LOCUS       sequence-345223-        4621 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pQE30 encoding Wzz mutant  i92.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4621)
  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 4621)
  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"; doi: 
            "10.1128/JB.01134-09"; journalName: "J Bacteriol"; date: "2010-07"; 
            volume: "192"; issue: "13"; pages: "3385-93"
COMMENT     Sequence Label: pMPRMA38
FEATURES             Location/Qualifiers
     source          1..4621
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     33..51
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(75..89,90..95,96..112,113..119)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    95..111
                     /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)."
     protein_bind    127..143
                     /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             162..173
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             192..209
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      1433..1527
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1571..2230
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1637..1656)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2295..2381
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(2428..2450)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2536..2676
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2591..2610
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2691..2708)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2862..3450)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2942..2961)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3621..4412,4413..4481))
                     /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     4244..4263
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4482..4586)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
       61 ttcacctcga gaaatcataa aaaatttatt tgctttgtga gcggataaca attataatag
      121 attcaattgt gagcggataa caatttcaca cagaattcat taaagaggag aaattaacta
      181 tgagaggatc gcatcaccat caccatcacg gatccgcatg cgagctcaga gtagaaaata
      241 ataatgtttc tgggcaaaac catgacccgg aacagattga tttgattgat ttactagtgc
      301 agttgtggcg tggcaagatg acaattatca tttccgtcat tgtggctatt gccctggcta
      361 ttggttattt ggcagtagcg aaggagaaat ggacgtcaac agcaattatc actcagcccg
      421 acgtggggca aattgctggc tataacaatg ccatgaatgt tatctatggt caggctgcac
      481 cgaaagtatc ggatttgcag gtgcggccgc agcaggagac gttaattggt cgcttcagtt
      541 ctgccttctc tgcattagca gaaacgctgg ataatcagga agagccagaa aaacttacca
      601 tcgaaccttc tgttaagaac cagcaattac cattgactgt ttcttatgtt gggcaaactg
      661 cagagggcgc acaaatgaag ttggcccaat acattcagca agttgatgat aaagtgaatc
      721 aagagctaga aaaggatctc aaggacaaca ttgctctggg acggaaaaac ttgcaggact
      781 ctttaagaac ccaggaagtg gtcgcgcagg agcagaaaga tctgcgtatc cgtcagattc
      841 aggaagcgtt gcagtatgcg aatcaggcgc aggtgacaaa gccacaggtt cagcagactg
      901 aagatgtgac gcaagatacg ttgttccttc tagggagcga agcgctggag tcgatgatta
      961 agcatgaagc gactcgtccg ttggtgttct caccaaacta ctatcagaca cgtcaaaacc
     1021 tgttggatat tgaaaaatta aagtttgatg atcttgatat tcatgcttac cgctatgtga
     1081 tgaaaccgac gttacctatt cgtcgcgata gtccgaaaaa ggcaatcacc ttgattctgg
     1141 cagtgcttct gggcggcatg gttggcgcgg ggattgtgtt ggggcgtaac gctctgcgta
     1201 attacaacgc gaagtaatat tattgtgcat ttaagagaaa cgggcagggt ggtgacacca
     1261 tgcccgtttt ttttgccgga tgcgatgctg gcgcatctta tccggcctac gtgtgttgag
     1321 ataatgtgta ggcacgataa gtttgcgcat cgggcaatgg ctccgggtgt gacaacaaca
     1381 tcacacctgc tccccgggtc gacctgcagc caagcttaat tagctgagct tggactcctg
     1441 ttgatagatc cagtaatgac ctcagaactc catctggatt tgttcagaac gctcggttgc
     1501 cgccgggcgt tttttattgg tgagaatcca agctagcttg gcgagatttt caggagctaa
     1561 ggaagctaaa atggagaaaa aaatcactgg atataccacc gttgatatat cccaatggca
     1621 tcgtaaagaa cattttgagg catttcagtc agttgctcaa tgtacctata accagaccgt
     1681 tcagctggat attacggcct ttttaaagac cgtaaagaaa aataagcaca agttttatcc
     1741 ggcctttatt cacattcttg cccgcctgat gaatgctcat ccggaatttc gtatggcaat
     1801 gaaagacggt gagctggtga tatgggatag tgttcaccct tgttacaccg ttttccatga
     1861 gcaaactgaa acgttttcat cgctctggag tgaataccac gacgatttcc ggcagtttct
     1921 acacatatat tcgcaagatg tggcgtgtta cggtgaaaac ctggcctatt tccctaaagg
     1981 gtttattgag aatatgtttt tcgtctcagc caatccctgg gtgagtttca ccagttttga
     2041 tttaaacgtg gccaatatgg acaacttctt cgcccccgtt ttcaccatgg gcaaatatta
     2101 tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag gttcatcatg ccgtttgtga
     2161 tggcttccat gtcggcagaa tgcttaatga attacaacag tactgcgatg agtggcaggg
     2221 cggggcgtaa tttttttaag gcagttattg gtgcccttaa acgcctgggg taatgactct
     2281 ctagcttgag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta
     2341 tctgttgttt gtcggtgaac gctctcctga gtaggacaaa tccgccctct agagctgcct
     2401 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
     2461 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
     2521 tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc gatagcggag tgtatactgg
     2581 cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata
     2641 ccgcacagat gcgtaaggag aaaataccgc atcaggcgct cttccgcttc ctcgctcact
     2701 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     2761 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     2821 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     2881 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     2941 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     3001 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     3061 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     3121 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     3181 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     3241 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     3301 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     3361 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     3421 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     3481 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     3541 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     3601 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     3661 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     3721 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     3781 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     3841 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     3901 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     3961 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     4021 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     4081 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     4141 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     4201 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     4261 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     4321 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     4381 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     4441 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     4501 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     4561 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     4621 g
//
