LOCUS       sequence-437319-        8931 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Empty backbone 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 8931)
  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 8931)
  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: pMApra-Ptet2
FEATURES             Location/Qualifiers
     source          1..8931
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..840
                     /codon_start=1
                     /gene="repA"
                     /product="replication protein A of the broad-host-range 
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /label=RSF1010 RepA
                     /translation="MATHKPINILEAFAAAPPPLDYVLPNMVAGTVGALVSPGGAGKSM
                     LALQLAAQIAGGPDLLEVGELPTGPVIYLPAEDPPTAIHHRLHALGAHLSAEERQAVAD
                     GLLIQPLIGSLPNIMAPEWFDGLKRAAEGRRLMVLDTLRRFHIEEENASGPMAQVIGRM
                     EAIAADTGCSIVFLHHASKGAAMMGAGDQQQASRGSSVLVDNIRWQSYLSSMTSAEAEE
                     WGVDDDQRRFFVRFGVSKANYGAPFADRWFRRHDGGVLKPAVLERQRKSKGVPRGEA"
     CDS             827..1678
                     /codon_start=1
                     /gene="repC"
                     /product="replication protein C of the broad-host-range 
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /label=RSF1010 RepC
                     /translation="MVKPKNKHSLSHVRHDPAHCLAPGLFRALKRGERKRSKLDVTYDY
                     GDGKRIEFSGPEPLGADDLRILQGLVAMAGPNGLVLGPEPKTEGGRQLRLFLEPKWEAV
                     TADAMVVKGSYRALAKEIGAEVDSGGALKHIQDCIERLWKVSIIAQNGRKRQGFRLLSE
                     YASDEADGRLYVALNPLIAQAVMGGGQHVRISMDEVRALDSETARLLHQRLCGWIDPGK
                     TGKASIDTLCGYVWPSEASGSTMRKRRQRVREALPELVALGWTVTEFAAGKYDITRPKA
                     AG"
     CDS             complement(1946..2572)
                     /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     2477..2496
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     promoter        2588..2643
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    2624..2642
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      2785..2871
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2963..2990
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3009..3100
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3543..3645
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(3712..3731)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             4058..4858
                     /codon_start=1
                     /gene="aac(3)IV"
                     /product="aminoglycoside 3-N-acetyltransferase type IV     
                               "
                     /label=ApmR
                     /note="confers resistance to apramycin (Paget and Davies, 
                     1996)"
                     /translation="MSSAVECNVVQYEWRKAELIGQLLNLGVTPGGVLLVHSSFRSVRP
                     LEDGPLGLIEALRAALGPGGTLVMPSWSGLDDEPFDPATSPVTPDLGVVSDTFWRLPNV
                     KRSAHPFAFAAAGPQAEQIISDPLPLPPHSPASPVARVHELDGQVLLLGVGHDANTTLH
                     LAELMAKVPYGVPRHCTILQDGKLVRVDYLENDHCCERFALADRWLKEKSLQKEGPVGH
                     AFARLIRSRDIVATALGQLGRDPLIFLHPPEAGCEECDAARQSIG"
