LOCUS       sequence-248507-        5866 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes MqsR toxin. CAM-R.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5866)
  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 5866)
  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: pTOX2
FEATURES             Location/Qualifiers
     source          1..5866
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..119
                     /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      499..745
                     /label=lambda tL3 terminator
                     /note="transcription terminator tL3 from phage lambda"
     protein_bind    781..797
                     /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      832..1220
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     CDS             complement(1838..2209)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            2242..2351
                     /note="incP origin of transfer"
     primer_bind     complement(3032..3051)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(3190..3212)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     RBS             3376..3384
                     /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        4596..4698
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             4699..5358
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(4765..4784)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      5459..5545
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5637..5664
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     regulatory      5718..5727
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 caccacaatt cagcaaattg tgaacatcat cacgttcatc tttccctggt tgccaatggc
       61 ccattttcct gtcagtaacg agaaggtcgc gaattcaggc gctttttaga ctggtcgtaa
      121 tgaaattcag caggatcacc atatggaaaa acgcacacca catacacgtt tgagtcaggt
      181 taaaaaactt gtcaatgccg ggcaagttcg tacaacacgt agtgccctgt taaatgcaga
      241 tgagttaggt ttggattttg atggtatgtg taatgttatc attggattat cagagagcga
      301 cttttataaa agcatgacca cctactctga tcatactatc tggcaggatg tttacagacc
      361 caggcttgtt acaggccagg tttatcttaa aattacggta attcatgacg tactgatcgt
      421 ctcgtttaag gagaagtaag gatccgcgaa ttctcatgtt tgacagctta tcactgatca
      481 gtgaattaat ggcgatgacg catcctcacg ataatatccg ggtaggcgca atcactttcg
      541 tctctactcc gttacaaagc gaggctgggt atttcccggc ctttctgtta tccgaaatcc
      601 actgaaagca cagcggctgg ctgaggagat aaataataaa cgaggggctg tatgcacaaa
      661 gcatcttctg ttgagttaag aacgagtatc gagatggcac atagccttgc tcaaattgga
      721 atcaggtttg tgccaatacc agtagaaaca gacgaagaag tcgtggggga attccacaaa
      781 ttgttatccg ctcacaattc cacatgtgga attccacatg tggaattccc atgtcagccg
      841 ttaagtgttc ctgtgtcact caaaattgct ttgagaggct ctaagggctt ctcagtgcgt
      901 tacatccctg gcttgttgtc cacaaccgtt aaaccttaaa agctttaaaa gccttatata
      961 ttcttttttt tcttataaaa cttaaaacct tagaggctat ttaagttgct gatttatatt
     1021 aattttattg ttcaaacatg agagcttagt acgtgaaaca tgagagctta gtacgttagc
