LOCUS       sequence-344771-        6696 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  express E. coli E69 (O9a:K30) Wzc_K540M with Y713E, Y715E, Y717E, 
            Y718E mutations.
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: pBN202
FEATURES             Location/Qualifiers
     source          1..6696
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(57..143)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     296..313
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     CDS             complement(367..384)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        complement(2582..2866)
                     /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     complement(2640..2659)
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             2893..3771
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     primer_bind     complement(3951..3970)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(4108..4130)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4296..4313)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4467..5055)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4547..4566)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     rep_origin      complement(5166..5621)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5303..5324)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5515..5534
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(join(5663..6454,6455..6523))
                     /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     6286..6305
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6524..6615)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      complement(6634..6661)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 cctgcccgcc accctccggg ccgttgcttc gcaacgttca aatccgctcc cggcggattt
       61 gtcctactca ggagagcgtt caccgacaaa caacagataa aacgaaaggc ccagtctttc
      121 gactgagcct ttcgttttat ttgatgcctg gcagttccct actctcgcat ggggagaccc
      181 cacactacca tcggcgctac ggcgtttcac ttctgagttc ggcatggggt caggtgggac
      241 caccgcgcta ctgccgccag gcaaattctg ttttatcaga ccgcttctgc gttctgattt
      301 aatctgtatc aggctgaaaa tcttctctca tccgccaaaa cagccaagct tgcatgcctg
      361 cagtcagtgg tggtggtggt ggtgtttttt atcttcctca gactcgcctt catgattatg
      421 gccatatcta tagtagctgc tagctttctt tacaacaccg ttaagtatac agcctttgac
      481 taccacacca ctttgctcaa accgcttcat acttacatca atttctttaa cagtattttt
      541 ttcaaagcga gcgactaata gacatgttcc agcatatcga ccaataatcg cagcatctgt
      601 cactgccaaa attggaggtg tatctattat aattaaatca taattttgtg aggcccaatt
      661 taataattgt tcaaaacgcg gatgcataag taattcagct gggtttggag gaatttgtcc
      721 acgaccaatg taatcaaatc cccccccttc tactttatca atgatcattt cagcagcggc
      781 ttgtccagat aaaaactcag ataatccttt atcgttttta tgaccaaaca ttttatgagc
      841 atatccctta cgaaggtcag catcaatgaa taaaactttt ttccctgtaa ttgctatagt
      901 ggctgccaag ttactgctaa taaaagtcat acctgcgctt ggactagcac ccgatatcat
      961 taggacatta ttttttgctt ccatcattgc aaaatgcaaa cttgtgcgta aaccacgaat
     1021 ggcctcaaca gctaaatcgg caggattacc aacagcaagc aatgtgtctg actgattttt
     1081 tctgactttc cctgactgac gagcattttt tgtaagccat tcagatattg gaatacttgc
     1141 atacacgttt attccgattt cttcgagttg ctctggggat tcgataccac ggcgtagaaa
     1201 cacttggaac aataccagta caaccgaaac gattagccct aaaacaacac ctattacaat
     1261 tatgatagtc ttctttggtc ttacaggatt aggatctgtt acagcattat caataattct
     1321 aacgttgcca atagcacttg atttggcaat gtttaattct tgttgccgat ttagcaattg
     1381 caaatataca gcacgaccag attcaacatc tcgacttaat cgcagaacct cctgctgggt
     1441 cgagggcata ctactaacgc gcttattaag tttagatttt tcttcttgaa gtgtttgccg
     1501 tttttccatt aacgccttat aagtaggatg ttccttagta tatagctgtg aaacttctgc
     1561 ctcacggaag gttaattcat ttagttgatt atcaacgtta actatttgat caagtactga
     1621 tttagcttcc atattcaagt caacagagtc tttttgctta cgataagcat ttaacttgtc
     1681 ttccgcagaa tctaattcgg cacgaacttt gggtaattgt tgatttaaaa actcgagact
     1741 tttagcatcc tgcgcagctt gtctagcaat gttctgtgca agatagttct gactgatact
     1801 gtccaaaata tgagatattc tttttgggtt atctccagta agagtcaacg ttaacattcc
     1861 cgtatcttta cccaaatcag caacgctaaa actttttaaa agatcagaaa tggcctttaa
     1921 tcgaggcaac tgtgtaataa caaattgatc tccaggttta gcatcgattt cgttaaccag
     1981 taatgatatc cctttttcat ttaataacgt ccctacaacc ccattgagtt gaataccatc
     2041 gctattgatc gaataatttt ctttatcttt tactgttaat attatactag gagtattatt
     2101 actaatatcg tcactatcag gcaaatataa ccgagtaata tcaatattac ctggtttttc
     2161 tcccattaat cttgctaacc cacggccaat aacaggaaaa tatttttgtt caatttgaat
     2221 ctgaaggttt aaatcatcta tagttttgcc aagaatcatt cgagattgta gcaaagctgt
     2281 ttcaggtgca gattggggtt ggccatcagg taaaacctga cttagactgc ttagtatagc
     2341 attaccttgc ttctgctcta tttgaattag cgcatccgtc tcatagatag gagtcgctaa
     2401 taatgcatag agtattgcaa ataaggtaaa aaccgatgtt atagatataa taagcttacg