     rep_origin      5388..5782
                     /label=RSF1010 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     RSF1010; requires the RSF1010 RepA/B/C proteins for 
                     replication (Scholz et al., 1989)"
     oriT            6124..6211
                     /label=RSF1010 oriT
                     /note="origin of transfer of the broad-host-range plasmid 
                     RSF1010 (Scholz et al., 1989)"
     CDS             7450..8421
                     /codon_start=1
                     /gene="repB"
                     /product="replication protein B of the broad-host-range 
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /label=RSF1010 RepB
                     /translation="MKNDRTLQAIGRQLKAMGCERFDIGVRDATTGQMMNREWSAAEVL
                     QNTPWLKRMNAQGNDVYIRPAEQERHGLVLVDDLSEFDLDDMKAEGREPALVVETSPKN
                     YQAWVKVADAAGGELRGQIARTLASEYDADPASADSRHYGRLAGFTNRKDKHTTRAGYQ
                     PWVLLRESKGKTATAGPALVQQAGQQIEQAQRQQEKARRLASLELPERQLSRHRRTALD
                     EYRSEMAGLVKRFGDDLSKCDFIAAQKLASRGRSAEEIGKAMAEASPALAERKPGHEAD
                     YIERTVSKVMGLPSVQLARAELARAPAPRQRGMDRGGPDFSM"
ORIGIN
        1 atggctaccc ataagcctat caatattctg gaggcgttcg cagcagcgcc gccaccgctg
       61 gactacgttt tgcccaacat ggtggccggt acggtcgggg cgctggtgtc gcccggtggt
      121 gccggtaaat ccatgctggc cctgcaactg gccgcacaga ttgcaggcgg gccggatctg
      181 ctggaggtgg gcgaactgcc caccggcccg gtgatctacc tgcccgccga agacccgccc
      241 accgccattc atcaccgcct gcacgccctt ggggcgcacc tcagcgccga ggaacggcaa
      301 gccgtggctg acggcctgct gatccagccg ctgatcggca gcctgcccaa catcatggcc
      361 ccggagtggt tcgacggcct caagcgcgcc gccgagggcc gccgcctgat ggtgctggac
      421 acgctgcgcc ggttccacat cgaggaagaa aacgccagcg gccccatggc ccaggtcatc
      481 ggtcgcatgg aggccatcgc cgccgatacc gggtgctcta tcgtgttcct gcaccatgcc
      541 agcaagggcg cggccatgat gggcgcaggc gaccagcagc aggccagccg gggcagctcg
      601 gtactggtcg ataacatccg ctggcagtcc tacctgtcga gcatgaccag cgccgaggcc
      661 gaggaatggg gtgtggacga cgaccagcgc cggttcttcg tccgcttcgg tgtgagcaag
      721 gccaactatg gcgcaccgtt cgctgatcgg tggttcaggc ggcatgacgg cggggtgctc
      781 aagcccgccg tgctggagag gcagcgcaag agcaaggggg tgccccgtgg tgaagcctaa
      841 gaacaagcac agcctcagcc acgtccggca cgacccggcg cactgtctgg cccccggcct
      901 gttccgtgcc ctcaagcggg gcgagcgcaa gcgcagcaag ctggacgtga cgtatgacta
      961 cggcgacggc aagcggatcg agttcagcgg cccggagccg ctgggcgctg atgatctgcg
     1021 catcctgcaa gggctggtgg ccatggctgg gcctaatggc ctagtgcttg gcccggaacc
     1081 caagaccgaa ggcggacggc agctccggct gttcctggaa cccaagtggg aggccgtcac
     1141 cgctgatgcc atggtggtca aaggtagcta tcgggcgctg gcaaaggaaa tcggggcaga
     1201 ggtcgatagt ggtggggcgc tcaagcacat acaggactgc atcgagcgcc tttggaaggt
     1261 atccatcatc gcccagaatg gccgcaagcg gcaggggttt cggctgctgt cggagtacgc
     1321 cagcgacgag gcggacgggc gcctgtacgt ggccctgaac cccttgatcg cgcaggccgt
     1381 catgggtggc ggccagcatg tgcgcatcag catggacgag gtgcgggcgc tggacagcga
     1441 aaccgcccgc ctgctgcacc agcggctgtg tggctggatc gaccccggca aaaccggcaa
