LOCUS       sequence-344772-        6696 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  express E. coli E69 (O9a:K30) Wzc_K540M with 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: pBN082
FEATURES             Location/Qualifiers
     source          1..6696
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      59..86
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        105..196
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(197..265,266..1057)
                     /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(415..434)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1099..1554
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1186..1205)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1396..1417
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      1665..2253
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2154..2173
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2407..2424
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     2590..2612
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2750..2769
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(2949..3827)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        3854..4138
                     /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     4061..4080
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             6336..6353
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6407..6424)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      6577..6663
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 cggcccggag ggtggcgggc aggacgcccg ccataaactg ccaggcatca aattaagcag
       61 aaggccatcc tgacggatgg cctttttgcg tttctacaaa ctcttttgtt tatttttcta
      121 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
      181 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
      241 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
      301 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
      361 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
      421 tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
      481 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
      541 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
      601 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
      661 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
      721 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
      781 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
      841 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
      901 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
      961 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     1021 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     1081 tatactttag attgatttac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     1141 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     1201 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     1261 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg
     1321 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     1381 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     1441 gctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     1501 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca atttaaaagg
     1561 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     1621 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     1681 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     1741 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     1801 ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     1861 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     1921 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     1981 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     2041 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     2101 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     2161 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     2221 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     2281 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     2341 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     2401 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt
     2461 acgcatctgt gcggtatttc acaccgcata gggtcatggc tgcgccccga cacccgccaa
     2521 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     2581 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     2641 ggcagaagga gatggcgccc aacagtcccc cggccacggg gcctgccacc atacccacgc
     2701 cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg
     2761 cgatataggc gccagcaacc gcacctgtgg cgccggtgat gccggccacg atgcgtccgg
     2821 cgtagaggat ctgctcatgt ttgacagctt atcatcgatg cataatgtgc ctgtcaaatg
     2881 gacgaagcag ggattctgca aaccctatgc tactccgtca agccgtcaat tgtctgattc