     2461 atgatcaatt agctcaccga taacgcgacc taaatctatt tcatcagatc ctagtatcgt
     2521 tgattgctta gatgtgactg aagtcatggt gaattcctcc tgctagccca aaaaaacggg
     2581 tatggagaaa cagtagagag ttgcgataaa aagcgtcagg taggatccgc taatcttatg
     2641 gataaaaatg ctatggcata gcaaagtgtg acgccgtgca aataatcaat gtggactttt
     2701 ctgccgtgat tatagacact tttgttacgc gtttttgtca tggctttggt cccgctttgt
     2761 tacagaatgc ttttaataag cggggttacc ggtttggtta gcgagaagag ccagtaaaag
     2821 acgcagtgac ggcaatgtct gatgcaatat ggacaattgg tttcttctct gaatggcggg
     2881 agtatgaaaa gtatggctga agcgcaaaat gatcccctgc tgccgggata ctcgtttaat
     2941 gcccatctgg tggcgggttt aacgccgatt gaggccaacg gttatctcga tttttttatc
     3001 gaccgaccgc tgggaatgaa aggttatatt ctcaatctca ccattcgcgg tcagggggtg
     3061 gtgaaaaatc agggacgaga atttgtttgc cgaccgggtg atattttgct gttcccgcca
     3121 ggagagattc atcactacgg tcgtcatccg gaggctcgcg aatggtatca ccagtgggtt
     3181 tactttcgtc cgcgcgccta ctggcatgaa tggcttaact ggccgtcaat atttgccaat
     3241 acggggttct ttcgcccgga tgaagcgcac cagccgcatt tcagcgacct gtttgggcaa
     3301 atcattaacg ccgggcaagg ggaagggcgc tattcggagc tgctggcgat aaatctgctt
     3361 gagcaattgt tactgcggcg catggaagcg attaacgagt cgctccatcc accgatggat
     3421 aatcgggtac gcgaggcttg tcagtacatc agcgatcacc tggcagacag caattttgat
     3481 atcgccagcg tcgcacagca tgtttgcttg tcgccgtcgc gtctgtcaca tcttttccgc
     3541 cagcagttag ggattagcgt cttaagctgg cgcgaggacc aacgtatcag ccaggcgaag
     3601 ctgcttttga gcaccacccg gatgcctatc gccaccgtcg gtcgcaatgt tggttttgac
     3661 gatcaactct atttctcgcg ggtatttaaa aaatgcaccg gggccagccc gagcgagttc
     3721 cgtgccggtt gtgaagaaaa agtgaatgat gtagccgtca agttgtcata attggtaacg
     3781 aatcagacaa ttgacggctt gacggagtag catagggttt gcagaatccc tgcttcgtcc
     3841 atttgacagg cacattatgc atcgatgata agctgtcaaa catgagcaga tcctctacgc
     3901 cggacgcatc gtggccggca tcaccggcgc cacaggtgcg gttgctggcg cctatatcgc
     3961 cgacatcacc gatggggaag atcgggctcg ccacttcggg ctcatgagcg cttgtttcgg
     4021 cgtgggtatg gtggcaggcc ccgtggccgg gggactgttg ggcgccatct ccttctgcct
     4081 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
     4141 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
     4201 tggcgggtgt cggggcgcag ccatgaccct atgcggtgtg aaataccgca cagatgcgta
     4261 aggagaaaat accgcatcag gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     4321 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     4381 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     4441 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     4501 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     4561 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     4621 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     4681 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     4741 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     4801 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     4861 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
     4921 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     4981 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     5041 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     5101 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     5161 cttttaaatt gtaaacgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     5221 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagcc
     5281 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     5341 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     5401 acccaaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     5461 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     5521 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     5581 caccacaccc gccgcgctta atgcgccgct acagggcgcg taaatcaatc taaagtatat
     5641 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     5701 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     5761 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     5821 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     5881 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     5941 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     6001 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     6061 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     6121 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     6181 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     6241 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaac acgggataat accgcgccac
     6301 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     6361 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     6421 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     6481 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     6541 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     6601 agaaaaataa acaaaagagt ttgtagaaac gcaaaaaggc catccgtcag gatggccttc
     6661 tgcttaattt gatgcctggc agtttatggc gggcgt
//
