LOCUS       sequence-251146-        5681 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes MqsR toxin. Gent-R.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5681)
  AUTHORS   Lazarus JE, Warr AR, Kuehl CJ, Giorgio RT, Davis BM, Waldor MK
  TITLE     A New Suite of Allelic Exchange Vectors for the Scarless 
            Modification of Proteobacterial Genomes.
  JOURNAL   Appl Environ Microbiol. 2019 Jun 14. pii: AEM.00990-19. doi: 
            10.1128/AEM.00990-19.
  PUBMED    31201277
REFERENCE   2  (bases 1 to 5681)
  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: "Appl 
            Environ Microbiol. 2019 Jun 14. pii: AEM.00990-19. doi: 
            10.1128/AEM.00990-19."
COMMENT     Sequence Label: pTOX11
FEATURES             Location/Qualifiers
     source          1..5681
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(1..28)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(120..206)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             423..956
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     RBS             2096..2104
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     primer_bind     2268..2290
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2429..2448
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     oriT            3129..3238
                     /note="incP origin of transfer"
     CDS             3271..3642
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     rep_origin      4260..4648
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     protein_bind    4683..4699
                     /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)."
     terminator      complement(4735..4981)
                     /label=lambda tL3 terminator
                     /note="transcription terminator tL3 from phage lambda"
     promoter        complement(5361..5479)
                     /label=rhaB promoter
                     /note="promoter of the E. coli rhaBAD operon, conferring 
                     tight induction with L-rhamnose and repression with 
                     D-glucose in the presence of RhaR and RhaS (Giacalone et 
                     al., 2006)"
     regulatory      5619..5628
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 aaaaggccat ccgtcaggat ggccttctgc ttaatttgat gcctggcagt ttatggcggg
       61 cgtcctgccc gccaccctcc gggccgttgc ttcgcaacgt tcaaatccgc tcccggcgga
      121 tttgtcctac tcaggagagc gttcaccgac aaacaacaga taaaacgaaa ggcccagtct
      181 ttcgactgag cctttcgttt tatttgatgc ctggcagttc cctactctcg catggggaga
      241 ccccacacta ccatcggggg ccatcgatgc aacgggaatt tgaagacaat aactgcctta
      301 aaaaaattac gccccgccct gccactcatc gcagtactgt cgaatccatg tgggagttta
      361 ttcttgacac agatatttat gatataataa ctgagtaagc ttaacataag gaggaaaaac
      421 atatgttacg cagcagcaac gatgttacgc agcagggcag tcgccctaaa acaaagttag
      481 gtggctcaag tatgggcatc attcgcacat gtaggctcgg ccctgaccaa gtcaaatcca
      541 tgcgggctgc tcttgatctt ttcggtcgtg agttcggaga cgtagccacc tactcccaac
      601 atcagccgga ctccgattac ctcgggaact tgctccgtag taagacattc atcgcgcttg
      661 ctgccttcga ccaagaagcg gttgttggcg ctctcgcggc ttacgttctg cccaagtttg
