LOCUS       sequence-432234-        5639 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Empty backbone for Tn7-site insertion with tetR-PtetA expression 
            system with two tetR operator sites.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5639)
  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 5639)
  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-Ptet2
FEATURES             Location/Qualifiers
     source          1..5639
                     /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(341..967)
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MMSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWH
                     VKNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGT
                     RPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPT
                     TDSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     872..891
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     promoter        983..1038
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    1019..1037
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     mobile_element  1111..1276
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     oriT            1435..1543
                     /note="incP origin of transfer"
     rep_origin      complement(1775..2363)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1855..1874)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2534..3325,3326..3394))
                     /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     3157..3176
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3395..3499)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3567..3585
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3623..3645)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3745..3764
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4178..4194
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     4179..4195
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      4223..4317
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      4420..4506
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     protein_bind    4552..4599
                     /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 ccatgcatga gctcactagt ttaagaccca ctttcacatt
      361 taagttgttt ttctaatccg catatgatca attcaaggcc gaataagaag gctggctctg
      421 caccttggtg atcaaataat tcgatagctt gtcgtaataa tggcggcata ctatcagtag
      481 taggtgtttc cctttcttct ttagcgactt gatgctcttg atcttccaat acgcaaccta
      541 aagtaaaatg ccccacagcg ctgagtgcat ataatgcatt ctctagtgaa aaaccttgtt
      601 ggcataaaaa ggctaattga ttttcgagag tttcatactg tttttctgta ggccgtgtac
      661 ctaaatgtac ttttgctcca tcgcgatgac ttagtaaagc acatctaaaa cttttagcgt
      721 tattacgtaa aaaatcttgc cagctttccc cttctaaagg gcaaaagtga gtatggtgcc
      781 tatctaacat ctcaatggct aaggcgtcga gcaaagcccg cttatttttt acatgccaat
      841 acaatgtagg ctgctctaca cctagcttct gggcgagttt acgggttgtt aaaccttcga
      901 ttccgacctc attaagcagc tctaatgcgc tgttaatcac tttactttta tctaatctag
      961 acatcattaa ttcctaattt ttgttgacac tctatcgttg atagagttat tttaccactc
     1021 cctatcagtg atagagaaaa gaattcaaaa gatcctgcag gaattcctcg agaagcttgg
     1081 gcccggtacc tcgcgaaggc cttgcaggcc aaccagataa gtgaaatcta gttccaaact
     1141 attttgtcat ttttaatttt cgtattagct tacgacgcta cacccagttc ccatctattt
     1201 tgtcactctt ccctaaataa tccttaaaaa ctccatttcc acccctccca gttcccaact
     1261 attttgtccg cccacagcgg ggcatttttc ttcctgttat gtttgggcgc tgcattaatg
     1321 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     1381 cactgacccg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagagc ttatcggcca
     1441 gcctcgcaga gcaggattcc cgttgagcac cgccaggtgc gaataaggga cagtgaagaa
     1501 ggaacacccg ctcgcgggtg ggcctacttc acctatcctg cccggctgac gccgttggat
     1561 acaccaagga aagtctacac gaaccctttg gcaaaatcct gtatatcgtg cgaaaaagga
     1621 tggatatacc gaaaaaatcg ctataatgac cccgaagcag ggttatgcag cggaaagtat
     1681 accttaagga atcccctgat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     1741 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     1801 agcatcacaa aaatcgacgc tcgagtcaga ggtggcgaaa cccgacagga ctataaagat
     1861 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     1921 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     1981 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     2041 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     2101 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     2161 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     2221 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     2281 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     2341 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     2401 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     2461 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     2521 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     2581 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     2641 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     2701 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     2761 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     2821 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     2881 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     2941 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     3001 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     3061 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     3121 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     3181 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     3241 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     3301 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     3361 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     3421 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     3481 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     3541 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc
     3601 gtttcggtga cgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
     3661 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     3721 ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
     3781 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc
     3841 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
     3901 agctagagga ccagccgcgt aacctggcaa aatcggttac ggttgagtaa taaatggatg
     3961 ccctgcgtaa gcaggtgtgg gcggacaata aagtcttaaa ctgaacaaaa tagatctaaa
     4021 ctatgacaat aaagtcttaa actagacaga atagttgtaa actgaaatca gtccagttat
     4081 gctgtgaaaa agcatactgg acttttgtta tggctaaagc aaactcttca ttttctgaag
     4141 tgcaaattgc ccgtcgtatt aaagaggggc gtggggttcg aggtcgacgg tatcgataag
     4201 ctagcttaat tagctgagct tggactcctg ttgatagatc cagtaatgac ctcagaactc
     4261 catctggatt tgttcagaac gctcggttgc cgccgggcgt tttttattgg tgagaatcca
     4321 agctagactg cgatgagtgg cagggcgggg cgtaattttt ttaaggcagt tattggtgcc
     4381 cttaaacgcc tggggtaatg actctctagc ttgaggcatc aaataaaacg aaaggctcag
     4441 tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg
     4501 acaaatccgc cgctaggagc ttgcggcccg gacgatcgaa ttggggatct tgaagtacct
     4561 attccgaagt tcctattctc tagaaagtat aggaacttca gctagagggg gcgtcaggcg
     4621 ccgggggcgg tgtccggcgg cccccagagg aactgcgcca gttcctccgg atcggtgaag
     4681 ccggagagat ccagcggggt ctcctcgaac acctcgaagt cgtgcaggaa ggtgaaggcg
     4741 agcagttcgc gggcgaagtc ctcggtccgc ttccactgcg ccccgtcgag cagcgcggcc
     4801 aggatctcgc ggtcgccccg gaaggcgttg agatgcagtt gcaccaggct gtagcgggag
     4861 tctcccgcat agacgtcggt gaagtcgacg atcccggtga cctcggtcgc ggccaggtcc
     4921 acgaagatgt tggtcccgtg caggtcgccg tggacgaacc ggggttcgcg gccggccagc
     4981 agcgtgtcca cgtccggcag ccagtcctcc aggcggtcca gcagccgggg cgagaggtag
     5041 ccccacccgc ggtggtcctc gacggtcgcc gcgcggcgtt cccgcagcag ttccgggaag
     5101 acctcggaat ggggggtgag cacggtgttc ccggtcagcg gcaccctgtg cagccggccg
     5161 agcacccggc cgagttcgcg ggccagggcg agcagcgcgt tccggtcggt cgtgccgtcc
     5221 atcgcggacc gccaggtggt gccggtcatc cggctcatca ccaggtaggg ccacggccag
     5281 gctccggtgc cgggccgcag ctcgccgcgg ccgaggaggc ggggcaccgg caccggggcg
     5341 tccgccagga ccgcgtacgc ctccgactcc gacgcgaggc tctccggacc gcaccagtgc
     5401 tcgccgaaca gcttgatcac cgggccgggc tcgccgacca gtacggggtt ggtgctctcg
     5461 ccgggcaccc gcagcaccgg cggcaccggc agcccgagct cctccagggc tcggcgggcc
     5521 agcggctccc agaattcctg gtcgttccgc aggctcgcgt aggaatcatc cgaatcaata
     5581 cggtcgagaa gtaacaggga ttcttgtgtc acagcggacc tctattcaca gggtacggg
//