     2941 gttaccaatt atgacaactt gacggctaca tcattcactt tttcttcaca accggcacgg
     3001 aactcgctcg ggctggcccc ggtgcatttt ttaaataccc gcgagaaata gagttgatcg
     3061 tcaaaaccaa cattgcgacc gacggtggcg ataggcatcc gggtggtgct caaaagcagc
     3121 ttcgcctggc tgatacgttg gtcctcgcgc cagcttaaga cgctaatccc taactgctgg
     3181 cggaaaagat gtgacagacg cgacggcgac aagcaaacat gctgtgcgac gctggcgata
     3241 tcaaaattgc tgtctgccag gtgatcgctg atgtactgac aagcctcgcg tacccgatta
     3301 tccatcggtg gatggagcga ctcgttaatc gcttccatgc gccgcagtaa caattgctca
     3361 agcagattta tcgccagcag ctccgaatag cgcccttccc cttgcccggc gttaatgatt
     3421 tgcccaaaca ggtcgctgaa atgcggctgg tgcgcttcat ccgggcgaaa gaaccccgta
     3481 ttggcaaata ttgacggcca gttaagccat tcatgccagt aggcgcgcgg acgaaagtaa
     3541 acccactggt gataccattc gcgagcctcc ggatgacgac cgtagtgatg aatctctcct
     3601 ggcgggaaca gcaaaatatc acccggtcgg caaacaaatt ctcgtccctg atttttcacc
     3661 accccctgac cgcgaatggt gagattgaga atataacctt tcattcccag cggtcggtcg
     3721 ataaaaaaat cgagataacc gttggcctca atcggcgtta aacccgccac cagatgggca
     3781 ttaaacgagt atcccggcag caggggatca ttttgcgctt cagccatact tttcatactc
     3841 ccgccattca gagaagaaac caattgtcca tattgcatca gacattgccg tcactgcgtc
     3901 ttttactggc tcttctcgct aaccaaaccg gtaaccccgc ttattaaaag cattctgtaa
     3961 caaagcggga ccaaagccat gacaaaaacg cgtaacaaaa gtgtctataa tcacggcaga
     4021 aaagtccaca ttgattattt gcacggcgtc acactttgct atgccatagc atttttatcc
     4081 ataagattag cggatcctac ctgacgcttt ttatcgcaac tctctactgt ttctccatac
     4141 ccgttttttt gggctagcag gaggaattca ccatgacttc agtcacatct aagcaatcaa
     4201 cgatactagg atctgatgaa atagatttag gtcgcgttat cggtgagcta attgatcatc
     4261 gtaagcttat tatatctata acatcggttt ttaccttatt tgcaatactc tatgcattat
     4321 tagcgactcc tatctatgag acggatgcgc taattcaaat agagcagaag caaggtaatg
     4381 ctatactaag cagtctaagt caggttttac ctgatggcca accccaatct gcacctgaaa
     4441 cagctttgct acaatctcga atgattcttg gcaaaactat agatgattta aaccttcaga
     4501 ttcaaattga acaaaaatat tttcctgtta ttggccgtgg gttagcaaga ttaatgggag
     4561 aaaaaccagg taatattgat attactcggt tatatttgcc tgatagtgac gatattagta
     4621 ataatactcc tagtataata ttaacagtaa aagataaaga aaattattcg atcaatagcg
     4681 atggtattca actcaatggg gttgtaggga cgttattaaa tgaaaaaggg atatcattac
     4741 tggttaacga aatcgatgct aaacctggag atcaatttgt tattacacag ttgcctcgat
     4801 taaaggccat ttctgatctt ttaaaaagtt ttagcgttgc tgatttgggt aaagatacgg
     4861 gaatgttaac gttgactctt actggagata acccaaaaag aatatctcat attttggaca
     4921 gtatcagtca gaactatctt gcacagaaca ttgctagaca agctgcgcag gatgctaaaa
     4981 gtctcgagtt tttaaatcaa caattaccca aagttcgtgc cgaattagat tctgcggaag
     5041 acaagttaaa tgcttatcgt aagcaaaaag actctgttga cttgaatatg gaagctaaat
     5101 cagtacttga tcaaatagtt aacgttgata atcaactaaa tgaattaacc ttccgtgagg
     5161 cagaagtttc acagctatat actaaggaac atcctactta taaggcgtta atggaaaaac
     5221 ggcaaacact tcaagaagaa aaatctaaac ttaataagcg cgttagtagt atgccctcga
     5281 cccagcagga ggttctgcga ttaagtcgag atgttgaatc tggtcgtgct gtatatttgc
     5341 aattgctaaa tcggcaacaa gaattaaaca ttgccaaatc aagtgctatt ggcaacgtta
     5401 gaattattga taatgctgta acagatccta atcctgtaag accaaagaag actatcataa
     5461 ttgtaatagg tgttgtttta gggctaatcg tttcggttgt actggtattg ttccaagtgt
     5521 ttctacgccg tggtatcgaa tccccagagc aactcgaaga aatcggaata aacgtgtatg
     5581 caagtattcc aatatctgaa tggcttacaa aaaatgctcg tcagtcaggg aaagtcagaa
     5641 aaaatcagtc agacacattg cttgctgttg gtaatcctgc cgatttagct gttgaggcca
     5701 ttcgtggttt acgcacaagt ttgcattttg caatgatgga agcaaaaaat aatgtcctaa
     5761 tgatatcggg tgctagtcca agcgcaggta tgacttttat tagcagtaac ttggcagcca
     5821 ctatagcaat tacagggaaa aaagttttat tcattgatgc tgaccttcgt aagggatatg
     5881 ctcataaaat gtttggtcat aaaaacgata aaggattatc tgagttttta tctggacaag
     5941 ccgctgctga aatgatcatt gataaagtag aagggggggg atttgattac attggtcgtg
     6001 gacaaattcc tccaaaccca gctgaattac ttatgcatcc gcgttttgaa caattattaa
     6061 attgggcctc acaaaattat gatttaatta taatagatac acctccaatt ttggcagtga
     6121 cagatgctgc gattattggt cgatatgctg gaacatgtct attagtcgct cgctttgaaa
     6181 aaaatactgt taaagaaatt gatgtaagta tgaagcggtt tgagcaaagt ggtgtggtag
     6241 tcaaaggctg tatacttaac ggtgttgtaa agaaagctag cagctacgaa agatatggcc
     6301 ataatcatta tggctattct tattatgata aaaaacacca ccaccaccac cactgactgc
     6361 aggcatgcaa gcttggctgt tttggcggat gagagaagat tttcagcctg atacagatta
     6421 aatcagaacg cagaagcggt ctgataaaac agaatttgcc tggcggcagt agcgcggtgg
     6481 tcccacctga ccccatgccg aactcagaag tgaaacgccg tagcgccgat ggtagtgtgg
     6541 ggtctcccca tgcgagagta gggaactgcc aggcatcaaa taaaacgaaa ggctcagtcg
     6601 aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcct gagtaggaca
     6661 aatccgccgg gagcggattt gaacgttgcg aagcaa
//