     1501 ggcttccata gataccttgt gcggctatgt ctggccgtca gaggccagtg gttcgaccat
     1561 gcgcaagcgc cgccagcggg tgcgcgaggc gttgccggag ctggtcgcgc tgggctggac
     1621 ggtaaccgag ttcgcggcgg gcaagtacga catcacccgg cccaaggcgg caggctgacc
     1681 ccccccactc tattgtaaac aagacatttt tatcttttat attcaatggc ttattttcct
     1741 gctaattggt aataccatga aaaataccat gctcagaaaa ggcttaacaa tattttgaaa
     1801 aattgcctac tgagcgctgc cgcacagctc cataggccgc tttcctggct ttgcttccag
     1861 atgtatgctc ttctgctccc gaacgccagc aagacgtagc ccagcgcgtc ggccagcttg
     1921 caattcgcgc taacttacat taattttaag acccactttc acatttaagt tgtttttcta
     1981 atccgcatat gatcaattca aggccgaata agaaggctgg ctctgcacct tggtgatcaa
     2041 ataattcgat agcttgtcgt aataatggcg gcatactatc agtagtaggt gtttcccttt
     2101 cttctttagc gacttgatgc tcttgatctt ccaatacgca acctaaagta aaatgcccca
     2161 cagcgctgag tgcatataat gcattctcta gtgaaaaacc ttgttggcat aaaaaggcta
     2221 attgattttc gagagtttca tactgttttt ctgtaggccg tgtacctaaa tgtacttttg
     2281 ctccatcgcg atgacttagt aaagcacatc taaaactttt agcgttatta cgtaaaaaat
     2341 cttgccagct ttccccttct aaagggcaaa agtgagtatg gtgcctatct aacatctcaa
     2401 tggctaaggc gtcgagcaaa gcccgcttat tttttacatg ccaatacaat gtaggctgct
     2461 ctacacctag cttctgggcg agtttacggg ttgttaaacc ttcgattccg acctcattaa
     2521 gcagctctaa tgcgctgtta atcactttac ttttatctaa tctagacatc attaattcct
     2581 aatttttgtt gacactctat cgttgataga gttattttac cactccctat cagtgataga
     2641 gaaaatctag agacgtcgaa ttcctgcaga agcttactag tggctgaatc ccgaaacagt
     2701 tcgcaggtaa tagttagagc ctgcataacg gtttcgggat tttttatagg tctccccatg
     2761 cgagagtagg gaactgccag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc
     2821 tttcgtttta tctgttgttt gtcggtgaac gctctcctga gtaggacaaa tccgccggga
     2881 gcggatttga acgttgcgaa gcaacggccc ggagggtggc gggcaggacg cccgccataa
     2941 actgccaggc atcaaattaa gcagaaggcc atcctgacgg atggcctttt tgcgtttcta
     3001 caaactcttt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     3061 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     3121 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     3181 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     3241 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtg atgtccggcg
     3301 gtgcttttgc cgttacgcac caccccgtca gtagctgaac aggagggaca gctgatagaa
     3361 acagaagcca ctggcctcaa aaacaccatc atacactaaa tcagtaagtt ggcagcatca
     3421 cccgacgcac tttgcgccga ataaatacct gtgacggaag atcacttcgc agaataaata
     3481 aatcctggtg tccctgttga taccgggaag ccctgggcca acttttggcg aaaatgagac
     3541 gttgatcggc acgtaagagg ttccaacttt caccataatg aaataagatc actaccgggc
     3601 gtattttttg agttatcgag attttcagga gctaaggaag ctaaaatgga gaaaaaaatc
     3661 actggatata ccaccgttga tatatcccaa tggcatcgta aagaacattt tgaggcattt
     3721 cagtcagttg ctcaatgtac ctataaccag accgttcagc tggatattac ggccttttta
     3781 tcaaggcccg atccttggag cccttgccct cccgcacgat gatcgtgccg tgatcgaaat
     3841 ccagatcctt gacccgcagt tgcaaaccct cactgatccg gctcacggta actgatgccg
     3901 tatttgcagt accagcgtac ggcccacaga atgatgtcac gctgaaaatg ccggcctttg
     3961 aatgggttca tgtgcagctc catcagcaaa aggggatgat aagtttatca ccacctacta
