LOCUS       sequence-251147-        5878 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes Tse2 toxin. Gent-R.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5878)
  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 5878)
  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: pTOX12
FEATURES             Location/Qualifiers
     source          1..5878
                     /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      696..942
                     /label=lambda tL3 terminator
                     /note="transcription terminator tL3 from phage lambda"
     protein_bind    978..994
                     /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      1029..1417
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     CDS             complement(2035..2406)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            2439..2548
                     /note="incP origin of transfer"
     primer_bind     complement(3229..3248)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(3387..3409)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     RBS             3573..3581
                     /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)"
     CDS             complement(4721..5254)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     terminator      5471..5557
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5649..5676
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     regulatory      5730..5739
                     /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 atatgaggag gtaaaataga tgtcctacga ctacgagaaa
      181 accagcctca ccctctaccg ggcggtattc aaggccaact acgacggcga cgtcggtcgc
      241 tacctgcatc ccgacaagga actcgccgag gctgcggaag tcgccccgct gctgcatccg
      301 accttcgaca gccccaacac ccctggcgtc cccgcccgcg cgccggacat cgtcgccggc
      361 cgcgacggcc tctacgcccc ggacaccggc ggcacctcgg tgttcgaccg cgccggcgtg
      421 ctgcgccgcg ccgacggcga cttcgtgata cccgacggca ccgacatccc gccggacctt
      481 aaggtgaagc aggacagcta caacaagcgc ctgcaagcca cccactacac catcatgccg
      541 gccaagccga tgtaccggga ggtcctcatg ggccaactgg acaacttcgt gcgcaacgcc
      601 atccgccgcc aatgggaaaa agcccgcggg ctctagggat ccgcgaattc tcatgtttga
      661 cagcttatca ctgatcagtg aattaatggc gatgacgcat cctcacgata atatccgggt
      721 aggcgcaatc actttcgtct ctactccgtt acaaagcgag gctgggtatt tcccggcctt
      781 tctgttatcc gaaatccact gaaagcacag cggctggctg aggagataaa taataaacga
      841 ggggctgtat gcacaaagca tcttctgttg agttaagaac gagtatcgag atggcacata
      901 gccttgctca aattggaatc aggtttgtgc caataccagt agaaacagac gaagaagtcg
      961 tgggggaatt ccacaaattg ttatccgctc acaattccac atgtggaatt ccacatgtgg
     1021 aattcccatg tcagccgtta agtgttcctg tgtcactcaa aattgctttg agaggctcta
     1081 agggcttctc agtgcgttac atccctggct tgttgtccac aaccgttaaa ccttaaaagc
     1141 tttaaaagcc ttatatattc ttttttttct tataaaactt aaaaccttag aggctattta
     1201 agttgctgat ttatattaat tttattgttc aaacatgaga gcttagtacg tgaaacatga
