LOCUS       sequence-432231-        6856 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Empty backbone for Tn7-site insertion with XylS-Pm expression 
            system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6856)
  AUTHORS   Intorcia V, Sava RL, Schroeder GP, Gebhardt MJ
  TITLE     A series of vectors for inducible gene expression in 
            multidrug-resistant Acinetobacter baumannii.
  JOURNAL   Appl Environ Microbiol. 2024 Sep 18;90(9):e0047424. doi: 
            10.1128/aem.00474-24. Epub 2024 Aug 20.
  PUBMED    39162403
REFERENCE   2  (bases 1 to 6856)
  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."; date: "2024-09-18"; volume: "90(9):e0047424. 
            doi"; pages: " 10.1128/aem.00474-24. Epub 2024 Aug 20"
COMMENT     Sequence Label: pUC18T-mini-Tn7T-hph-Ptol
FEATURES             Location/Qualifiers
     source          1..6856
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    221..268
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             complement(332..1297)
                     /codon_start=1
                     /gene="xylS"
                     /product="XylS regulator encoded by the Pseudomonas putida 
                     TOL plasmid pWWO"
                     /label=XylS
                     /note="stimulates transcription from the Pm promoter in the
                     presence of aromatic compounds such as m-toluate or 
                     benzoate"
                     /translation="MDFCLLNEKSQIFVHAEPYAVSDYVNQYVGTHSIRLPKGGRPAGR
                     LHHRIFGCLDLCRISYGGSVRVISPGLETCYHLQIILKGHCLWRGHGQEHYFAPGELLL
                     LNPDDQADLTYSEDCEKFIVKLPSVVLDRACSDNNWHKPREGIRFAARHNLQQLDGFIN
                     LLGLVCDEAEHTKSMPRVQEHYAGIIASKLLEMLGSNVSREIFSKGNPSFERVVQFIEE
                     NLKRNISLERLAELAMMSPRSLYNLFEKHAGTTPKNYIRNRKLESIRACLNDPSANVRS
                     ITEIALDYGFLHLGRFAENYRSAFGELPSDTLRQCKKEVA"
     mobile_element  2328..2493
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     oriT            2652..2760
                     /note="incP origin of transfer"
     rep_origin      complement(2992..3580)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3072..3091)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3751..4542,4543..4611))
                     /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     4374..4393
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4612..4716)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4784..4802
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4840..4862)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4962..4981
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5395..5411
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     5396..5412
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      5440..5534
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      5637..5723
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     protein_bind    5769..5816
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
ORIGIN
        1 ccggcttaat tccgcacggc cggtcgcgac acggcctgtc cgcaccgcgg atcaggcgtt
       61 gacgatgacg ggctggtcgg ccacgtcggg gagtcaggca actatggatg aacgaaatag
      121 acagatcgct gatcagcggc ccggaaacgt cctcgaaacg acgcatgtgt tcctcctggt
      181 tggtacaggt ggttgggggt gctcggctgt cgctggtgtt gaagttccta ttccgaagtt
      241 cctattctct agaaagtata ggaacttcag agcgcttttg aagctaattc gatcatgcat
      301 gagctcgaat tcccggggat cccgttcgta atcaagccac ttcctttttg cattgacgca
      361 gggtgtcgga aggcaactcg ccgaacgcgc tcctatagtt ttcagcgaag cgtcccaaat
      421 gtaagaagcc gtagtctagg gctatctcag ttatactacg cacattggca ctgggatcgt
      481 tcaagcaggc gcggatgctt tcgagcttgc ggttgcggat gtagttcttc ggcgtggtgc
      541 cggcatgctt ctcgaacaaa ttgtagagcg agcgtggact catcatcgcc agctccgcta