     4021 tttgcaacag tgccgttgat cgtgctatga tcgactgatg tcatcagcgg tggagtgcaa
     4081 tgtcgtgcaa tacgaatggc gaaaagccga gctcatcggt cagcttctca accttggggt
     4141 tacccccggc ggtgtgctgc tggtccacag ctccttccgt agcgtccggc ccctcgaaga
     4201 tgggccactt ggactgatcg aggccctgcg tgctgcgctg ggtccgggag ggacgctcgt
     4261 catgccctcg tggtcaggtc tggacgacga gccgttcgat cctgccacgt cgcccgttac
     4321 accggacctt ggagttgtct ctgacacatt ctggcgcctg ccaaatgtaa agcgcagcgc
     4381 ccatccattt gcctttgcgg cagcggggcc acaggcagag cagatcatct ctgatccatt
     4441 gcccctgcca cctcactcgc ctgcaagccc ggtcgcccgt gtccatgaac tcgatgggca
     4501 ggtacttctc ctcggcgtgg gacacgatgc caacacgacg ctgcatcttg ccgagttgat
     4561 ggcaaaggtt ccctatgggg tgccgagaca ctgcaccatt cttcaggatg gcaagttggt
     4621 acgcgtcgat tatctcgaga atgaccactg ctgtgagcgc tttgccttgg cggacaggtg
     4681 gctcaaggag aagagccttc agaaggaagg tccagtcggt catgcctttg ctcggttgat
     4741 ccgctcccgc gacattgtgg cgacagccct gggtcaactg ggccgagatc cgttgatctt
     4801 cctgcatccg ccagaggcgg gatgcgaaga atgcgatgcc gctcgccagt cgattggctg
     4861 agctcatgaa aacgtggcca atatggacaa cttcttcgcc cccgttttca ccatgggcaa
     4921 atattatacg caaggcgaca aggtgctgat gccgctggcg attcaggttc atcatgccgt
     4981 ttgtgatggc ttccatgtcg gcagaatgct taatgaatta caacagtact gcgatgagtg
     5041 gcagggcggg gcgtaatttt tttaaggcag ttattgtctt caaattcccg ttgcacatag
     5101 cccggcaatt cctttccctg ctctgccata agcgcagcga atgccgggta atactcgtca
     5161 acgatctgat agagaagggt ttgctcgggt cggtggctct ggtaacgacc agtatcccga
     5221 tcccggctgg ccgtcctggc cgccacatga ggcatgttcc gcgtccttgc aatactgtgt
     5281 ttacatacag tctatcgctt agcggaaagt tcttttaccc tcagccgaaa tgcctgccgt
     5341 tgctagacat tgccagccag tgcccgtcac tcccgtacta actgtcacga acccctgcaa
     5401 taactgtcac gcccccctgc aataactgtc acgaacccct gcaataactg tcacgccccc
     5461 aaacctgcaa acccagcagg ggcgggggct ggcggggtgt tggaaaaatc catccatgat
     5521 tatctaagaa taatccacta ggcgcggtta tcagcgccct tgtggggcgc tgctgccctt
     5581 gcccaatatg cccggccaga ggccggatag ctggtctatt cgctgcgcta ggctacacac
     5641 cgccccaccg ctgcgcggca gggggaaagg cgggcaaagc ccgctaaacc ccacaccaaa
     5701 ccccgcagaa atacgctgga gcgcttttag ccgctttagc ggcctttccc cctacccgaa
     5761 gggtggggcc gcgtgtgcag ccccgcaggg cctgtctcgg tcgatcattc agcccggctc
     5821 atccttctgg cgtggcggca gaccgaacaa ggcgcggtcg tggtcgcgtt caaggtacgc
     5881 atccattgcc gccatgagcc gatcctccgg ccactcgctg ctgttcacct tggccaaaat
     5941 catggccccc accagcacct tgcgccttgt ttcgttcttg cgctcttgct gctgttccct
     6001 tgcccgcacc cgctgaattt cggcattgat tcgcgctcgt tgttcttcga gcttggccag
     6061 ccgatccgcc gccttgttgc tccccttaac catcttgaca ccccattgtt aatgtgctgt
     6121 ctcgtaggct atcatggagg cacagcggcg gcaatcccga ccctactttg taggggaggg
     6181 cgcacttacc ggtttctctt cgagaaactg gcctaacggc cacccttcgg gcggtgcgct
     6241 ctccgagggc cattgcatgg agccgaaaag caaaagcaac agcgaggcag catggcgatt
     6301 tatcacctta cggcgaaaac cggcagcagg tcgggcggcc aatcggccag ggccaaggcc
     6361 gactacatcc agcgcgaagg caagtatgcc cgcgacatgg atgaagtctt gcacgccgaa
     6421 tccgggcaca tgccggagtt cgtcgagcgg cccgccgact actgggatgc tgccgacctg
