LOCUS       sequence-249680-        6640 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes MqsR toxin and amilCP chromoprotein. CAM-R.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6640)
  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 6640)
  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: pTOX5
FEATURES             Location/Qualifiers
     source          1..6640
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..389
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     CDS             complement(1007..1378)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            1411..1520
                     /note="incP origin of transfer"
     primer_bind     complement(2201..2220)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(2359..2381)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     RBS             2545..2553
                     /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)"
     promoter        join(3431..3436,3437..3452,3453..3459)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     promoter        4539..4641
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             4642..5301
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(4708..4727)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      5402..5488
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5580..5607
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     regulatory      5661..5670
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        5810..5928
                     /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      6308..6554
                     /label=lambda tL3 terminator
                     /note="transcription terminator tL3 from phage lambda"
     protein_bind    6590..6606
                     /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)."
ORIGIN
        1 tgtcagccgt taagtgttcc tgtgtcactc aaaattgctt tgagaggctc taagggcttc
       61 tcagtgcgtt acatccctgg cttgttgtcc acaaccgtta aaccttaaaa gctttaaaag
      121 ccttatatat tctttttttt cttataaaac ttaaaacctt agaggctatt taagttgctg
      181 atttatatta attttattgt tcaaacatga gagcttagta cgtgaaacat gagagcttag
      241 tacgttagcc atgagagctt agtacgttag ccatgagggt ttagttcgtt aaacatgaga
      301 gcttagtacg ttaaacatga gagcttagta cgtgaaacat gagagcttag tacgtactat
      361 caacaggttg aactgctgat cttcagatcc tctacgccgg acgcatcgtg gccggatcca
      421 gccgaccagg ctttccacgc ccgcgtgccg ctccatgtcg ttcgcgcggt tctcggaaac
      481 gcgctgccgc gtttcgtgat tgtcacgctc aagcccgtag tcccgttcga gcgtcgcgca
      541 gaggtcagcg agggcgcggt aggcccgata cggctcatgg atggtgtttc gggtcgggtg
      601 aatcttgttg atggcgatat ggatgtgcag gttgtcggtg tcgtgatgca cggcactgac
      661 gcgctgatgc tcggcgaagc caagcccagc gcagatgcgg tcctcaatcg cgcgcaacgt
      721 ctccgcgtcg ggcttctctc ccgcgcggaa gctaaccagc aggtgatagg tcttgtcggc
      781 ctcggaacgg gtgttgccgt gctgggtcgc catcacctcg gccatgacag cgggcagggt
      841 gtttgcctcg cagttcgtga cgcgcacgtg acccaggcgc tcggtcttgc cttgctcgtc
      901 ggtgatgtac ttcaccagct ccgcgaagtc gctcttcttg atggagcgca tggggacgtg
      961 cttggcaatc acgcgcaccc cccggccgtt ttagcggcta aaaaagtcat ggctctgccc
     1021 tcgggcggac cacgcccatc atgaccttgc caagctcgtc ctgcttctct tcgatcttcg
     1081 ccagcagggc gaggatcgtg gcatcaccga accgcgccgt gcgcgggtcg tcggtgagcc
     1141 agagtttcag caggccgccc aggcggccca ggtcgccatt gatgcgggcc agctcgcgga