     1081 catgagagct tagtacgtta gccatgaggg tttagttcgt taaacatgag agcttagtac
     1141 gttaaacatg agagcttagt acgtgaaaca tgagagctta gtacgtacta tcaacaggtt
     1201 gaactgctga tcttcagatc ctctacgccg gacgcatcgt ggccggatcc agccgaccag
     1261 gctttccacg cccgcgtgcc gctccatgtc gttcgcgcgg ttctcggaaa cgcgctgccg
     1321 cgtttcgtga ttgtcacgct caagcccgta gtcccgttcg agcgtcgcgc agaggtcagc
     1381 gagggcgcgg taggcccgat acggctcatg gatggtgttt cgggtcgggt gaatcttgtt
     1441 gatggcgata tggatgtgca ggttgtcggt gtcgtgatgc acggcactga cgcgctgatg
     1501 ctcggcgaag ccaagcccag cgcagatgcg gtcctcaatc gcgcgcaacg tctccgcgtc
     1561 gggcttctct cccgcgcgga agctaaccag caggtgatag gtcttgtcgg cctcggaacg
     1621 ggtgttgccg tgctgggtcg ccatcacctc ggccatgaca gcgggcaggg tgtttgcctc
     1681 gcagttcgtg acgcgcacgt gacccaggcg ctcggtcttg ccttgctcgt cggtgatgta
     1741 cttcaccagc tccgcgaagt cgctcttctt gatggagcgc atggggacgt gcttggcaat
     1801 cacgcgcacc ccccggccgt tttagcggct aaaaaagtca tggctctgcc ctcgggcgga
     1861 ccacgcccat catgaccttg ccaagctcgt cctgcttctc ttcgatcttc gccagcaggg
     1921 cgaggatcgt ggcatcaccg aaccgcgccg tgcgcgggtc gtcggtgagc cagagtttca
     1981 gcaggccgcc caggcggccc aggtcgccat tgatgcgggc cagctcgcgg acgtgctcat
     2041 agtccacgac gcccgtgatt ttgtagccct ggccgacggc cagcaggtag gccgacaggc
     2101 tcatgccggc cgccgccgcc ttttcctcaa tcgctcttcg ttcgtctgga aggcagtaca
     2161 ccttgatagg tgggctgccc ttcctggttg gcttggtttc atcagccatc cgcttgccct
     2221 catctgttac gccggcggta gccggccagc ctcgcagagc aggattcccg ttgagcaccg
     2281 ccaggtgcga ataagggaca gtgaagaagg aacacccgct cgcgggtggg cctacttcac
     2341 ctatcctgcc cggctgacgc cgttggatac accaaggaaa gtctacacga accctttggc
     2401 aaaatcctgt atatcgtgcg aaaaaggatg gatataccga aaaaatcgct ataatgaccc
     2461 cgaagcaggg ttatgcagcg gaaaagcgct gcttccctgc tgttttgtgg aatatctacc
     2521 gactggaaac aggcaaatgc aggaaattac tgaactgagg ggacaggcga gagacgatgc
     2581 caaagagcta caccgacgag ctggccgagt gggttgaatc ccgcgcggcc aagaagcgcc
     2641 ggcgtgatga ggctgcggtt gcgttcctgg cggtgagggc ggatgtcgag gcggcgttag
     2701 cgtccggcta tgcgctcgtc accatttggg agcacatgcg ggaaacgggg aaggtcaagt
     2761 tctcctacga gacgttccgc tcgcacgcca ggcggcacat caaggccaag cccgccgatg
     2821 tgcccgcacc gcaggccaag gctgcggaac ccgcgccggc acccaagacg ccggagccac
     2881 ggcggccgaa gcaggggggc aaggctgaaa agccggcccc cgctgcggcc ccgaccggct
     2941 tcaccttcaa cccaacaccg gacaaaaagg atcctctacg ccggacgcat cgtggccggc
     3001 atcaccggcg ccacaggtgc ggttgctggc gcctatatcg ccgacatcac cgatggggaa
     3061 gatcgggctc gccacttcgg gctcatgagc gcttgtttcg gcgtgggtat ggtggcaggc
     3121 cccgtggccg ggggactgtt gggcgccatc tccttgctgc ctcgcgcgtt tcggtgatga
     3181 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga
     3241 tgccgtaagt aaggtaaagg cctcgcggaa cccctatttg tttatttttc taaatacatt
     3301 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     3361 ggagcttatc accgataagg aggtttttta atgacggtgc tgcactcggt tgacttcttc
     3421 cctagcggca atgccagcgt tgccattgag ccgcgcctgc ctcaagccga cttcccggag