      601 accgctcaag gctgatattc cgtttgagat tctcctcaat gaattgaacg actcgctcga
      661 aagacgggtt acctttgctg aaaatttcac ggctgacatt gctgcccagc atttcgagca
      721 gcttggaagc gatgatcccc gcatagtgct cttggacccg aggcatcgac tttgtatgtt
      781 ccgcttcgtc acaaactaac ccgagtagat tgataaagcc atcgagttgc tggagattgt
      841 gtcgcgcggc gaaacggata ccctccctcg gcttgtgcca attgttgtca ctgcatgccc
      901 gatcaaggac cactgagggc aatttaacga taaatttctc gcaatcttct gaataggtca
      961 ggtcggcttg gtcatccgga ttgagcagca atagttcgcc cggcgcaaaa tagtgctcct
     1021 ggccatggcc acgccacagg caatggcctt tgagtattat ttgcagatga taacaggtct
     1081 ctaatccagg cgagattacc ctcacgctac cgccgtagct gattcgacac aggtcgaggc
     1141 atccgaagat tctgtggtgc agcctgcctg ccgggcgccc gcccttgggc aggcgaatag
     1201 agtgcgtacc gacatactgg ttaacataat cggagactgc atagggctcg gcgtggacga
     1261 agatctgact tttctcgttc aataagcaaa aatccatagt tcacggttct cttattttaa
     1321 tgtgggctgc ttggtgtgat gtagaaaggc gccaagtcga tgaaaatgca tctcgacgtg
     1381 atgcgtatac gggttacccc cattgccacg ttgcgccatc ctttttgcaa tcagtgacca
     1441 cttttccaag caaaaataac gccaagcaga acgaagacgt tctttttaag aagcgagaac
     1501 accagaagtt cgtgctgtcg gggcatgcgg cgacgaattg gcggataaag gggatctgct
     1561 ggatattacg gcctttttaa agaccgtaaa gaaaaataag cacaagtttt atccggcctt
     1621 tattcacatt cttgcccgcc tgatgaatgc tcatccggaa ttacgtatgg caatgaaaga
     1681 cggtgagctg gtgatatggg atagtgttca cccttgttac accgttttcc atgagcaaac
     1741 tgaaacgttt tcatcgctct ggagtgaata ccacgacgat ttccggcagt ttctacacat
     1801 atattcgcaa gatgtggcgt gttacggtga aaacctggcc tatttcccta aagggtttat
     1861 tgagaatatg tttttcgtct cagccaatcc ctgggtgagt ttcaccagtt ttgatttaaa
     1921 cgtggccaat atggacaact tcttcgcccc cgttttcacc atgcatggga attagcttga
     1981 tctgaccaac gaccggtagc ggagctatcc aacggcggta taccaggaaa acacacagca
     2041 ggtacatcag aacagtacca tgactgaaga acaaatagtt ttttcctgat ccataaagca
     2101 gaacggcctg ctccatgaca aatctggctc cccaactaat gccccatgca gccagcataa
     2161 ccagcataaa ctgtccggtt tgatagggat aagtccagcc ttgcaagaag cggatacagg
     2221 agtgcaaaaa atggctatct ctagaaaggc ctacccctta ggctggatcc actgcaggaa
     2281 ttcctcgaga agcttgggcc cggtacctcg cgaaggcctt gcaggccaac cagataagtg
     2341 aaatctagtt ccaaactatt ttgtcatttt taattttcgt attagcttac gacgctacac
     2401 ccagttccca tctattttgt cactcttccc taaataatcc ttaaaaactc catttccacc
     2461 cctcccagtt cccaactatt ttgtccgccc acagcggggc atttttcttc ctgttatgtt
     2521 tgggcgctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     2581 tcttccgctt cctcgctcac tgacccgctg cgctcggtcg ttcggctgcg gcgagcggta
     2641 tcagagctta tcggccagcc tcgcagagca ggattcccgt tgagcaccgc caggtgcgaa
     2701 taagggacag tgaagaagga acacccgctc gcgggtgggc ctacttcacc tatcctgccc
     2761 ggctgacgcc gttggataca ccaaggaaag tctacacgaa ccctttggca aaatcctgta
     2821 tatcgtgcga aaaaggatgg atataccgaa aaaatcgcta taatgacccc gaagcagggt
     2881 tatgcagcgg aaagtatacc ttaaggaatc ccctgataac gcaggaaaga acatgtgagc
     2941 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     3001 gctccgcccc cctgacgagc atcacaaaaa tcgacgctcg agtcagaggt ggcgaaaccc
     3061 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     3121 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     3181 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     3241 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     3301 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     3361 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     3421 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     3481 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     3541 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     3601 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     