     1201 cgtgctcata gtccacgacg cccgtgattt tgtagccctg gccgacggcc agcaggtagg
     1261 ccgacaggct catgccggcc gccgccgcct tttcctcaat cgctcttcgt tcgtctggaa
     1321 ggcagtacac cttgataggt gggctgccct tcctggttgg cttggtttca tcagccatcc
     1381 gcttgccctc atctgttacg ccggcggtag ccggccagcc tcgcagagca ggattcccgt
     1441 tgagcaccgc caggtgcgaa taagggacag tgaagaagga acacccgctc gcgggtgggc
     1501 ctacttcacc tatcctgccc ggctgacgcc gttggataca ccaaggaaag tctacacgaa
     1561 ccctttggca aaatcctgta tatcgtgcga aaaaggatgg atataccgaa aaaatcgcta
     1621 taatgacccc gaagcagggt tatgcagcgg aaaagcgctg cttccctgct gttttgtgga
     1681 atatctaccg actggaaaca ggcaaatgca ggaaattact gaactgaggg gacaggcgag
     1741 agacgatgcc aaagagctac accgacgagc tggccgagtg ggttgaatcc cgcgcggcca
     1801 agaagcgccg gcgtgatgag gctgcggttg cgttcctggc ggtgagggcg gatgtcgagg
     1861 cggcgttagc gtccggctat gcgctcgtca ccatttggga gcacatgcgg gaaacgggga
     1921 aggtcaagtt ctcctacgag acgttccgct cgcacgccag gcggcacatc aaggccaagc
     1981 ccgccgatgt gcccgcaccg caggccaagg ctgcggaacc cgcgccggca cccaagacgc
     2041 cggagccacg gcggccgaag caggggggca aggctgaaaa gccggccccc gctgcggccc
     2101 cgaccggctt caccttcaac ccaacaccgg acaaaaagga tcctctacgc cggacgcatc
     2161 gtggccggca tcaccggcgc cacaggtgcg gttgctggcg cctatatcgc cgacatcacc
     2221 gatggggaag atcgggctcg ccacttcggg ctcatgagcg cttgtttcgg cgtgggtatg
     2281 gtggcaggcc ccgtggccgg gggactgttg ggcgccatct ccttgctgcc tcgcgcgttt
     2341 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     2401 gtaagcggat gccgtaagta aggtaaaggc ctcgcggaac ccctatttgt ttatttttct
     2461 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     2521 attgaaaaag gagcttatca ccgataagga ggttttttaa tgacggtgct gcactcggtt
     2581 gacttcttcc ctagcggcaa tgccagcgtt gccattgagc cgcgcctgcc tcaagccgac
     2641 ttcccggagc accaccacga cttccacgag atcgttatcg tggagcacgg taccggcatc
     2701 cacgttttca acggccaacc gtacacgatt acgggcggta ccgtgtgctt cgttcgtgat
     2761 cacgaccgcc acttatacga gcacacggac aacttatgct taaccaacgt tttataccgt
     2821 agccctgacc gcttccaatt cctggcgggc ttaaaccaac tgttaccgca ggaattagac
     2881 ggccaatacc ctagccattg gcgtgtgaat cattcggtgc tgcaacaagt tcgccaatta
     2941 gtggcgcaaa tggagcaaca agagggcgag aacgacctgc cgagcacggc gagccgtgaa
     3001 attctgttca tgcagctgtt actgctgtta cgcaagtcga gcctgcaaga aaatttagag
     3061 aattcggcga gccgcctgaa tctgctgtta gcgtggttag aagatcactt cgcggacgaa
     3121 gttaactggg atgcggttgc cgaccagttc agcctgagct tacgcaccct gcaccgccaa
     3181 ctgaaacaac agaccggctt aaccccgcaa cgctatttaa accgtttacg cttaatgaag
     3241 gcgcgccact tactgcgcca ttcggaagcg tcggtgaccg atattgcgta ccactgtggc
     3301 ttttcggata gcaatcattt cagcaccctg ttccgtcgcg aattcaattg gagccctcgc
     3361 gacatccgcc aaggccgcga cggtttctta cagtgaaggc cttctagagt cgacctgcat
     3421 cgacctgcag ttgacaatta atcatcggct cgtataatgt gtggaaaggc ggttcaccgc
     3481 cgttttcaca caggaaacag aattctttaa gaaggagata tacatatgtc agtgatagca
     3541 aagcagatga catacaaagt atatatgagc ggtactgtaa atggtcacta cttcgaagta
     3601 gaaggtgatg gcaaagggaa gccgtatgag ggtgaacaaa cagtgaagct tacagttacg
     3661 aagggtggcc ctttgccgtt cgcgtgggac attttgtcgc cacagtgcca gtacgggagc
     3721 ataccattca ccaagtatcc agaggacata ccagactacg tgaaacagtc ctttcccgaa
     3781 ggctatacct gggagcgcat aatgaacttt gaagacggtg cggtttgtac ggtatcgaac
     3841 gattcatcaa tccaaggaaa ttgctttatt tatcatgtta aattttcggg gcttaacttt
     3901 ccgccaaatg gccccgtgat gcagaagaaa actcagggct gggaaccaaa taccgagcgc
