LOCUS       sequence-248508-        6034 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes YhaV toxin. CAM-R.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6034)
  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 6034)
  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: pTOX1
FEATURES             Location/Qualifiers
     source          1..6034
                     /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      667..913
                     /label=lambda tL3 terminator
                     /note="transcription terminator tL3 from phage lambda"
     protein_bind    949..965
                     /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      1000..1388
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     CDS             complement(2006..2377)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            2410..2519
                     /note="incP origin of transfer"
     primer_bind     complement(3200..3219)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(3358..3380)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     RBS             3544..3552
                     /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        4764..4866
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             4867..5526
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(4933..4952)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      5627..5713
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5805..5832
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     regulatory      5886..5895
                     /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 atatggattt tccacaaagg gttaatggtt gggcgctata
      181 tgctcatccc tgttttcagg aaacctacga cgctttagtt gccgaagtcg agacattaaa
      241 gggaaaagat cctgaaaatt atcagagaaa agccgccaca aagttattgg cggtagtcca
      301 taaagtgatt gaggagcata tcacggtcaa tccatcatca ccggcattcc gtcatggcaa
      361 gtcgttaggc tctgggaaaa ataaagactg gtcacgggta aaatttggtg ctggtcgtta
      421 tcgtctcttc tttcgttata gtgaaaaaga gaaagtcatc attctgggat ggatgaacga
      481 tgaaaacact ctgcgcacct acggtaaaaa aacagatgcc tataccgtat tcagcaaaat
      541 gttaaaaaga ggacatcctc ctgccgactg ggaaaccctc acccgagaaa cagaagaaac
      601 ccattgagga tccgcgaatt ctcatgtttg acagcttatc actgatcagt gaattaatgg
      661 cgatgacgca tcctcacgat aatatccggg taggcgcaat cactttcgtc tctactccgt
      721 tacaaagcga ggctgggtat ttcccggcct ttctgttatc cgaaatccac tgaaagcaca
      781 gcggctggct gaggagataa ataataaacg aggggctgta tgcacaaagc atcttctgtt
      841 gagttaagaa cgagtatcga gatggcacat agccttgctc aaattggaat caggtttgtg
      901 ccaataccag tagaaacaga cgaagaagtc gtgggggaat tccacaaatt gttatccgct
      961 cacaattcca catgtggaat tccacatgtg gaattcccat gtcagccgtt aagtgttcct
     1021 gtgtcactca aaattgcttt gagaggctct aagggcttct cagtgcgtta catccctggc
     1081 ttgttgtcca caaccgttaa accttaaaag ctttaaaagc cttatatatt cttttttttc
     1141 ttataaaact taaaacctta gaggctattt aagttgctga tttatattaa ttttattgtt
     1201 caaacatgag agcttagtac gtgaaacatg agagcttagt acgttagcca tgagagctta