     6481 tatgaacgcg ccaatgggcg gctgttcaag gaggtcgaat ttgccctgcc ggtcgagctg
     6541 accctcgacc agcagaaggc gctggcgtcc gagttcgccc agcacctgac cggtgccgag
     6601 cgcctgccgt atacgctggc catccatgcc ggtggcggcg agaacccgca ctgccacctg
     6661 atgatctccg agcggatcaa tgacggcatc gagcggcccg ccgctcagtg gttcaagcgg
     6721 tacaacggca agaccccgga gaagggcggg gcacagaaga ccgaagcgct caagcccaag
     6781 gcatggcttg agcagacccg cgaggcatgg gccgaccatg ccaaccgggc attagagcgg
     6841 gctggccacg acgcccgcat tgaccacaga acacttgagg cgcagggcat cgagcgcctg
     6901 cccggtgttc acctggggcc gaacgtggtg gagatggaag gccggggcat ccgcaccgac
     6961 cgggcagacg tggccctgaa catcgacacc gccaacgccc agatcatcga cttacaggaa
     7021 taccgggagg caatagacca tgaacgcaat cgacagagtg aagaaatcca gaggcatcaa
     7081 cgagttagcg gagcagatcg aaccgctggc ccagagcatg gcgacactgg ccgacgaagc
     7141 ccggcaggtc atgagccaga cccagcaggc cagcgaggcg caggcggcgg agtggctgaa
     7201 agcccagcgc cagacagggg cggcatgggt ggagctggcc aaagagttgc gggaggtagc
     7261 cgccgaggtg agcagcgccg cgcagagcgc ccggagcgcg tcgcgggggt ggcactggaa
     7321 gctatggcta accgtgatgc tggcttccat gatgcctacg gtggtgctgc tgatcgcatc
     7381 gttgctcttg ctcgacctga cgccactgac aaccgaggac ggctcgatct ggctgcgctt
     7441 ggtggcccga tgaagaacga caggactttg caggccatag gccgacagct caaggccatg
     7501 ggctgtgagc gcttcgatat cggcgtcagg gacgccacca ccggccagat gatgaaccgg
     7561 gaatggtcag ccgccgaagt gctccagaac acgccatggc tcaagcggat gaatgcccag
     7621 ggcaatgacg tgtatatcag gcccgccgag caggagcggc atggtctggt gctggtggac
     7681 gacctcagcg agtttgacct ggatgacatg aaagccgagg gccgggagcc tgccctggta
     7741 gtggaaacca gcccgaagaa ctatcaggca tgggtcaagg tggccgacgc cgcaggcggt
     7801 gaacttcggg ggcagattgc ccggacgctg gccagcgagt acgacgccga cccggccagc
     7861 gccgacagcc gccactatgg ccgcttggcg ggcttcacca accgcaagga caagcacacc
     7921 acccgcgccg gttatcagcc gtgggtgctg ctgcgtgaat ccaagggcaa gaccgccacc
     7981 gctggcccgg cgctggtgca gcaggctggc cagcagatcg agcaggccca gcggcagcag
     8041 gagaaggccc gcaggctggc cagcctcgaa ctgcccgagc ggcagcttag ccgccaccgg
     8101 cgcacggcgc tggacgagta ccgcagcgag atggccgggc tggtcaagcg cttcggtgat
     8161 gacctcagca agtgcgactt tatcgccgcg cagaagctgg ccagccgggg ccgcagtgcc
     8221 gaggaaatcg gcaaggccat ggccgaggcc agcccagcgc tggcagagcg caagcccggc
     8281 cacgaagcgg attacatcga gcgcaccgtc agcaaggtca tgggtctgcc cagcgtccag
     8341 cttgcgcggg ccgagctggc acgggcaccg gcaccccgcc agcgaggcat ggacaggggc
     8401 gggccagatt tcagcatgta gtgcttgcgt tggtactcac gcctgttata ctatgagtac
     8461 tcacgcacag aagggggttt tatggaatac gaaaaaagcg cttcagggtc ggtctacctg
     8521 atcaaaagtg acaagggcta ttggttgccc ggtggctttg gttatacgtc aaacaaggcc
     8581 gaggctggcc gcttttcagt cgctgatatg gccagcctta accttgacgg ctgcaccttg
     8641 tccttgttcc gcgaagacaa gcctttcggc cccggcaagt ttctcggtga ctgatatgaa
     8701 agaccaaaag gacaagcaga ccggcgacct gctggccagc cctgacgctg tacgccaagc
     8761 gcgatatgcc gagcgcatga aggccaaagg gatgcgtcag cgcaagttct ggctgaccga
     8821 cgacgaatac gaggcgctgc gcgagtgcct ggaagaactc agagcggcgc agggcggggg
     8881 tagtgacccc gccagcgcct aaccaccaac tgcctgcaaa ggaggcaatc a
//
