LOCUS       sequence-339565-        4589 bp    DNA     circular SYN 15-APR-2025
DEFINITION  pQE30 encoding Wzz mutant  i128.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4589)
  AUTHORS   Papadopoulos M, Morona R
  TITLE     Mutagenesis and chemical cross-linking suggest that Wzz dimer 
            stability and oligomerization affect lipopolysaccharide O-antigen 
            modal chain length control.
  JOURNAL   J Bacteriol. 2010 Jul;192(13):3385-93. doi: 10.1128/JB.01134-09. 
            Epub 2010 May 7.
  PUBMED    20453100
REFERENCE   2  (bases 1 to 4589)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1128/JB.01134-09"; journalName: "J Bacteriol"; date: "2010-07"; 
            volume: "192"; issue: "13"; pages: "3385-93"
COMMENT     Sequence Label: pMPRMA39
FEATURES             Location/Qualifiers
     source          1..4589
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     12..30
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     protein_bind    74..90
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             109..120
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             139..156
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      1380..1474
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1518..2177
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1584..1603)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2242..2328
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(2375..2397)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2483..2623
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2538..2557
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2638..2655)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2809..3397)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2889..2908)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3568..4359,4360..4428))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4191..4210
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4429..4533)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 taacctataa aaataggcgt atcacgaggc cctttcgtct tcacctcgag aaatcataaa
       61 aaatttattt gctttgtgag cggataacaa tttcacacag aattcattaa agaggagaaa
      121 ttaactatga gaggatcgca tcaccatcac catcacggat ccgcatgcga gctcagagta
      181 gaaaataata atgtttctgg gcaaaaccat gacccggaac agattgattt gattgattta
      241 ctagtgcagt tgtggcgtgg caagatgaca attatcattt ccgtcattgt ggctattgcc
      301 ctggctattg gttatttggc agtagcgaag gagaaatgga cgtcaacagc aattatcact
      361 cagcccgacg tggggcaaat tgctggctat aacaatgcca tgaatgttat ctatggtcag
      421 gctgcaccga aagtatcgga tttgcaggag acgttaattg gtcgcttcag ttctgccttc
      481 tctgcattag cagaaacgct ggataatcag gaagagccag aaaaacttac catcgaacct
      541 tctgttaaga accagctgcg gccgcaccag caattaccat tgactgtttc ttatgttggg
      601 caaactgcag agggcgcaca aatgaagttg gcccaataca ttcagcaagt tgatgataaa
      661 gtgaatcaag agctagaaaa ggatctcaag gacaacattg ctctgggacg gaaaaacttg
      721 caggactctt taagaaccca ggaagtggtc gcgcaggagc agaaagatct gcgtatccgt
      781 cagattcagg aagcgttgca gtatgcgaat caggcgcagg tgacaaagcc acaggttcag
      841 cagactgaag atgtgacgca agatacgttg ttccttctag ggagcgaagc gctggagtcg
      901 atgattaagc atgaagcgac tcgtccgttg gtgttctcac caaactacta tcagacacgt
      961 caaaacctgt tggatattga aaaattaaag tttgatgatc ttgatattca tgcttaccgc
     1021 tatgtgatga aaccgacgtt acctattcgt cgcgatagtc cgaaaaaggc aatcaccttg
     1081 attctggcag tgcttctggg cggcatggtt ggcgcgggga ttgtgttggg gcgtaacgct
     1141 ctgcgtaatt acaacgcgaa gtaatattat tgtgcattta agagaaacgg gcagggtggt
     1201 gacaccatgc ccgttttttt tgccggatgc gatgctggcg catcttatcc ggcctacgtg
     1261 tgttgagata atgtgtaggc acgataagtt tgcgcatcgg gcaatggctc cgggtgtgac
     1321 aacaacatca cacctgctcc ccgggtcgac ctgcagccaa gcttaattag ctgagcttgg
     1381 actcctgttg atagatccag taatgacctc agaactccat ctggatttgt tcagaacgct
     1441 cggttgccgc cgggcgtttt ttattggtga gaatccaagc tagcttggcg agattttcag
     1501 gagctaagga agctaaaatg gagaaaaaaa tcactggata taccaccgtt gatatatccc
     1561 aatggcatcg taaagaacat tttgaggcat ttcagtcagt tgctcaatgt acctataacc
     1621 agaccgttca gctggatatt acggcctttt taaagaccgt aaagaaaaat aagcacaagt
     1681 tttatccggc ctttattcac attcttgccc gcctgatgaa tgctcatccg gaatttcgta
     1741 tggcaatgaa agacggtgag ctggtgatat gggatagtgt tcacccttgt tacaccgttt
     1801 tccatgagca aactgaaacg ttttcatcgc tctggagtga ataccacgac gatttccggc
     1861 agtttctaca catatattcg caagatgtgg cgtgttacgg tgaaaacctg gcctatttcc
     1921 ctaaagggtt tattgagaat atgtttttcg tctcagccaa tccctgggtg agtttcacca
     1981 gttttgattt aaacgtggcc aatatggaca acttcttcgc ccccgttttc accatgggca
     2041 aatattatac gcaaggcgac aaggtgctga tgccgctggc gattcaggtt catcatgccg
     2101 tttgtgatgg cttccatgtc ggcagaatgc ttaatgaatt acaacagtac tgcgatgagt
     2161 ggcagggcgg ggcgtaattt ttttaaggca gttattggtg cccttaaacg cctggggtaa
     2221 tgactctcta gcttgaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt
     2281 cgttttatct gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc gccctctaga
     2341 gctgcctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga
     2401 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag
     2461 cgggtgttgg cgggtgtcgg ggcgcagcca tgacccagtc acgtagcgat agcggagtgt
     2521 atactggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg
     2581 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgctctt ccgcttcctc
     2641 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     2701 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     2761 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     2821 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     2881 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     2941 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     3001 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     3061 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     3121 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     3181 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     3241 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     3301 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     3361 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     3421 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     3481 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     3541 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     3601 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     3661 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     3721 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     3781 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     3841 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     3901 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     3961 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     4021 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     4081 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     4141 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     4201 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     4261 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     4321 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     4381 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     4441 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     4501 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     4561 cgtctaagaa accattatta tcatgacat
//