      721 agcagccgcg tagtgagatc tatatctatg atctcgcagt ctccggcgag caccggaggc
      781 agggcattgc caccgcgctc atcaatctcc tcaagcatga ggccaacgcg cttggtgctt
      841 atgtgatcta cgtgcaagca gattacggtg acgatcccgc agtggctctc tatacaaagt
      901 tgggcatacg ggaagaagtg atgcactttg atatcgaccc aagtaccgcc acctaaagat
      961 ccttggcggc aagaaaacag gacacttggt atacgtgcta gcggccgcgg tccgatcgag
     1021 ctcgagacgt cccgggatgc atttaaatta attaacctgg tttaaacctt aagacgtcct
     1081 atcctagggc cctgtacacc atgtgcaccg gttcgaagat atcgtacgtg atcagcgaaa
     1141 aaaccccgcc gaagcggggt tttttgcgcg attagctcga agctcatgcg gtaccctctc
     1201 atccgccaaa acagccaagc ttgcatgcct gcaggtcgac tctagaaggc cttcactgta
     1261 agaaaccgtc gcggccttgg cggatgtcgc gagggctcca attgaattcg cgacggaaca
     1321 gggtgctgaa atgattgcta tccgaaaagc cacagtggta cgcaatatcg gtcaccgacg
     1381 cttccgaatg gcgcagtaag tggcgcgcct tcattaagcg taaacggttt aaatagcgtt
     1441 gcggggttaa gccggtctgt tgtttcagtt ggcggtgcag ggtgcgtaag ctcaggctga
     1501 actggtcggc aaccgcatcc cagttaactt cgtccgcgaa gtgatcttct aaccacgcta
     1561 acagcagatt caggcggctc gccgaattct ctaaattttc ttgcaggctc gacttgcgta
     1621 acagcagtaa cagctgcatg aacagaattt cacggctcgc cgtgctcggc aggtcgttct
     1681 cgccctcttg ttgctccatt tgcgccacta attggcgaac ttgttgcagc accgaatgat
     1741 tcacacgcca atggctaggg tattggccgt ctaattcctg cggtaacagt tggtttaagc
     1801 ccgccaggaa ttggaagcgg tcagggctac ggtataaaac gttggttaag cataagttgt
     1861 ccgtgtgctc gtataagtgg cggtcgtgat cacgaacgaa gcacacggta ccgcccgtaa
     1921 tcgtgtacgg ttggccgttg aaaacgtgga tgccggtacc gtgctccacg ataacgatct
     1981 cgtggaagtc gtggtggtgc tccgggaagt cggcttgagg caggcgcggc tcaatggcaa
     2041 cgctggcatt gccgctaggg aagaagtcaa ccgagtgcag caccgtcatt aaaaaacctc
     2101 cttatcggtg ataagctcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     2161 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgaggc
     2221 ctttacctta cttacggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     2281 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagcaaggag atggcgccca
     2341 acagtccccc ggccacgggg cctgccacca tacccacgcc gaaacaagcg ctcatgagcc
     2401 cgaagtggcg agcccgatct tccccatcgg tgatgtcggc gatataggcg ccagcaaccg
     2461 cacctgtggc gccggtgatg ccggccacga tgcgtccggc gtagaggatc ctttttgtcc
     2521 ggtgttgggt tgaaggtgaa gccggtcggg gccgcagcgg gggccggctt ttcagccttg
     2581 cccccctgct tcggccgccg tggctccggc gtcttgggtg ccggcgcggg ttccgcagcc
     2641 ttggcctgcg gtgcgggcac atcggcgggc ttggccttga tgtgccgcct ggcgtgcgag
     2701 cggaacgtct cgtaggagaa cttgaccttc cccgtttccc gcatgtgctc ccaaatggtg
     2761 acgagcgcat agccggacgc taacgccgcc tcgacatccg ccctcaccgc caggaacgca
     2821 accgcagcct catcacgccg gcgcttcttg gccgcgcggg attcaaccca ctcggccagc
     2881 tcgtcggtgt agctctttgg catcgtctct cgcctgtccc ctcagttcag taatttcctg
     2941 catttgcctg tttccagtcg gtagatattc cacaaaacag cagggaagca gcgcttttcc
     3001 gctgcataac cctgcttcgg ggtcattata gcgatttttt cggtatatcc atcctttttc
     3061 gcacgatata caggattttg ccaaagggtt cgtgtagact ttccttggtg tatccaacgg
     3121 cgtcagccgg gcaggatagg tgaagtaggc ccacccgcga gcgggtgttc cttcttcact
