LOCUS       sequence-345221-        4606 bp    DNA     circular SYN 15-APR-2025
DEFINITION  pQE30 with S. flexneri 2a wzzBSF gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4606)
  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 4606)
  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: pRMCD30
FEATURES             Location/Qualifiers
     source          1..4606
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(114..136)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    222..362
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     277..296
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(377..394)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(548..1136)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(628..647)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1307..2098,2099..2167))
                     /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     1930..1949
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2168..2272)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2340..2358
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(2382..2396,2397..2402,2403..2419,2420..2426)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    2402..2418
                     /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    2434..2450
                     /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             2469..2480
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             2499..2516
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      3725..3819
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             3863..4522
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3929..3948)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      join(4587..4606,1..67)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag
       61 gacaaatccg ccctctagag ctgcctcgcg cgtttcggtg atgacggtga aaacctctga
      121 cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa
      181 gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat gacccagtca
      241 cgtagcgata gcggagtgta tactggctta actatgcggc atcagagcag attgtactga
      301 gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca
      361 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
      421 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
      481 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
      541 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
      601 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
      661 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
      721 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
      781 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
      841 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
      901 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
      961 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
     1021 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     1081 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     1141 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     1201 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     1261 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     1321 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     1381 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     1441 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     1501 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     1561 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     1621 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     1681 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     1741 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     1801 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     1861 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     1921 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     1981 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     2041 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     2101 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     2161 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     2221 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     2281 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     2341 ataggcgtat cacgaggccc tttcgtcttc acctcgagaa atcataaaaa atttatttgc
     2401 tttgtgagcg gataacaatt ataatagatt caattgtgag cggataacaa tttcacacag
     2461 aattcattaa agaggagaaa ttaactatga gaggatcgca tcaccatcac catcacggat
     2521 ccgcatgcga gctcagagta gaaaataata atgtttctgg gcaaaaccat gacccggaac
     2581 agattgattt gattgattta ctagtgcagt tgtggcgtgg caagatgaca attatcattt
     2641 ccgtcattgt ggctattgcc ctggctattg gttatttggc agtagcgaag gagaaatgga
     2701 cgtcaacagc aattatcact cagcccgacg tggggcaaat tgctggctat aacaatgcca
     2761 tgaatgttat ctatggtcag gctgcaccga aagtatcgga tttgcaggag acgttaattg
     2821 gtcgcttcag ttctgccttc tctgcattag cagaaacgct ggataatcag gaagagccag
     2881 aaaaacttac catcgaacct tctgttaaga accagcaatt accattgact gtttcttatg
     2941 ttgggcaaac tgcagagggc gcacaaatga agttggccca atacattcag caagttgatg
     3001 ataaagtgaa tcaagagcta gaaaaggatc tcaaggacaa cattgctctg ggacggaaaa
     3061 acttgcagga ctctttaaga acccaggaag tggtcgcgca ggagcagaaa gatctgcgta
     3121 tccgtcagat tcaggaagcg ttgcagtatg cgaatcaggc gcaggtgaca aagccacagg
     3181 ttcagcagac tgaagatgtg acgcaagata cgttgttcct tctagggagc gaagcgctgg
     3241 agtcgatgat taagcatgaa gcgactcgtc cgttggtgtt ctcaccaaac tactatcaga
     3301 cacgtcaaaa cctgttggat attgaaaaat taaagtttga tgatcttgat attcatgctt
     3361 accgctatgt gatgaaaccg acgttaccta ttcgtcgcga tagtccgaaa aaggcaatca
     3421 ccttgattct ggcagtgctt ctgggcggca tggttggcgc ggggattgtg ttggggcgta
     3481 acgctctgcg taattacaac gcgaagtaat attattgtgc atttaagaga aacgggcagg
     3541 gtggtgacac catgcccgtt ttttttgccg gatgcgatgc tggcgcatct tatccggcct
     3601 acgtgtgttg agataatgtg taggcacgat aagtttgcgc atcgggcaat ggctccgggt
     3661 gtgacaacaa catcacacct gctccccggg tcgacctgca gccaagctta attagctgag
     3721 cttggactcc tgttgataga tccagtaatg acctcagaac tccatctgga tttgttcaga
     3781 acgctcggtt gccgccgggc gttttttatt ggtgagaatc caagctagct tggcgagatt
     3841 ttcaggagct aaggaagcta aaatggagaa aaaaatcact ggatatacca ccgttgatat
     3901 atcccaatgg catcgtaaag aacattttga ggcatttcag tcagttgctc aatgtaccta
     3961 taaccagacc gttcagctgg atattacggc ctttttaaag accgtaaaga aaaataagca
     4021 caagttttat ccggccttta ttcacattct tgcccgcctg atgaatgctc atccggaatt
     4081 tcgtatggca atgaaagacg gtgagctggt gatatgggat agtgttcacc cttgttacac
     4141 cgttttccat gagcaaactg aaacgttttc atcgctctgg agtgaatacc acgacgattt
     4201 ccggcagttt ctacacatat attcgcaaga tgtggcgtgt tacggtgaaa acctggccta
     4261 tttccctaaa gggtttattg agaatatgtt tttcgtctca gccaatccct gggtgagttt
     4321 caccagtttt gatttaaacg tggccaatat ggacaacttc ttcgcccccg ttttcaccat
     4381 gggcaaatat tatacgcaag gcgacaaggt gctgatgccg ctggcgattc aggttcatca
     4441 tgccgtttgt gatggcttcc atgtcggcag aatgcttaat gaattacaac agtactgcga
     4501 tgagtggcag ggcggggcgt aattttttta aggcagttat tggtgccctt aaacgcctgg
     4561 ggtaatgact ctctagcttg aggcatcaaa taaaacgaaa ggctca
//
