LOCUS       sequence-340808-        4589 bp    DNA     circular SYN 15-APR-2025
DEFINITION  pQE30 encoding Wzz mutant  i290.
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: pMPRMA42
FEATURES             Location/Qualifiers
     source          1..4589
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      1205..1299
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1343..2002
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1409..1428)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2067..2153
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(2200..2222)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2308..2448
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2363..2382
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2463..2480)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2634..3222)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2714..2733)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3393..4184,4185..4253))
                     /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     4016..4035
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4254..4358)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4426..4444
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     protein_bind    4488..4504
                     /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             4523..4534
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             4553..4570
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
ORIGIN
        1 gagtagaaaa taataatgtt tctgggcaaa accatgaccc ggaacagatt gatttgattg
       61 atttactagt gcagttgtgg cgtggcaaga tgacaattat catttccgtc attgtggcta
      121 ttgccctggc tattggttat ttggcagtag cgaaggagaa atggacgtca acagcaatta
      181 tcactcagcc cgacgtgggg caaattgctg gctataacaa tgccatgaat gttatctatg
      241 gtcaggctgc accgaaagta tcggatttgc aggagacgtt aattggtcgc ttcagttctg
      301 ccttctctgc attagcagaa acgctggata atcaggaaga gccagaaaaa cttaccatcg
      361 aaccttctgt taagaaccag caattaccat tgactgtttc ttatgttggg caaactgcag
      421 agggcgcaca aatgaagttg gcccaataca ttcagcaagt tgatgataaa gtgaatcaag
      481 agctagaaaa ggatctcaag gacaacattg ctctgggacg gaaaaacttg caggactctt
      541 taagaaccca ggaagtggtc gcgcaggagc agaaagatct gcgtatccgt cagattcagg
      601 aagcgttgca gtatgcgaat caggcgcagg tgacaaagcc acaggttcag cagactgaag
      661 atgtgacgca agatacgttg ttccttctag ggagcgaagc gctggagtcg atgattaagc
      721 atgaagcgac tcgtccgttg gtgttctcac caaactacta tcagacacgt caaaacctgt
      781 tggatattga aaaattaaag tttgatgatc ttgatattca tgcttaccgc tatgtgatga
      841 aaccgacgtt acctattcgt cgcgattgcg gccgcagcga tagtccgaaa aaggcaatca
      901 ccttgattct ggcagtgctt ctgggcggca tggttggcgc ggggattgtg ttggggcgta
      961 acgctctgcg taattacaac gcgaagtaat attattgtgc atttaagaga aacgggcagg
     1021 gtggtgacac catgcccgtt ttttttgccg gatgcgatgc tggcgcatct tatccggcct
     1081 acgtgtgttg agataatgtg taggcacgat aagtttgcgc atcgggcaat ggctccgggt
     1141 gtgacaacaa catcacacct gctccccggg tcgacctgca gccaagctta attagctgag
     1201 cttggactcc tgttgataga tccagtaatg acctcagaac tccatctgga tttgttcaga
     1261 acgctcggtt gccgccgggc gttttttatt ggtgagaatc caagctagct tggcgagatt
     1321 ttcaggagct aaggaagcta aaatggagaa aaaaatcact ggatatacca ccgttgatat
     1381 atcccaatgg catcgtaaag aacattttga ggcatttcag tcagttgctc aatgtaccta
     1441 taaccagacc gttcagctgg atattacggc ctttttaaag accgtaaaga aaaataagca
     1501 caagttttat ccggccttta ttcacattct tgcccgcctg atgaatgctc atccggaatt
     1561 tcgtatggca atgaaagacg gtgagctggt gatatgggat agtgttcacc cttgttacac
     1621 cgttttccat gagcaaactg aaacgttttc atcgctctgg agtgaatacc acgacgattt
     1681 ccggcagttt ctacacatat attcgcaaga tgtggcgtgt tacggtgaaa acctggccta
     1741 tttccctaaa gggtttattg agaatatgtt tttcgtctca gccaatccct gggtgagttt
     1801 caccagtttt gatttaaacg tggccaatat ggacaacttc ttcgcccccg ttttcaccat
     1861 gggcaaatat tatacgcaag gcgacaaggt gctgatgccg ctggcgattc aggttcatca
     1921 tgccgtttgt gatggcttcc atgtcggcag aatgcttaat gaattacaac agtactgcga
     1981 tgagtggcag ggcggggcgt aattttttta aggcagttat tggtgccctt aaacgcctgg
     2041 ggtaatgact ctctagcttg aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg
     2101 cctttcgttt tatctgttgt ttgtcggtga acgctctcct gagtaggaca aatccgccct
     2161 ctagagctgc ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
     2221 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
     2281 gtcagcgggt gttggcgggt gtcggggcgc agccatgacc cagtcacgta gcgatagcgg
     2341 agtgtatact ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg
     2401 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ctcttccgct
     2461 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     2521 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     2581 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     2641 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     2701 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     2761 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     2821 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     2881 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     2941 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     3001 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     3061 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     3121 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     3181 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     3241 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     3301 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     3361 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     3421 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     3481 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     3541 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     3601 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     3661 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     3721 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     3781 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     3841 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     3901 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     3961 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     4021 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     4081 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     4141 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     4201 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     4261 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     4321 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     4381 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     4441 aggccctttc gtcttcacct cgagaaatca taaaaaattt atttgctttg tgagcggata
     4501 acaatttcac acagaattca ttaaagagga gaaattaact atgagaggat cgcatcacca
     4561 tcaccatcac ggatccgcat gcgagctca
//