     3181 gtcccttatt cgcacctggc ggtgctcaac gggaatcctg ctctgcgagg ctggccggct
     3241 accgccggcg taacagatga gggcaagcgg atggctgatg aaaccaagcc aaccaggaag
     3301 ggcagcccac ctatcaaggt gtactgcctt ccagacgaac gaagagcgat tgaggaaaag
     3361 gcggcggcgg ccggcatgag cctgtcggcc tacctgctgg ccgtcggcca gggctacaaa
     3421 atcacgggcg tcgtggacta tgagcacgtc cgcgagctgg cccgcatcaa tggcgacctg
     3481 ggccgcctgg gcggcctgct gaaactctgg ctcaccgacg acccgcgcac ggcgcggttc
     3541 ggtgatgcca cgatcctcgc cctgctggcg aagatcgaag agaagcagga cgagcttggc
     3601 aaggtcatga tgggcgtggt ccgcccgagg gcagagccat gactttttta gccgctaaaa
     3661 cggccggggg gtgcgcgtga ttgccaagca cgtccccatg cgctccatca agaagagcga
     3721 cttcgcggag ctggtgaagt acatcaccga cgagcaaggc aagaccgagc gcctgggtca
     3781 cgtgcgcgtc acgaactgcg aggcaaacac cctgcccgct gtcatggccg aggtgatggc
     3841 gacccagcac ggcaacaccc gttccgaggc cgacaagacc tatcacctgc tggttagctt
     3901 ccgcgcggga gagaagcccg acgcggagac gttgcgcgcg attgaggacc gcatctgcgc
     3961 tgggcttggc ttcgccgagc atcagcgcgt cagtgccgtg catcacgaca ccgacaacct
     4021 gcacatccat atcgccatca acaagattca cccgacccga aacaccatcc atgagccgta
     4081 tcgggcctac cgcgccctcg ctgacctctg cgcgacgctc gaacgggact acgggcttga
     4141 gcgtgacaat cacgaaacgc ggcagcgcgt ttccgagaac cgcgcgaacg acatggagcg
     4201 gcacgcgggc gtggaaagcc tggtcggctg gatccggcca cgatgcgtcc ggcgtagagg
     4261 atctgaagat cagcagttca acctgttgat agtacgtact aagctctcat gtttcacgta
     4321 ctaagctctc atgtttaacg tactaagctc tcatgtttaa cgaactaaac cctcatggct
     4381 aacgtactaa gctctcatgg ctaacgtact aagctctcat gtttcacgta ctaagctctc
     4441 atgtttgaac aataaaatta atataaatca gcaacttaaa tagcctctaa ggttttaagt
     4501 tttataagaa aaaaaagaat atataaggct tttaaagctt ttaaggttta acggttgtgg
     4561 acaacaagcc agggatgtaa cgcactgaga agcccttaga gcctctcaaa gcaattttga
     4621 gtgacacagg aacacttaac ggctgacatg ggaattccac atgtggaatt ccacatgtgg
     4681 aattgtgagc ggataacaat ttgtggaatt cccccacgac ttcttcgtct gtttctactg
     4741 gtattggcac aaacctgatt ccaatttgag caaggctatg tgccatctcg atactcgttc
     4801 ttaactcaac agaagatgct ttgtgcatac agcccctcgt ttattattta tctcctcagc
     4861 cagccgctgt gctttcagtg gatttcggat aacagaaagg ccgggaaata cccagcctcg
     4921 ctttgtaacg gagtagagac gaaagtgatt gcgcctaccc ggatattatc gtgaggatgc
     4981 gtcatcgcca ttaattcact gatcagtgat aagctgtcaa acatgagaat tcgcggatcc
     5041 ttacttctcc ttaaacgaga cgatcagtac gtcatgaatt accgtaattt taagataaac
     5101 ctggcctgta acaagcctgg gtctgtaaac atcctgccag atagtatgat cagagtaggt
     5161 ggtcatgctt ttataaaagt cgctctctga taatccaatg ataacattac acataccatc
     5221 aaaatccaaa cctaactcat ctgcatttaa cagggcacta cgtgttgtac gaacttgccc
     5281 ggcattgaca agttttttaa cctgactcaa acgtgtatgt ggtgtgcgtt tttccatatg
     5341 gtgatcctgc tgaatttcat tacgaccagt ctaaaaagcg cctgaattcg cgaccttctc
     5401 gttactgaca ggaaaatggg ccattggcaa ccagggaaag atgaacgtga tgatgttcac
     5461 aatttgctga attgtggtga tgtgatgctc accgcatttc ctgaaaattc acgctgtatc
     5521 ttgaaaaatc gacgtttttt acgtggtttt ccgtcgaaaa tttaaggtaa gaacctgacc
     5581 tcgtgattac tatttcgccg tgttgacgac atcaggaggc caccatggcc tcgactagtt
     5641 ccggacagct gcaggcatgc aagcttggca ctggccacgc a
//
