LOCUS       sequence-181626-        4030 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Broad host range allelic exchange suicide vector for bacteria - Tse2
            toxin driven by anhydrotetracycline inducible promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4030)
  AUTHORS   Armbruster CE, Forsyth-DeOrnellas V, Johnson AO, Smith SN, Zhao L, 
            Wu W, Mobley HLT
  TITLE     Genome-wide transposon mutagenesis of Proteus mirabilis: Essential 
            genes, fitness factors for catheter-associated urinary tract 
            infection, and the impact of polymicrobial infection on fitness 
            requirements.
  JOURNAL   PLoS Pathog. 2017 Jun 14;13(6):e1006434. doi: 
            10.1371/journal.ppat.1006434.
  PUBMED    28614382
REFERENCE   2  (bases 1 to 4030)
  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: "PLoS 
            Pathog."; date: "2017-06-14"; volume: "13(6):e1006434. doi"; pages: 
            " 10.1371/journal.ppat.1006434"
COMMENT     Sequence Label: pAOJ15
FEATURES             Location/Qualifiers
     source          1..4030
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(322..1113,1114..1182))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     945..964
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1183..1287)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2020..2042
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2034..2051
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2035..2051
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    2052..2108
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(2121..2137)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2121..2137)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2134..2156)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2145..2161
                     /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)."
     promoter        complement(join(2169..2175,2176..2193,2194..2199))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2214..2235
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     oriT            2684..2793
                     /note="incP origin of transfer"
     CDS             complement(2953..3603)
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline resistance regulatory protein"
                     /label=TetR
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKGDD"
ORIGIN
        1 agctacaaca agcgcctgca agccacccac tacaccatca tgccggccaa gccgatgtac
       61 cgggaggtcc tcatgggcca actggacaac ttcgtgcgca acgccatccg ccgccaatgg
      121 gaaaaagccc gcgggctcta ggagtgaacc atgaacctga aaccccagac cgaaccatga
      181 gaccgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
      241 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
      301 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
      361 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
      421 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgcgacccac gctcaccggc
      481 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
      541 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
      601 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
      661 gtcgtttggt atggcttcat tcagctccgg ttcccagcga tcaaggcgag ttacatgatc
      721 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
      781 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
      841 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
      901 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
      961 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     1021 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     1081 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     1141 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     1201 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     1261 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctaaattgta
     1321 agcggtctcc tgccattttt ggggtgaggc cgttcgcggc cgaggggcgc agcccctggg
     1381 gggatgggag gcccgcgtta gcgggccggg agggttcgag aagggggggc accccccttc
     1441 ggcgtgcgcg gtcacgcgca cagggcgcag ccctggttaa aaacaaggtt tataaatatt
     1501 ggtttaaaag caggttaaaa gacaggttag cggtggccga aaaacgggcg gaaacccttg
     1561 caaatgctgg attttctgcc tgtggacagc ccctcaaatg tcaataggtg cgcccctcat
     1621 ctgtcagcac tctgcccctc aagtgtcaag gatcgcgccc ctcatctgtc agtagtcgcg
     1681 cccctcaagt gtcaataccg cagggcactt atccccaggc ttgtccacat catctgtggg
     1741 aaactcgcgt aaaatcaggc gttttcgccg atttgcgagg ctggccagct ccacgtcgcc
     1801 ggccgaaatc gagcctgccc ctcatctgtc aacgccgcgc cgggtgagtc ggcccctcaa
     1861 gtgtcaacgt ccgcccctca tctgtcagtg agggccaagt tttccgcgag gtatccaact
     1921 gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat
     1981 gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa
     2041 cgacggccag tgaattcgag ctcggtaccc ggggatcctc tagagtcgac ctgcaggcat
     2101 gcaagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     2161 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     2221 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     2281 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     2341 tcggtcttgc cttgctcgtc ggtgatgtac ttcaccagct ccgcgaagtc gctcttcttg
     2401 agtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     2461 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     2521 agctaccaac tcttaacccg gcctcagcgc cgggttttct ttgcctcacg atcggctctt
     2581 cgttcgtctg gaaggcagta caccttgata ggtgggctgc ccttcctggt tggcttggtt
     2641 tcatcagcca tccgcttgcc ctcatctgtt acgccggcgg tagccggcca gcctcgcaga
     2701 gcaggattcc cgttgagcac cgccaggtgc gaataaggga cagtgaagaa ggaacacccg
     2761 ctcgcgggtg ggcctacttc acctatcctg cccggctgac gccgttggat acaccaagga
     2821 aagtctacac gaagggcatg aaaaagcccg tagcgggctg ctacgggcgt ctgacgcggt
     2881 ggaaaggggg aggggatgtt gtctacatgg ctctgctgta gtgagtgggt tgcgctccgg
     2941 cagcggtcct gatcaatcgt caccctttct cggtccttca acgttcctga caacgagcct
     3001 ccttttcgcc aatccatcga caatcaccgc gagtccctgc tcgaacgctg cgtccggacc
     3061 ggcttcgtcg aaggcgtcta tcgcggcccg caacagcggc gagagcggag cctgttcaac
     3121 ggtgccgccg cgctcgccgg catcgctgtc gccggcctgc tcctcaagca cggccccaac
     3181 agtgaagtag ctgattgtca tcagcgcatt gacggcgtcc ccggccgaaa aacccgcctc
     3241 gcagaggaag cgaagctgcg cgtcggccgt ttccatctgc ggtgcgcccg gtcgcgtgcc
     3301 ggcatggatg cgcgcgccat cgcggtaggc gagcagcgcc tgcctgaagc tgcgggcatt
     3361 cccgatcaga aatgagcgcc agtcgtcgtc ggctctcggc accgaatgcg tatgattctc
     3421 cgccagcatg gcttcggcca gtgcgtcgag cagcgcccgc ttgttcctga agtgccagta
     3481 aagcgccggc tgctgaaccc ccaaccgttc cgccagtttg cgtgtcgtca gaccgtctac
     3541 gccgacctcg ttcaacaggt ccagggcggc acggatcact gtattcggct gcaactttgt
     3601 catgcttgac actttatcac tgataaacat aatatgtcca ccaacttatc agtgataaag
     3661 aatccgcgcg ttcaatccac agggaagtcc acgcatgtcc tacgactacg agaaaaccag
     3721 cctcaccctc taccgggcgg tattcaaggc caactacgac ggcgacgtcg gtcgctacct
     3781 gcgccccgac caggaactcg ccgaggctgc ggaagtcgcc ccgctgctgc atccgacctt
     3841 cgacagcccc aacacccctg gcgtccccgc ccgcgcgccg gacatcgtcg ccggccgcga
     3901 cggcctctac gccccggaca ccggcggcac ctcggtgttc gaccgcgccg gcgtgctgcg
     3961 ccgcgccgac ggcgacttcg tgatacccga cggcaccgac atcccgccgg accttaaggt
     4021 gaaacaggac
//
