LOCUS       sequence-251148-        6629 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes MqsR toxin and tsPurple chromoprotein. CAM-R.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6629)
  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 6629)
  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: pTOX8
FEATURES             Location/Qualifiers
     source          1..6629
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        326..428
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             429..1088
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(495..514)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1189..1275
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1367..1394
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     regulatory      1448..1457
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        1597..1715
                     /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)"
     terminator      2095..2341
                     /label=lambda tL3 terminator
                     /note="transcription terminator tL3 from phage lambda"
     protein_bind    2377..2393
                     /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)."
     rep_origin      2428..2816
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     CDS             complement(3434..3805)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            3838..3947
                     /note="incP origin of transfer"
     primer_bind     complement(4628..4647)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(4786..4808)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     RBS             4972..4980
                     /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)"
     RBS             5902..5913
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
ORIGIN
        1 ttggcggatg agagggtacc gcatgagctt cgagctaatc gcgcaaaaaa ccccgcttcg
       61 gcggggtttt ttcgctgatc acgtacgata tcttcgaacc ggtgcacatg gtgtacaggg
      121 ccctaggata ggacgtctta aggtttaaac caggttaatt aatttaaatg catcccggga
      181 cgtctcgagc tcgatcggac cgcggccgct agcacgtata ccaagtgtcc tgttttcttg
      241 ccgccaagga tctgatggcg caggggatca agatctctgt tgataccggg aagccctggg
      301 ccaacttttg gcgaaaatga gacgttgatc ggcacgtaag aggttccaac tttcaccata
      361 atgaaataag atcactaccg ggcgtatttt ttgagttatc gagattttca ggagctaagg
      421 aagctaaaat ggagaaaaaa atcactggat ataccaccgt tgatatatcc caatggcatc
      481 gtaaagaaca ttttgaggca tttcagtcag ttgctcaatg tacctataac cagaccgttc
      541 agctggatat tacggccttt ttaaagaccg taaagaaaaa taagcacaag ttttatccgg
      601 cctttattca cattcttgcc cgcctgatga atgctcatcc ggaattacgt atggcaatga
      661 aagacggtga gctggtgata tgggatagtg ttcacccttg ttacaccgtt ttccatgagc
      721 aaactgaaac gttttcatcg ctctggagtg aataccacga cgatttccgg cagtttctac
      781 acatatattc gcaagatgtg gcgtgttacg gtgaaaacct ggcctatttc cctaaagggt
      841 ttattgagaa tatgtttttc gtctcagcca atccctgggt gagtttcacc agttttgatt
      901 taaacgtggc caatatggac aacttcttcg cccccgtttt caccatgggc aaatattata
      961 cgcaaggcga caaggtgctg atgccgctgg cgattcaggt tcatcatgcc gtttgtgatg
     1021 gcttccatgt cggcagaatg cttaatgaat tacaacagta ctgcgatgag tggcagggcg
     1081 gggcgtaatt tttttaaggc agttattgtc ttcaaattcc cgttgcatcg atggcccccg
     1141 atggtagtgt ggggtctccc catgcgagag tagggaactg ccaggcatca aataaaacga
     1201 aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc
     1261 ctgagtagga caaatccgcc gggagcggat ttgaacgttg cgaagcaacg gcccggaggg
     1321 tggcgggcag gacgcccgcc ataaactgcc aggcatcaaa ttaagcagaa ggccatcctg
     1381 acggatggcc tttttgcgtg gccagtgcca agcttgcatg cctgcagctg tccggaacta
     1441 gtcgaggcca tggtggcctc ctgatgtcgt caacacggcg aaatagtaat cacgaggtca
     1501 ggttcttacc ttaaattttc gacggaaaac cacgtaaaaa acgtcgattt ttcaagatac
     1561 agcgtgaatt ttcaggaaat gcggtgagca tcacatcacc acaattcagc aaattgtgaa
     1621 catcatcacg ttcatctttc cctggttgcc aatggcccat tttcctgtca gtaacgagaa
     1681 ggtcgcgaat tcaggcgctt tttagactgg tcgtaatgaa attcagcagg atcaccatat
     1741 ggaaaaacgc acaccacata cacgtttgag tcaggttaaa aaacttgtca atgccgggca
     1801 agttcgtaca acacgtagtg ccctgttaaa tgcagatgag ttaggtttgg attttgatgg
     1861 tatgtgtaat gttatcattg gattatcaga gagcgacttt tataaaagca tgaccaccta
     1921 ctctgatcat actatctggc aggatgttta cagacccagg cttgttacag gccaggttta
     1981 tcttaaaatt acggtaattc atgacgtact gatcgtctcg tttaaggaga agtaaggatc
     2041 cgcgaattct catgtttgac agcttatcac tgatcagtga attaatggcg atgacgcatc
     2101 ctcacgataa tatccgggta ggcgcaatca ctttcgtctc tactccgtta caaagcgagg
     2161 ctgggtattt cccggccttt ctgttatccg aaatccactg aaagcacagc ggctggctga
     2221 ggagataaat aataaacgag gggctgtatg cacaaagcat cttctgttga gttaagaacg
     2281 agtatcgaga tggcacatag ccttgctcaa attggaatca ggtttgtgcc aataccagta
     2341 gaaacagacg aagaagtcgt gggggaattc cacaaattgt tatccgctca caattccaca
     2401 tgtggaattc cacatgtgga attcccatgt cagccgttaa gtgttcctgt gtcactcaaa
     2461 attgctttga gaggctctaa gggcttctca gtgcgttaca tccctggctt gttgtccaca
     2521 accgttaaac cttaaaagct ttaaaagcct tatatattct tttttttctt ataaaactta
     2581 aaaccttaga ggctatttaa gttgctgatt tatattaatt ttattgttca aacatgagag
     2641 cttagtacgt gaaacatgag agcttagtac gttagccatg agagcttagt acgttagcca
     2701 tgagggttta gttcgttaaa catgagagct tagtacgtta aacatgagag cttagtacgt
     2761 gaaacatgag agcttagtac gtactatcaa caggttgaac tgctgatctt cagatcctct
     2821 acgccggacg catcgtggcc ggatccagcc gaccaggctt tccacgcccg cgtgccgctc
     2881 catgtcgttc gcgcggttct cggaaacgcg ctgccgcgtt tcgtgattgt cacgctcaag
     2941 cccgtagtcc cgttcgagcg tcgcgcagag gtcagcgagg gcgcggtagg cccgatacgg
     3001 ctcatggatg gtgtttcggg tcgggtgaat cttgttgatg gcgatatgga tgtgcaggtt
     3061 gtcggtgtcg tgatgcacgg cactgacgcg ctgatgctcg gcgaagccaa gcccagcgca
     3121 gatgcggtcc tcaatcgcgc gcaacgtctc cgcgtcgggc ttctctcccg cgcggaagct
     3181 aaccagcagg tgataggtct tgtcggcctc ggaacgggtg ttgccgtgct gggtcgccat
     3241 cacctcggcc atgacagcgg gcagggtgtt tgcctcgcag ttcgtgacgc gcacgtgacc
     3301 caggcgctcg gtcttgcctt gctcgtcggt gatgtacttc accagctccg cgaagtcgct
     3361 cttcttgatg gagcgcatgg ggacgtgctt ggcaatcacg cgcacccccc ggccgtttta
     3421 gcggctaaaa aagtcatggc tctgccctcg ggcggaccac gcccatcatg accttgccaa
     3481 gctcgtcctg cttctcttcg atcttcgcca gcagggcgag gatcgtggca tcaccgaacc
     3541 gcgccgtgcg cgggtcgtcg gtgagccaga gtttcagcag gccgcccagg cggcccaggt
     3601 cgccattgat gcgggccagc tcgcggacgt gctcatagtc cacgacgccc gtgattttgt
     3661 agccctggcc gacggccagc aggtaggccg acaggctcat gccggccgcc gccgcctttt
     3721 cctcaatcgc tcttcgttcg tctggaaggc agtacacctt gataggtggg ctgcccttcc
     3781 tggttggctt ggtttcatca gccatccgct tgccctcatc tgttacgccg gcggtagccg
     3841 gccagcctcg cagagcagga ttcccgttga gcaccgccag gtgcgaataa gggacagtga
     3901 agaaggaaca cccgctcgcg ggtgggccta cttcacctat cctgcccggc tgacgccgtt