3661 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     3721 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     3781 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     3841 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     3901 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     3961 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     4021 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     4081 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     4141 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     4201 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     4261 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     4321 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     4381 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     4441 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     4501 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     4561 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     4621 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     4681 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     4741 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     4801 gccctttcgt ctcgcgcgtt tcggtgacga cggtgaaaac ctctgacaca tgcagctccc
     4861 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
     4921 gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt
     4981 actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
     5041 catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
     5101 ctcttcgcta ttacgccagc tagaggacca gccgcgtaac ctggcaaaat cggttacggt
     5161 tgagtaataa atggatgccc tgcgtaagca ggtgtgggcg gacaataaag tcttaaactg
     5221 aacaaaatag atctaaacta tgacaataaa gtcttaaact agacagaata gttgtaaact
     5281 gaaatcagtc cagttatgct gtgaaaaagc atactggact tttgttatgg ctaaagcaaa
     5341 ctcttcattt tctgaagtgc aaattgcccg tcgtattaaa gaggggcgtg gggttcgagg
     5401 tcgacggtat cgataagcta gcttaattag ctgagcttgg actcctgttg atagatccag
     5461 taatgacctc agaactccat ctggatttgt tcagaacgct cggttgccgc cgggcgtttt
     5521 ttattggtga gaatccaagc tagactgcga tgagtggcag ggcggggcgt aattttttta
     5581 aggcagttat tggtgccctt aaacgcctgg ggtaatgact ctctagcttg aggcatcaaa
     5641 taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga
     5701 acgctctcct gagtaggaca aatccgccgc taggagcttg cggcccggac gatcgaattg
     5761 gggatcttga agtacctatt ccgaagttcc tattctctag aaagtatagg aacttcagct
     5821 agagggggcg tcaggcgccg ggggcggtgt ccggcggccc ccagaggaac tgcgccagtt
     5881 cctccggatc ggtgaagccg gagagatcca gcggggtctc ctcgaacacc tcgaagtcgt
     5941 gcaggaaggt gaaggcgagc agttcgcggg cgaagtcctc ggtccgcttc cactgcgccc
     6001 cgtcgagcag cgcggccagg atctcgcggt cgccccggaa ggcgttgaga tgcagttgca
     6061 ccaggctgta gcgggagtct cccgcataga cgtcggtgaa gtcgacgatc ccggtgacct
     6121 cggtcgcggc caggtccacg aagatgttgg tcccgtgcag gtcgccgtgg acgaaccggg
     6181 gttcgcggcc ggccagcagc gtgtccacgt ccggcagcca gtcctccagg cggtccagca
     6241 gccggggcga gaggtagccc cacccgcggt ggtcctcgac ggtcgccgcg cggcgttccc
     6301 gcagcagttc cgggaagacc tcggaatggg gggtgagcac ggtgttcccg gtcagcggca
     6361 ccctgtgcag ccggccgagc acccggccga gttcgcgggc cagggcgagc agcgcgttcc
     6421 ggtcggtcgt gccgtccatc gcggaccgcc aggtggtgcc ggtcatccgg ctcatcacca
     6481 ggtagggcca cggccaggct ccggtgccgg gccgcagctc gccgcggccg aggaggcggg
     6541 gcaccggcac cggggcgtcc gccaggaccg cgtacgcctc cgactccgac gcgaggctct
     6601 ccggaccgca ccagtgctcg ccgaacagct tgatcaccgg gccgggctcg ccgaccagta
     6661 cggggttggt gctctcgccg ggcacccgca gcaccggcgg caccggcagc ccgagctcct
     6721 ccagggctcg gcgggccagc ggctcccaga attcctggtc gttccgcagg ctcgcgtagg
     6781 aatcatccga atcaatacgg tcgagaagta acagggattc ttgtgtcaca gcggacctct
     6841 attcacaggg tacggg
//