     1261 gagcttagta cgttagccat gagagcttag tacgttagcc atgagggttt agttcgttaa
     1321 acatgagagc ttagtacgtt aaacatgaga gcttagtacg tgaaacatga gagcttagta
     1381 cgtactatca acaggttgaa ctgctgatct tcagatcctc tacgccggac gcatcgtggc
     1441 cggatccagc cgaccaggct ttccacgccc gcgtgccgct ccatgtcgtt cgcgcggttc
     1501 tcggaaacgc gctgccgcgt ttcgtgattg tcacgctcaa gcccgtagtc ccgttcgagc
     1561 gtcgcgcaga ggtcagcgag ggcgcggtag gcccgatacg gctcatggat ggtgtttcgg
     1621 gtcgggtgaa tcttgttgat ggcgatatgg atgtgcaggt tgtcggtgtc gtgatgcacg
     1681 gcactgacgc gctgatgctc ggcgaagcca agcccagcgc agatgcggtc ctcaatcgcg
     1741 cgcaacgtct ccgcgtcggg cttctctccc gcgcggaagc taaccagcag gtgataggtc
     1801 ttgtcggcct cggaacgggt gttgccgtgc tgggtcgcca tcacctcggc catgacagcg
     1861 ggcagggtgt ttgcctcgca gttcgtgacg cgcacgtaac ccaggcgctc ggtcttgcct
     1921 tgctcgtcgg tgatgtactt caccagctcc gcgaagtcgc tcttcttgat ggagcgcatg
     1981 gggacgtgct tggcaatcac gcgcaccccc cggccgtttt agcggctaaa aaagtcatgg
     2041 ctctgccctc gggcggacca cgcccatcat gaccttgcca agctcgtcct gcttctcttc
     2101 gatcttcgcc agcagggcga ggatcgtggc atcaccgaac cgcgccgtgc gcgggtcgtc
     2161 ggtgagccag agtttcagca ggccgcccag gcggcccagg tcgccattga tgcgggccag
     2221 ctcgcggacg tgctcatagt ccacgacgcc cgtgattttg tagccctggc cgacggccag
     2281 caggtaggcc gacaggctca tgccggccgc cgccgccttt tcctcaatcg ctcttcgttc
     2341 gtctggaagg cagtacacct tgataggtgg gctgcccttc ctggttggct tggtttcatc
     2401 agccatccgc ttgccctcat ctgttacgcc ggcggtagcc ggccagcctc gcagagcagg
     2461 attcccgttg agcaccgcca ggtgcgaata agggacagtg aagaaggaac acccgctcgc
     2521 gggtgggcct acttcaccta tcctgcccgg ctgacgccgt tggatacacc aaggaaagtc
     2581 tacacgaacc ctttggcaaa atcctgtata tcgtgcgaaa aaggatggat ataccgaaaa
     2641 aatcgctata atgaccccga agcagggtta tgcagcggaa aagcgctgct tccctgctgt
     2701 tttgtggaat atctaccgac tggaaacagg caaatgcagg aaattactga actgagggga
     2761 caggcgagag acgatgccaa agagctacac cgacgagctg gccgagtggg ttgaatcccg
     2821 cgcggccaag aagcgccggc gtgatgaggc tgcggttgcg ttcctggcgg tgagggcgga
     2881 tgtcgaggcg gcgttagcgt ccggctatgc gctcgtcacc atttgggagc acatgcggga
     2941 aacggggaag gtcaagttct cctacgagac gttccgctcg cacgccaggc ggcacatcaa
     3001 ggccaagccc gccgatgtgc ccgcaccgca ggccaaggct gcggaacccg cgccggcacc
     3061 caagacgccg gagccacggc ggccgaagca ggggggcaag gctgaaaagc cggcccccgc
     3121 tgcggccccg accggcttca ccttcaaccc aacaccggac aaaaaggatc ctctacgccg
     3181 gacgcatcgt ggccggcatc accggcgcca caggtgcggt tgctggcgcc tatatcgccg
     3241 acatcaccga tggggaagat cgggctcgcc acttcgggct catgagcgct tgtttcggcg
     3301 tgggtatggt ggcaggcccc gtggccgggg gactgttggg cgccatctcc ttgctgcctc
     3361 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
     3421 gcttgtctgt aagcggatgc cgtaagtaag gtaaaggcct cgcggaaccc ctatttgttt
     3481 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     3541 tcaataatat tgaaaaagga gcttatcacc gataaggagg ttttttaatg acggtgctgc