     3961 cttttcgctc gggacggaat gctgttggga aacaatttta tggcgttgaa gctggaaggg
     4021 ggtggccact atctctgtga attcaagaca acatacaaag ccaagaagcc tgtcaagatg
     4081 cccggttatc actatgtgga ccggaagctc gatgtcacta atcacaacaa ggattatact
     4141 tcagttgaac agtgcgaaat ttcgatcgct cgcaaacctg tcgtagcata aggggcatgc
     4201 aagcttggct gttttggcgg atgagagggt accgcatgag cttcgagcta atcgcgcaaa
     4261 aaaccccgct tcggcggggt tttttcgttg atcacgtacg atatcttcga accggtgcac
     4321 atggtgtaca gggccctagg ataggacgtc ttaaggttta aaccaggtta attaatttaa
     4381 atgcatcccg ggacgtctcg agctcgatcg gaccgcggcc gctagcacgt ataccaagtg
     4441 tcctgttttc ttgccgccaa ggatctgatg gcgcagggga tcaagatctc tgttgatacc
     4501 gggaagccct gggccaactt ttggcgaaaa tgagacgttg atcggcacgt aagaggttcc
     4561 aactttcacc ataatgaaat aagatcacta ccgggcgtat tttttgagtt atcgagattt
     4621 tcaggagcta aggaagctaa aatggagaaa aaaatcactg gatataccac cgttgatata
     4681 tcccaatggc atcgtaaaga acattttgag gcatttcagt cagttgctca atgtacctat
     4741 aaccagaccg ttcagctgga tattacggcc tttttaaaga ccgtaaagaa aaataagcac
     4801 aagttttatc cggcctttat tcacattctt gcccgcctga tgaatgctca tccggaatta
     4861 cgtatggcaa tgaaagacgg tgagctggtg atatgggata gtgttcaccc ttgttacacc
     4921 gttttccatg agcaaactga aacgttttca tcgctctgga gtgaatacca cgacgatttc
     4981 cggcagtttc tacacatata ttcgcaagat gtggcgtgtt acggtgaaaa cctggcctat
     5041 ttccctaaag ggtttattga gaatatgttt ttcgtctcag ccaatccctg ggtgagtttc
     5101 accagttttg atttaaacgt ggccaatatg gacaacttct tcgcccccgt tttcaccatg
     5161 ggcaaatatt atacgcaagg cgacaaggtg ctgatgccgc tggcgattca ggttcatcat
     5221 gccgtttgtg atggcttcca tgtcggcaga atgcttaatg aattacaaca gtactgcgat
     5281 gagtggcagg gcggggcgta atttttttaa ggcagttatt gtcttcaaat tcccgttgca
     5341 tcgatggccc ccgatggtag tgtggggtct ccccatgcga gagtagggaa ctgccaggca
     5401 tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc
     5461 ggtgaacgct ctcctgagta ggacaaatcc gccgggagcg gatttgaacg ttgcgaagca
     5521 acggcccgga gggtggcggg caggacgccc gccataaact gccaggcatc aaattaagca
     5581 gaaggccatc ctgacggatg gcctttttgc gtggccagtg ccaagcttgc atgcctgcag
     5641 ctgtccggaa ctagtcgagg ccatggtggc ctcctgatgt cgtcaacacg gcgaaatagt
     5701 aatcacgagg tcaggttctt accttaaatt ttcgacggaa aaccacgtaa aaaacgtcga
     5761 tttttcaaga tacagcgtga attttcagga aatgcggtga gcatcacatc accacaattc
     5821 agcaaattgt gaacatcatc acgttcatct ttccctggtt gccaatggcc cattttcctg
     5881 tcagtaacga gaaggtcgcg aattcaggcg ctttttagac tggtcgtaat gaaattcagc
     5941 aggatcacca tatggaaaaa cgcacaccac atacacgttt gagtcaggtt aaaaaacttg
     6001 tcaatgccgg gcaagttcgt acaacacgta gtgccctgtt aaatgcagat gagttaggtt
     6061 tggattttga tggtatgtgt aatgttatca ttggattatc agagagcgac ttttataaaa
     6121 gcatgaccac ctactctgat catactatct ggcaggatgt ttacagaccc aggcttgtta
     6181 caggccaggt ttatcttaaa attacggtaa ttcatgacgt actgatcgtc tcgtttaagg
     6241 agaagtaagg atccgcgaat tctcatgttt gacagcttat cactgatcag tgaattaatg
     6301 gcgatgacgc atcctcacga taatatccgg gtaggcgcaa tcactttcgt ctctactccg
     6361 ttacaaagcg aggctgggta tttcccggcc tttctgttat ccgaaatcca ctgaaagcac
     6421 agcggctggc tgaggagata aataataaac gaggggctgt atgcacaaag catcttctgt
     6481 tgagttaaga acgagtatcg agatggcaca tagccttgct caaattggaa tcaggtttgt
     6541 gccaatacca gtagaaacag acgaagaagt cgtgggggaa ttccacaaat tgttatccgc
     6601 tcacaattcc acatgtggaa ttccacatgt ggaattccca
//
