LOCUS       sequence-434355-        8907 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Empty backbone 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 8907)
  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 8907)
  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-Ptet1
FEATURES             Location/Qualifiers
     source          1..8907
                     /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"
     terminator      2761..2847
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2939..2966
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2985..3076
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3519..3621
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(3688..3707)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             4034..4834
                     /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      5364..5758
                     /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            6100..6187
                     /label=RSF1010 oriT
                     /note="origin of transfer of the broad-host-range plasmid 
                     RSF1010 (Scholz et al., 1989)"
     CDS             7426..8397
                     /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 ctttaaagac gtcgaattcc
     2641 tgcagaagct tactagtggc tgaatcccga aacagttcgc aggtaatagt tagagcctgc
     2701 ataacggttt cgggattttt tataggtctc cccatgcgag agtagggaac tgccaggcat
     2761 caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg
     2821 gtgaacgctc tcctgagtag gacaaatccg ccgggagcgg atttgaacgt tgcgaagcaa
     2881 cggcccggag ggtggcgggc aggacgcccg ccataaactg ccaggcatca aattaagcag
     2941 aaggccatcc tgacggatgg cctttttgcg tttctacaaa ctcttttgtt tatttttcta
     3001 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     3061 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     3121 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     3181 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     3241 tgagagtttt cgccccgaag aacgtgatgt ccggcggtgc ttttgccgtt acgcaccacc
     3301 ccgtcagtag ctgaacagga gggacagctg atagaaacag aagccactgg cctcaaaaac
     3361 accatcatac actaaatcag taagttggca gcatcacccg acgcactttg cgccgaataa
     3421 atacctgtga cggaagatca cttcgcagaa taaataaatc ctggtgtccc tgttgatacc
     3481 gggaagccct gggccaactt ttggcgaaaa tgagacgttg atcggcacgt aagaggttcc
     3541 aactttcacc ataatgaaat aagatcacta ccgggcgtat tttttgagtt atcgagattt
     3601 tcaggagcta aggaagctaa aatggagaaa aaaatcactg gatataccac cgttgatata
     3661 tcccaatggc atcgtaaaga acattttgag gcatttcagt cagttgctca atgtacctat
     3721 aaccagaccg ttcagctgga tattacggcc tttttatcaa ggcccgatcc ttggagccct
     3781 tgccctcccg cacgatgatc gtgccgtgat cgaaatccag atccttgacc cgcagttgca
     3841 aaccctcact gatccggctc acggtaactg atgccgtatt tgcagtacca gcgtacggcc
     3901 cacagaatga tgtcacgctg aaaatgccgg cctttgaatg ggttcatgtg cagctccatc
     3961 agcaaaaggg gatgataagt ttatcaccac ctactatttg caacagtgcc gttgatcgtg
     4021 ctatgatcga ctgatgtcat cagcggtgga gtgcaatgtc gtgcaatacg aatggcgaaa
     4081 agccgagctc atcggtcagc ttctcaacct tggggttacc cccggcggtg tgctgctggt
     4141 ccacagctcc ttccgtagcg tccggcccct cgaagatggg ccacttggac tgatcgaggc
     4201 cctgcgtgct gcgctgggtc cgggagggac gctcgtcatg ccctcgtggt caggtctgga
     4261 cgacgagccg ttcgatcctg ccacgtcgcc cgttacaccg gaccttggag ttgtctctga
     4321 cacattctgg cgcctgccaa atgtaaagcg cagcgcccat ccatttgcct ttgcggcagc
     4381 ggggccacag gcagagcaga tcatctctga tccattgccc ctgccacctc actcgcctgc
     4441 aagcccggtc gcccgtgtcc atgaactcga tgggcaggta cttctcctcg gcgtgggaca