     1261 gtacgttagc catgagggtt tagttcgtta aacatgagag cttagtacgt taaacatgag
     1321 agcttagtac gtgaaacatg agagcttagt acgtactatc aacaggttga actgctgatc
     1381 ttcagatcct ctacgccgga cgcatcgtgg ccggatccag ccgaccaggc tttccacgcc
     1441 cgcgtgccgc tccatgtcgt tcgcgcggtt ctcggaaacg cgctgccgcg tttcgtgatt
     1501 gtcacgctca agcccgtagt cccgttcgag cgtcgcgcag aggtcagcga gggcgcggta
     1561 ggcccgatac ggctcatgga tggtgtttcg ggtcgggtga atcttgttga tggcgatatg
     1621 gatgtgcagg ttgtcggtgt cgtgatgcac ggcactgacg cgctgatgct cggcgaagcc
     1681 aagcccagcg cagatgcggt cctcaatcgc gcgcaacgtc tccgcgtcgg gcttctctcc
     1741 cgcgcggaag ctaaccagca ggtgataggt cttgtcggcc tcggaacggg tgttgccgtg
     1801 ctgggtcgcc atcacctcgg ccatgacagc gggcagggtg tttgcctcgc agttcgtgac
     1861 gcgcacgtga cccaggcgct cggtcttgcc ttgctcgtcg gtgatgtact tcaccagctc
     1921 cgcgaagtcg ctcttcttga tggagcgcat ggggacgtgc ttggcaatca cgcgcacccc
     1981 ccggccgttt tagcggctaa aaaagtcatg gctctgccct cgggcggacc acgcccatca
     2041 tgaccttgcc aagctcgtcc tgcttctctt cgatcttcgc cagcagggcg aggatcgtgg
     2101 catcaccgaa ccgcgccgtg cgcgggtcgt cggtgagcca gagtttcagc aggccgccca
     2161 ggcggcccag gtcgccattg atgcgggcca gctcgcggac gtgctcatag tccacgacgc
     2221 ccgtgatttt gtagccctgg ccgacggcca gcaggtaggc cgacaggctc atgccggccg
     2281 ccgccgcctt ttcctcaatc gctcttcgtt cgtctggaag gcagtacacc ttgataggtg
     2341 ggctgccctt cctggttggc ttggtttcat cagccatccg cttgccctca tctgttacgc
     2401 cggcggtagc cggccagcct cgcagagcag gattcccgtt gagcaccgcc aggtgcgaat
     2461 aagggacagt gaagaaggaa cacccgctcg cgggtgggcc tacttcacct atcctgcccg
     2521 gctgacgccg ttggatacac caaggaaagt ctacacgaac cctttggcaa aatcctgtat
     2581 atcgtgcgaa aaaggatgga tataccgaaa aaatcgctat aatgaccccg aagcagggtt
     2641 atgcagcgga aaagcgctgc ttccctgctg ttttgtggaa tatctaccga ctggaaacag
     2701 gcaaatgcag gaaattactg aactgagggg acaggcgaga gacgatgcca aagagctaca
     2761 ccgacgagct ggccgagtgg gttgaatccc gcgcggccaa gaagcgccgg cgtgatgagg
     2821 ctgcggttgc gttcctggcg gtgagggcgg atgtcgaggc ggcgttagcg tccggctatg
     2881 cgctcgtcac catttgggag cacatgcggg aaacggggaa ggtcaagttc tcctacgaga
     2941 cgttccgctc gcacgccagg cggcacatca aggccaagcc cgccgatgtg cccgcaccgc
     3001 aggccaaggc tgcggaaccc gcgccggcac ccaagacgcc ggagccacgg cggccgaagc
     3061 aggggggcaa ggctgaaaag ccggcccccg ctgcggcccc gaccggcttc accttcaacc
     3121 caacaccgga caaaaaggat cctctacgcc ggacgcatcg tggccggcat caccggcgcc
     3181 acaggtgcgg ttgctggcgc ctatatcgcc gacatcaccg atggggaaga tcgggctcgc
     3241 cacttcgggc tcatgagcgc ttgtttcggc gtgggtatgg tggcaggccc cgtggccggg
     3301 ggactgttgg gcgccatctc cttgctgcct cgcgcgtttc ggtgatgacg gtgaaaacct
     3361 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgtaagtaa
     3421 ggtaaaggcc tcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     3481 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg agcttatcac
     3541 cgataaggag gttttttaat gacggtgctg cactcggttg acttcttccc tagcggcaat
     3601 gccagcgttg ccattgagcc gcgcctgcct caagccgact tcccggagca ccaccacgac