     3961 ggatacacca aggaaagtct acacgaaccc tttggcaaaa tcctgtatat cgtgcgaaaa
     4021 aggatggata taccgaaaaa atcgctataa tgaccccgaa gcagggttat gcagcggaaa
     4081 agcgctgctt ccctgctgtt ttgtggaata tctaccgact ggaaacaggc aaatgcagga
     4141 aattactgaa ctgaggggac aggcgagaga cgatgccaaa gagctacacc gacgagctgg
     4201 ccgagtgggt tgaatcccgc gcggccaaga agcgccggcg tgatgaggct gcggttgcgt
     4261 tcctggcggt gagggcggat gtcgaggcgg cgttagcgtc cggctatgcg ctcgtcacca
     4321 tttgggagca catgcgggaa acggggaagg tcaagttctc ctacgagacg ttccgctcgc
     4381 acgccaggcg gcacatcaag gccaagcccg ccgatgtgcc cgcaccgcag gccaaggctg
     4441 cggaacccgc gccggcaccc aagacgccgg agccacggcg gccgaagcag gggggcaagg
     4501 ctgaaaagcc ggcccccgct gcggccccga ccggcttcac cttcaaccca acaccggaca
     4561 aaaaggatcc tctacgccgg acgcatcgtg gccggcatca ccggcgccac aggtgcggtt
     4621 gctggcgcct atatcgccga catcaccgat ggggaagatc gggctcgcca cttcgggctc
     4681 atgagcgctt gtttcggcgt gggtatggtg gcaggccccg tggccggggg actgttgggc
     4741 gccatctcct tgctgcctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     4801 gctcccggag acggtcacag cttgtctgta agcggatgcc gtaagtaagg taaaggcctc
     4861 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     4921 aataaccctg ataaatgctt caataatatt gaaaaaggag cttatcaccg ataaggaggt
     4981 tttttaatga cggtgctgca ctcggttgac ttcttcccta gcggcaatgc cagcgttgcc
     5041 attgagccgc gcctgcctca agccgacttc ccggagcacc accacgactt ccacgagatc
     5101 gttatcgtgg agcacggtac cggcatccac gttttcaacg gccaaccgta cacgattacg
     5161 ggcggtaccg tgtgcttcgt tcgtgatcac gaccgccact tatacgagca cacggacaac
     5221 ttatgcttaa ccaacgtttt ataccgtagc cctgaccgct tccaattcct ggcgggctta
     5281 aaccaactgt taccgcagga attagacggc caatacccta gccattggcg tgtgaatcat
     5341 tcggtgctgc aacaagttcg ccaattagtg gcgcaaatgg agcaacaaga gggcgagaac
     5401 gacctgccga gcacggcgag ccgtgaaatt ctgttcatgc agctgttact gctgttacgc
     5461 aagtcgagcc tgcaagaaaa tttagagaat tcggcgagcc gcctgaatct gctgttagcg
     5521 tggttagaag atcacttcgc ggacgaagtt aactgggatg cggttgccga ccagttcagc
     5581 ctgagcttac gcaccctgca ccgccaactg aaacaacaga ccggcttaac cccgcaacgc
     5641 tatttaaacc gtttacgctt aatgaaggcg cgccacttac tgcgccattc ggaagcgtcg
     5701 gtgaccgata ttgcgtacca ctgtggcttt tcggatagca atcatttcag caccctgttc
     5761 cgtcgcgaat tcaattggag ccctcgcgac atccgccaag gccgcgacgg tttcttacag
     5821 tgaaggcctt ctagagtcga cctgcaaaaa agagtattga cttcgcatct ttttgtacct
     5881 ataatagatt cattactaga gaaagaggag aaatactaga tggcgtccct tgtaaagaag
     5941 gatatgtgtg ttaagatgac aatggaggga accgtcaacg ggtatcactt taagtgtgtt
     6001 ggcgagggag aaggcaaacc gttcgagggt acacagaata tgcgcattcg tgtcaccgaa
     6061 gggggccctt tgccctttgc tttcgatatt ttggccccgt gctgtatgta tggttcgaag
     6121 acttttatca aacacgtaag cggaatccct gactacttca aggagagctt tcccgaaggc
     6181 tttacttggg aacgtacgca gatttttgaa gacggaggag ttttgactgc ccatcaagat
     6241 acatcacttg agggaaactg tttaatctac aaagttaagg tgttagggac aaattttcct
     6301 gccaatggtc cggttatgca gaaaaagaca gcaggctggg aaccgtgtgt agagatgttg
     6361 tacccccgcg acggggttct ttgcgggcaa tcccttatgg ccttgaaatg tactgatgga
     6421 aatcacttga catcccattt acgcaccact tatcgctcgc gtaagccaag taatgcggtg
     6481 aacatgccgg aatttcactt cggagaccat cgtattgaaa tcttgaaagc cgaacaagga
     6541 aaattttatg aacaatacga atccgcagtg gcacgttatt cagatgttcc agaaaaggcg
     6601 acttgataag gcatgcaagc ttggctgtt
//
