LOCUS       sequence-432233-        5607 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Empty backbone for Tn7-site insertion with tetR-PtetA expression 
            system with single tetR operator site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5607)
  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 5607)
  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-Ptet1
FEATURES             Location/Qualifiers
     source          1..5607
                     /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        join(984..989,990..1007,1008..1012,1013..1018)
                     /label=tetA promoter
                     /note="contains a tet operator sequence that enables 
                     repression by TetR"
     mobile_element  1079..1244
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     oriT            1403..1511
                     /note="incP origin of transfer"
     rep_origin      complement(1743..2331)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1823..1842)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2502..3293,3294..3362))
                     /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     3125..3144
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3363..3467)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3535..3553
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3591..3613)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3713..3732
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4146..4162
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     4147..4163
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      4191..4285
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      4388..4474
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     protein_bind    4520..4567
                     /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 ccctgcagga attcctcgag aagcttgggc ccggtacctc gcgaaggcct tgcaggccaa
     1081 ccagataagt gaaatctagt tccaaactat tttgtcattt ttaattttcg tattagctta
     1141 cgacgctaca cccagttccc atctattttg tcactcttcc ctaaataatc cttaaaaact
     1201 ccatttccac ccctcccagt tcccaactat tttgtccgcc cacagcgggg catttttctt
     1261 cctgttatgt ttgggcgctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     1321 gtattgggcg ctcttccgct tcctcgctca ctgacccgct gcgctcggtc gttcggctgc
     1381 ggcgagcggt atcagagctt atcggccagc ctcgcagagc aggattcccg ttgagcaccg
     1441 ccaggtgcga ataagggaca gtgaagaagg aacacccgct cgcgggtggg cctacttcac
     1501 ctatcctgcc cggctgacgc cgttggatac accaaggaaa gtctacacga accctttggc
     1561 aaaatcctgt atatcgtgcg aaaaaggatg gatataccga aaaaatcgct ataatgaccc
     1621 cgaagcaggg ttatgcagcg gaaagtatac cttaaggaat cccctgataa cgcaggaaag
     1681 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     1741 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc gagtcagagg
     1801 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     1861 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     1921 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     1981 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     2041 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     2101 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     2161 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     2221 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     2281 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     2341 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     2401 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     2461 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     2521 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     2581 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     2641 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     2701 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     2761 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     2821 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     2881 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     2941 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     3001 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     3061 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     3121 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     3181 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     3241 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     3301 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     3361 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     3421 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     3481 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     3541 cgtatcacga ggccctttcg tctcgcgcgt ttcggtgacg acggtgaaaa cctctgacac
     3601 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
     3661 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca
     3721 gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg
     3781 agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga
     3841 tcggtgcggg cctcttcgct attacgccag ctagaggacc agccgcgtaa cctggcaaaa
     3901 tcggttacgg ttgagtaata aatggatgcc ctgcgtaagc aggtgtgggc ggacaataaa
     3961 gtcttaaact gaacaaaata gatctaaact atgacaataa agtcttaaac tagacagaat
     4021 agttgtaaac tgaaatcagt ccagttatgc tgtgaaaaag catactggac ttttgttatg
     4081 gctaaagcaa actcttcatt ttctgaagtg caaattgccc gtcgtattaa agaggggcgt
     4141 ggggttcgag gtcgacggta tcgataagct agcttaatta gctgagcttg gactcctgtt
     4201 gatagatcca gtaatgacct cagaactcca tctggatttg ttcagaacgc tcggttgccg
     4261 ccgggcgttt tttattggtg agaatccaag ctagactgcg atgagtggca gggcggggcg
     4321 taattttttt aaggcagtta ttggtgccct taaacgcctg gggtaatgac tctctagctt
     4381 gaggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg
     4441 tttgtcggtg aacgctctcc tgagtaggac aaatccgccg ctaggagctt gcggcccgga
     4501 cgatcgaatt ggggatcttg aagtacctat tccgaagttc ctattctcta gaaagtatag
     4561 gaacttcagc tagagggggc gtcaggcgcc gggggcggtg tccggcggcc cccagaggaa
     4621 ctgcgccagt tcctccggat cggtgaagcc ggagagatcc agcggggtct cctcgaacac
     4681 ctcgaagtcg tgcaggaagg tgaaggcgag cagttcgcgg gcgaagtcct cggtccgctt
     4741 ccactgcgcc ccgtcgagca gcgcggccag gatctcgcgg tcgccccgga aggcgttgag
     4801 atgcagttgc accaggctgt agcgggagtc tcccgcatag acgtcggtga agtcgacgat
     4861 cccggtgacc tcggtcgcgg ccaggtccac gaagatgttg gtcccgtgca ggtcgccgtg
     4921 gacgaaccgg ggttcgcggc cggccagcag cgtgtccacg tccggcagcc agtcctccag
     4981 gcggtccagc agccggggcg agaggtagcc ccacccgcgg tggtcctcga cggtcgccgc
     5041 gcggcgttcc cgcagcagtt ccgggaagac ctcggaatgg ggggtgagca cggtgttccc
     5101 ggtcagcggc accctgtgca gccggccgag cacccggccg agttcgcggg ccagggcgag
     5161 cagcgcgttc cggtcggtcg tgccgtccat cgcggaccgc caggtggtgc cggtcatccg
     5221 gctcatcacc aggtagggcc acggccaggc tccggtgccg ggccgcagct cgccgcggcc
     5281 gaggaggcgg ggcaccggca ccggggcgtc cgccaggacc gcgtacgcct ccgactccga
     5341 cgcgaggctc tccggaccgc accagtgctc gccgaacagc ttgatcaccg ggccgggctc
     5401 gccgaccagt acggggttgg tgctctcgcc gggcacccgc agcaccggcg gcaccggcag
     5461 cccgagctcc tccagggctc ggcgggccag cggctcccag aattcctggt cgttccgcag
     5521 gctcgcgtag gaatcatccg aatcaatacg gtcgagaagt aacagggatt cttgtgtcac
     5581 agcggacctc tattcacagg gtacggg
//