     3661 ttccacgaga tcgttatcgt ggagcacggt accggcatcc acgttttcaa cggccaaccg
     3721 tacacgatta cgggcggtac cgtgtgcttc gttcgtgatc acgaccgcca cttatacgag
     3781 cacacggaca acttatgctt aaccaacgtt ttataccgta gccctgaccg cttccaattc
     3841 ctggcgggct taaaccaact gttaccgcag gaattagacg gccaataccc tagccattgg
     3901 cgtgtgaatc attcggtgct gcaacaagtt cgccaattag tggcgcaaat ggagcaacaa
     3961 gagggcgaga acgacctgcc gagcacggcg agccgtgaaa ttctgttcat gcagctgtta
     4021 ctgctgttac gcaagtcgag cctgcaagaa aatttagaga attcggcgag ccgcctgaat
     4081 ctgctgttag cgtggttaga agatcacttc gcggacgaag ttaactggga tgcggttgcc
     4141 gaccagttca gcctgagctt acgcaccctg caccgccaac tgaaacaaca gaccggctta
     4201 accccgcaac gctatttaaa ccgtttacgc ttaatgaagg cgcgccactt actgcgccat
     4261 tcggaagcgt cggtgaccga tattgcgtac cactgtggct tttcggatag caatcatttc
     4321 agcaccctgt tccgtcgcga attcaattgg agccctcgcg acatccgcca aggccgcgac
     4381 ggtttcttac agtgaaggcc ttctagagtc gacctgcagg catgcaagct tggctgtttt
     4441 ggcggatgag agggtaccgc atgagcttcg agctaatcgc gcaaaaaacc ccgcttcggc
     4501 ggggtttttt cgctgatcac gtacgatatc ttcgaaccgg tgcacatggt gtacagggcc
     4561 ctaggatagg acgtcttaag gtttaaacca ggttaattaa tttaaatgca tcccgggacg
     4621 tctcgagctc gatcggaccg cggccgctag cacgtatacc aagtgtcctg ttttcttgcc
     4681 gccaaggatc tgatggcgca ggggatcaag atctctgttg ataccgggaa gccctgggcc
     4741 aacttttggc gaaaatgaga cgttgatcgg cacgtaagag gttccaactt tcaccataat
     4801 gaaataagat cactaccggg cgtatttttt gagttatcga gattttcagg agctaaggaa
     4861 gctaaaatgg agaaaaaaat cactggatat accaccgttg atatatccca atggcatcgt
     4921 aaagaacatt ttgaggcatt tcagtcagtt gctcaatgta cctataacca gaccgttcag
     4981 ctggatatta cggccttttt aaagaccgta aagaaaaata agcacaagtt ttatccggcc
     5041 tttattcaca ttcttgcccg cctgatgaat gctcatccgg aattacgtat ggcaatgaaa
     5101 gacggtgagc tggtgatatg ggatagtgtt cacccttgtt acaccgtttt ccatgagcaa
     5161 actgaaacgt tttcatcgct ctggagtgaa taccacgacg atttccggca gtttctacac
     5221 atatattcgc aagatgtggc gtgttacggt gaaaacctgg cctatttccc taaagggttt
     5281 attgagaata tgtttttcgt ctcagccaat ccctgggtga gtttcaccag ttttgattta
     5341 aacgtggcca atatggacaa cttcttcgcc cccgttttca ccatgggcaa atattatacg
     5401 caaggcgaca aggtgctgat gccgctggcg attcaggttc atcatgccgt ttgtgatggc
     5461 ttccatgtcg gcagaatgct taatgaatta caacagtact gcgatgagtg gcagggcggg
     5521 gcgtaatttt tttaaggcag ttattgtctt caaattcccg ttgcatcgat ggcccccgat
     5581 ggtagtgtgg ggtctcccca tgcgagagta gggaactgcc aggcatcaaa taaaacgaaa
     5641 ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcct
     5701 gagtaggaca aatccgccgg gagcggattt gaacgttgcg aagcaacggc ccggagggtg
     5761 gcgggcagga cgcccgccat aaactgccag gcatcaaatt aagcagaagg ccatcctgac
     5821 ggatggcctt tttgcgtggc cagtgccaag cttgcatgcc tgcagctgtc cggaactagt
     5881 cgaggccatg gtggcctcct gatgtcgtca acacggcgaa atagtaatca cgaggtcagg
     5941 ttcttacctt aaattttcga cggaaaacca cgtaaaaaac gtcgattttt caatatacag
     6001 cgtgaatttt caggaaatgc ggtgagcatc acat
//
