LOCUS       sequence-398031-        5064 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of C-terminally distrep tag II-tagged WzyE from 
            Pectobacterium atrosepticum in E.coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5064)
  AUTHORS   Weckener M, Woodward LS, Clarke BR, Liu H, Ward PN, Le Bas A, Bhella
            D, Whitfield C, Naismith JH
  TITLE     The lipid linked oligosaccharide polymerase Wzy and its regulating 
            co-polymerase, Wzz, from enterobacterial common antigen biosynthesis
            form a complex.
  JOURNAL   Open Biol. 2023 Mar;13(3):220373. doi: 10.1098/rsob.220373. Epub 
            2023 Mar 22.
  PUBMED    36944376
REFERENCE   2  (bases 1 to 5064)
  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: "Open 
            Biol."; date: "2023-03"; volume: "13(3):220373. doi"; pages: " 
            10.1098/rsob.220373. Epub 2023 Mar 22"
COMMENT     Sequence Label: pWzyE-distrepII
FEATURES             Location/Qualifiers
     source          1..5064
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     239..256
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    271..410
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(336..355)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     496..518
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(512..703)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     1212..1231
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1212..1230
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1262..1284
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2686..2706
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2710..2733
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             2770..2793
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(2846..2864)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2860..2907
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2944..3399
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3031..3050)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3241..3262
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3425..3529
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3530..3598,3599..4390)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3748..3767)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(4561..5064,1..85)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     join(5050..5064,1..5)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
       61 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
      121 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
      181 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
      241 cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg tattttctcc ttacgcatct
      301 gtgcggtatt tcacaccgca atggtgcact ctcagtacaa tctgctctga tgccgcatag
      361 ttaagccagt atacactccg ctatcgctac gtgactgggt catggctgcg ccccgacacc
      421 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
      481 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
      541 gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg aagcgattca cagatgtctg
      601 cctgttcatc cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc tggcttctga
      661 taaagcgggc catgttaagg gcggtttttt cctgtttggt cactgatgcc tccgtgtaag
      721 ggggatttct gttcatgggg gtaatgatac cgatgaaacg agagaggatg ctcacgatac
      781 gggttactga tgatgaacat gcccggttac tggaacgttg tgagggtaaa caactggcgg
      841 tatggatgcg gcgggaccag agaaaaatca ctcagggtca atgccagcgc ttcgttaata
      901 cagatgtagg tgttccacag ggtagccagc agcatcctgc gatgcagatc cggaacataa
      961 tggtgcaggg cgctgacttc cgcgtttcca gactttacga aacacggaaa ccgaagacca
     1021 ttcatgttgt tgctcaggtc gcagacgttt tgcagcagca gtcgcttcac gttcgctcgc
     1081 gtatcggtga ttcattctgc taaccagtaa ggcaaccccg ccagcctagc cgggtcctca
     1141 acgacaggag cacgatcatg cgcacccgtg gccaggaccc aacgctgccc gagatctcga
     1201 tcccgcgaaa ttaatacgac tcactatagg gagaccacaa cggtttccct ctagaaataa
     1261 ttttgtttaa ctttaagaag gagatatacc atggcgcttg ggcaatttgg agggctgttt
     1321 gtcgtctacc tgatttcagt catcttcatc ctgagcttga cctggatgga gtttcggcgc
     1381 gtacgtttta actttaacgt gctgttctcg ctgctctatt tattgacgtt ctattttggt
     1441 tttcccttta cctgcgtgct ggtgttccgg tttggcgttg aagtcgtccc cgtgcagtat
     1501 ctgcttcagg cgatgttgtc cgcaacggcg ttctatgctg tctattacgt cagctacaag
     1561 acgcgactac gccagaaaac ctccgtgccg cgtgctccac tcctgacggt gaaccgagtt
     1621 gaagctaatc tgacctggct gctgctggcg ttgattgcgg tagccaccgt cggtattttc
     1681 ttcctcaaca atggcttcct gctgtttaag ctgcgctctt acagccagat attctctagc
     1741 gatgtgtccg gtgtggcgtt gaaacgcttt ttctacttct tcattccggc aatgctggtg
     1801 gtgtatttcc tgcgccagac gcagcgagcg tggctactgt tcctcatcgg aacggtcgca
     1861 tttggaatgc tgacatatgt gattgtcggc gggacgcgcg cgaatctgat cattgccttc
     1921 gccctatttc tgtttattgg cattgtgcgt ggttggatta cgctctggat gttggtcgct
     1981 gctgggatat ttggtattgt cgggatgttc tggctggcgc tgaagcgcta tgggctggac
     2041 gtcagcggtg attacgcatt ttataccttc ctctatttaa ctcgtgacac cttctcgccc
     2101 tgggagaatc tggcgctgct gtggcagaat tatgacaaga ttgagtttca ggggctggca
     2161 ccgattgctc gtgatttcta cgtctttatt ccctcctggc tatggcctga ccgtccgaat
     2221 ctggtgttaa acagcgcgaa ctatttcacc tgggaagtgt tgaataacca ctctgggctg
     2281 gcgatatccc ctacgctgtt aggctcgctg gtggtgatgg gcggcgtgct atttatcccg
     2341 ttgggtgcga ttgcggtagg gctggttatc aagtggttcg actgggtgta cgagttaggg
     2401 aagaacgaca gcaatcgcta taaggccgcc attttgcagg cgttctgctt cggtgcggtg
     2461 ttcaatatta ttgtgctaac gcgcgaaggc gtcgattcct ttgtttcacg tgtggtgttt
     2521 ttctgtctga tatttggcct gtgcctgctc gttgccaaac tgctttactg gctgctggaa
     2581 agcgccggac tcatccggca gcgtctgagg cgcatgcgcg ccacgccgct cgcgccgaca
     2641 cccaataccg tacaggatcc ggtgataaag gagcagatac tcgaggaaaa cctgtatttt
     2701 cagggcgcgt ggtcccatcc tcagttcgag aaaggcgcgg gttcaggtgc tggcagtgga
     2761 gcgtccgcgt ggagtcatcc gcaattcgaa aaatgagatc cggctgctaa caaagcccga
     2821 aaggaagctg agttggctgc tgccaccgct gagcaataac tagcataacc ccttggggcc
     2881 tctaaacggg tcttgagggg ttttttgctg aaaggaggaa ctatatccgg attggcgaat
     2941 gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     3001 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     3061 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     3121 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     3181 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     3241 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     3301 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     3361 ttaacgcgaa ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa
     3421 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     3481 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     3541 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     3601 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     3661 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     3721 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     3781 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     3841 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     3901 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     3961 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     4021 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgcagcaat
     4081 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     4141 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     4201 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     4261 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     4321 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     4381 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     4441 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     4501 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     4561 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     4621 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     4681 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
     4741 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     4801 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     4861 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     4921 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     4981 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     5041 gcttccaggg ggaaacgcct ggta
//
