801 agttttagcg ttgctgattt gggtaaagat acgggaatgt taacgttgac tcttactgga
     4861 gataacccaa aaagaatatc tcatattttg gacagtatca gtcagaacta tcttgcacag
     4921 aacattgcta gacaagctgc gcaggatgct aaaagtctcg agtttttaaa tcaacaatta
     4981 cccaaagttc gtgccgaatt agattctgcg gaagacaagt taaatgctta tcgtaagcaa
     5041 aaagactctg ttgacttgaa tatggaagct aaatcagtac ttgatcaaat agttaacgtt
     5101 gataatcaac taaatgaatt aaccttccgt gaggcagaag tttcacagct atatactaag
     5161 gaacatccta cttataaggc gttaatggaa aaacggcaaa cacttcaaga agaaaaatct
     5221 aaacttaata agcgcgttag tagtatgccc tcgacccagc aggaggttct gcgattaagt
     5281 cgagatgttg aatctggtcg tgctgtatat ttgcaattgc taaatcggca acaagaatta
     5341 aacattgcca aatcaagtgc tattggcaac gttagaatta ttgataatgc tgtaacagat
     5401 cctaatcctg taagaccaaa gaagactatc ataattgtaa taggtgttgt tttagggcta
     5461 atcgtttcgg ttgtactggt attgttccaa gtgtttctac gccgtggtat cgaatcccca
     5521 gagcaactcg aagaaatcgg aataaacgtg tatgcaagta ttccaatatc tgaatggctt
     5581 acaaaaaatg ctcgtcagtc agggaaagtc agaaaaaatc agtcagacac attgcttgct
     5641 gttggtaatc ctgccgattt agctgttgag gccattcgtg gtttacgcac aagtttgcat
     5701 tttgcaatga tggaagcaaa aaataatgtc ctaatgatat cgggtgctag tccaagcgca
     5761 ggtatgactt ttattagcag taacttggca gccactatag caattacagg gaaaaaagtt
     5821 ttattcattg atgctgacct tcgtaaggga tatgctcata aaatgtttgg tcataaaaac
     5881 gataaaggat tatctgagtt tttatctgga caagccgctg ctgaaatgat cattgataaa
     5941 gtagaagggg ggggatttga ttacattggt cgtggacaaa ttcctccaaa cccagctgaa
     6001 ttacttatgc atccgcgttt tgaacaatta ttaaattggg cctcacaaaa ttatgattta
     6061 attataatag atacacctcc aattttggca gtgacagatg ctgcgattat tggtcgatat
     6121 gctggaacat gtctattagt cgctcgcttt gaaaaaaata ctgttaaaga aattgatgta
     6181 agtatgaagc ggtttgagca aagtggtgtg gtagtcaaag gctgtatact taacggtgtt
     6241 gtaaagaaag ctagcagcta ctatagagaa ggccataatc attatggcta ttcttattat
     6301 gataaaaaac accaccacca ccaccactga ctgcaggcat gcaagcttgg ctgttttggc
     6361 ggatgagaga agattttcag cctgatacag attaaatcag aacgcagaag cggtctgata
     6421 aaacagaatt tgcctggcgg cagtagcgcg gtggtcccac ctgaccccat gccgaactca
     6481 gaagtgaaac gccgtagcgc cgatggtagt gtggggtctc cccatgcgag agtagggaac
     6541 tgccaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg
     6601 ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg ccgggagcgg atttgaacgt
     6661 tgcgaagcaa cggcccggag ggtggcgggc aggacg
//