     3601 actcggttga cttcttccct agcggcaatg ccagcgttgc cattgagccg cgcctgcctc
     3661 aagccgactt cccggagcac caccacgact tccacgagat cgttatcgtg gagcacggta
     3721 ccggcatcca cgttttcaac ggccaaccgt acacgattac gggcggtacc gtgtgcttcg
     3781 ttcgtgatca cgaccgccac ttatacgagc acacggacaa cttatgctta accaacgttt
     3841 tataccgtag ccctgaccgc ttccaattcc tggcgggctt aaaccaactg ttaccgcagg
     3901 aattagacgg ccaataccct agccattggc gtgtgaatca ttcggtgctg caacaagttc
     3961 gccaattagt ggcgcaaatg gagcaacaag agggcgagaa cgacctgccg agcacggcga
     4021 gccgtgaaat tctgttcatg cagctgttac tgctgttacg caagtcgagc ctgcaagaaa
     4081 atttagagaa ttcggcgagc cgcctgaatc tgctgttagc gtggttagaa gatcacttcg
     4141 cggacgaagt taactgggat gcggttgccg accagttcag cctgagctta cgcaccctgc
     4201 accgccaact gaaacaacag accggcttaa ccccgcaacg ctatttaaac cgtttacgct
     4261 taatgaaggc gcgccactta ctgcgccatt cggaagcgtc ggtgaccgat attgcgtacc
     4321 actgtggctt ttcggatagc aatcatttca gcaccctgtt ccgtcgcgaa ttcaattgga
     4381 gccctcgcga catccgccaa ggccgcgacg gtttcttaca gtgaaggcct tctagagtcg
     4441 acctgcaggc atgcacgctt ggctgttttg gcggatgaga gggtaccgca tgagcttcga
     4501 gctaatcgcg caaaaaaccc cgcttcggcg gggttttttc gctgatcacg tacgatatct
     4561 tcgaaccggt gcacatggtg tacagggccc taggatagga cgtcttaagg tttaaaccag
     4621 gttaattaat ttaaatgcat cccgggacgt ctcgagctcg atcggaccgc ggccgctagc
     4681 acgtatacca agtgtcctgt tttcttgccg ccaaggatct ttaggtggcg gtacttgggt
     4741 cgatatcaaa gtgcatcact tcttcccgta tgcccaactt tgtatagaga gccactgcgg
     4801 gatcgtcacc gtaatctgct tgcacgtaga tcacataagc accaagcgcg ttggcctcat
     4861 gcttgaggag attgatgagc gcggtggcaa tgccctgcct ccggtgctcg ccggagactg
     4921 cgagatcata gatatagatc tcactacgcg gctgctcaaa cttgggcaga acgtaagccg
     4981 cgagagcgcc aacaaccgct tcttggtcga aggcagcaag cgcgatgaat gtcttactac
     5041 ggagcaagtt cccgaggtaa tcggagtccg gctgatgttg ggagtaggtg gctacgtctc
     5101 cgaactcacg accgaaaaga tcaagagcag cccgcatgga tttgacttgg tcagggccga
     5161 gcctacatgt gcgaatgatg cccatacttg agccacctaa ctttgtttta gggcgactgc
     5221 cctgctgcgt aacatcgttg ctgctgcgta acatatgttt ttcctcctta tgttaagctt
     5281 actcagttat tatatcataa atatctgtgt caagaataaa ctcccacatg gattcgacag
     5341 tactgcgatg agtggcaggg cggggcgtaa tttttttaag gcagttattg tcttcaaatt
     5401 cccgttgcat cgatggcccc cgatggtagt gtggggtctc cccatgcgag agtagggaac
     5461 tgccaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg
     5521 ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg ccgggagcgg atttgaacgt
     5581 tgcgaagcaa cggcccggag ggtggcgggc aggacgcccg ccataaactg ccaggcatca
     5641 aattaagcag aaggccatcc tgacggatgg cctttttgcg tggccagtgc caagcttgca
     5701 tgcctgcagc tgtccggaac tagtcgaggc catggtggcc tcctgatgtc gtcaacacgg
     5761 cgaaatagta atcacgaggt caggttctta ccttaaattt tcgacggaaa accacgtaaa
     5821 aaacgtcgat ttttcaatat acagcgtgaa ttttcaggaa atgcggtgag catcacat
//