     3481 caccaccacg acttccacga gatcgttatc gtggagcacg gtaccggcat ccacgttttc
     3541 aacggccaac cgtacacgat tacgggcggt accgtgtgct tcgttcgtga tcacgaccgc
     3601 cacttatacg agcacacgga caacttatgc ttaaccaacg ttttataccg tagccctgac
     3661 cgcttccaat tcctggcggg cttaaaccaa ctgttaccgc aggaattaga cggccaatac
     3721 cctagccatt ggcgtgtgaa tcattcggtg ctgcaacaag ttcgccaatt agtggcgcaa
     3781 atggagcaac aagagggcga gaacgacctg ccgagcacgg cgagccgtga aattctgttc
     3841 atgcagctgt tactgctgtt acgcaagtcg agcctgcaag aaaatttaga gaattcggcg
     3901 agccgcctga atctgctgtt agcgtggtta gaagatcact tcgcggacga agttaactgg
     3961 gatgcggttg ccgaccagtt cagcctgagc ttacgcaccc tgcaccgcca actgaaacaa
     4021 cagaccggct taaccccgca acgctattta aaccgtttac gcttaatgaa ggcgcgccac
     4081 ttactgcgcc attcggaagc gtcggtgacc gatattgcgt accactgtgg cttttcggat
     4141 agcaatcatt tcagcaccct gttccgtcgc gaattcaatt ggagccctcg cgacatccgc
     4201 caaggccgcg acggtttctt acagtgaagg ccttctagag tcgacctgca ggcatgcaag
     4261 cttggctgtt ttggcggatg agagggtacc gcatgagctt cgagctaatc gcgcaaaaaa
     4321 ccccgcttcg gcggggtttt ttcgctgatc acgtacgata tcttcgaacc ggtgcacatg
     4381 gtgtacaggg ccctaggata ggacgtctta aggtttaaac caggttaatt aatttaaatg
     4441 catcccggga cgtctcgagc tcgatcggac cgcggccgct agcacgtata ccaagtgtcc
     4501 tgttttcttg ccgccaagga tctgatggcg caggggatca agatctctgt tgataccggg
     4561 aagccctggg ccaacttttg gcgaaaatga gacgttgatc ggcacgtaag aggttccaac
     4621 tttcaccata atgaaataag atcactaccg ggcgtatttt ttgagttatc gagattttca
     4681 ggagctaagg aagctaaaat ggagaaaaaa atcactggat ataccaccgt tgatatatcc
     4741 caatggcatc gtaaagaaca ttttgaggca tttcagtcag ttgctcaatg tacctataac
     4801 cagaccgttc agctggatat tacggccttt ttaaagaccg taaagaaaaa taagcacaag
     4861 ttttatccgg cctttattca cattcttgcc cgcctgatga atgctcatcc ggaattacgt
     4921 atggcaatga aagacggtga gctggtgata tgggatagtg ttcacccttg ttacaccgtt
     4981 ttccatgagc aaactgaaac gttttcatcg ctctggagtg aataccacga cgatttccgg
     5041 cagtttctac acatatattc gcaagatgtg gcgtgttacg gtgaaaacct ggcctatttc
     5101 cctaaagggt ttattgagaa tatgtttttc gtctcagcca atccctgggt gagtttcacc
     5161 agttttgatt taaacgtggc caatatggac aacttcttcg cccccgtttt caccatgggc
     5221 aaatattata cgcaaggcga caaggtgctg atgccgctgg cgattcaggt tcatcatgcc
     5281 gtttgtgatg gcttccatgt cggcagaatg cttaatgaat tacaacagta ctgcgatgag
     5341 tggcagggcg gggcgtaatt tttttaaggc agttattgtc ttcaaattcc cgttgcatcg
     5401 atggcccccg atggtagtgt ggggtctccc catgcgagag tagggaactg ccaggcatca
     5461 aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt
     5521 gaacgctctc ctgagtagga caaatccgcc gggagcggat ttgaacgttg cgaagcaacg
     5581 gcccggaggg tggcgggcag gacgcccgcc ataaactgcc aggcatcaaa ttaagcagaa
     5641 ggccatcctg acggatggcc tttttgcgtg gccagtgcca agcttgcatg cctgcagctg
     5701 tccggaacta gtcgaggcca tggtggcctc ctgatgtcgt caacacggcg aaatagtaat
     5761 cacgaggtca ggttcttacc ttaaattttc gacggaaaac cacgtaaaaa acgtcgattt
     5821 ttcaagatac agcgtgaatt ttcaggaaat gcggtgagca tcacat
//