     4501 cgatgccaac acgacgctgc atcttgccga gttgatggca aaggttccct atggggtgcc
     4561 gagacactgc accattcttc aggatggcaa gttggtacgc gtcgattatc tcgagaatga
     4621 ccactgctgt gagcgctttg ccttggcgga caggtggctc aaggagaaga gccttcagaa
     4681 ggaaggtcca gtcggtcatg cctttgctcg gttgatccgc tcccgcgaca ttgtggcgac
     4741 agccctgggt caactgggcc gagatccgtt gatcttcctg catccgccag aggcgggatg
     4801 cgaagaatgc gatgccgctc gccagtcgat tggctgagct catgaaaacg tggccaatat
     4861 ggacaacttc ttcgcccccg ttttcaccat gggcaaatat tatacgcaag gcgacaaggt
     4921 gctgatgccg ctggcgattc aggttcatca tgccgtttgt gatggcttcc atgtcggcag
     4981 aatgcttaat gaattacaac agtactgcga tgagtggcag ggcggggcgt aattttttta
     5041 aggcagttat tgtcttcaaa ttcccgttgc acatagcccg gcaattcctt tccctgctct
     5101 gccataagcg cagcgaatgc cgggtaatac tcgtcaacga tctgatagag aagggtttgc
     5161 tcgggtcggt ggctctggta acgaccagta tcccgatccc ggctggccgt cctggccgcc
     5221 acatgaggca tgttccgcgt ccttgcaata ctgtgtttac atacagtcta tcgcttagcg
     5281 gaaagttctt ttaccctcag ccgaaatgcc tgccgttgct agacattgcc agccagtgcc
     5341 cgtcactccc gtactaactg tcacgaaccc ctgcaataac tgtcacgccc ccctgcaata
     5401 actgtcacga acccctgcaa taactgtcac gcccccaaac ctgcaaaccc agcaggggcg
     5461 ggggctggcg gggtgttgga aaaatccatc catgattatc taagaataat ccactaggcg
     5521 cggttatcag cgcccttgtg gggcgctgct gcccttgccc aatatgcccg gccagaggcc
     5581 ggatagctgg tctattcgct gcgctaggct acacaccgcc ccaccgctgc gcggcagggg
     5641 gaaaggcggg caaagcccgc taaaccccac accaaacccc gcagaaatac gctggagcgc
     5701 ttttagccgc tttagcggcc tttcccccta cccgaagggt ggggccgcgt gtgcagcccc
     5761 gcagggcctg tctcggtcga tcattcagcc cggctcatcc ttctggcgtg gcggcagacc
     5821 gaacaaggcg cggtcgtggt cgcgttcaag gtacgcatcc attgccgcca tgagccgatc
     5881 ctccggccac tcgctgctgt tcaccttggc caaaatcatg gcccccacca gcaccttgcg
     5941 ccttgtttcg ttcttgcgct cttgctgctg ttcccttgcc cgcacccgct gaatttcggc
     6001 attgattcgc gctcgttgtt cttcgagctt ggccagccga tccgccgcct tgttgctccc
     6061 cttaaccatc ttgacacccc attgttaatg tgctgtctcg taggctatca tggaggcaca
     6121 gcggcggcaa tcccgaccct actttgtagg ggagggcgca cttaccggtt tctcttcgag
     6181 aaactggcct aacggccacc cttcgggcgg tgcgctctcc gagggccatt gcatggagcc
     6241 gaaaagcaaa agcaacagcg aggcagcatg gcgatttatc accttacggc gaaaaccggc
     6301 agcaggtcgg gcggccaatc ggccagggcc aaggccgact acatccagcg cgaaggcaag
     6361 tatgcccgcg acatggatga agtcttgcac gccgaatccg ggcacatgcc ggagttcgtc
     6421 gagcggcccg ccgactactg ggatgctgcc gacctgtatg aacgcgccaa tgggcggctg
     6481 ttcaaggagg tcgaatttgc cctgccggtc gagctgaccc tcgaccagca gaaggcgctg
     6541 gcgtccgagt tcgcccagca cctgaccggt gccgagcgcc tgccgtatac gctggccatc
     6601 catgccggtg gcggcgagaa cccgcactgc cacctgatga tctccgagcg gatcaatgac
     6661 ggcatcgagc ggcccgccgc tcagtggttc aagcggtaca acggcaagac cccggagaag
     6721 ggcggggcac agaagaccga agcgctcaag cccaaggcat ggcttgagca gacccgcgag
     6781 gcatgggccg accatgccaa ccgggcatta gagcgggctg gccacgacgc ccgcattgac
     6841 cacagaacac ttgaggcgca gggcatcgag cgcctgcccg gtgttcacct ggggccgaac
     6901 gtggtggaga tggaaggccg gggcatccgc accgaccggg cagacgtggc cctgaacatc
     6961 gacaccgcca acgcccagat catcgactta caggaatacc gggaggcaat agaccatgaa
     7021 cgcaatcgac agagtgaaga aatccagagg catcaacgag ttagcggagc agatcgaacc
     7081 gctggcccag agcatggcga cactggccga cgaagcccgg caggtcatga gccagaccca
     7141 gcaggccagc gaggcgcagg cggcggagtg gctgaaagcc cagcgccaga caggggcggc
     7201 atgggtggag ctggccaaag agttgcggga ggtagccgcc gaggtgagca gcgccgcgca
     7261 gagcgcccgg agcgcgtcgc gggggtggca ctggaagcta tggctaaccg tgatgctggc
     7321 ttccatgatg cctacggtgg tgctgctgat cgcatcgttg ctcttgctcg acctgacgcc
     7381 actgacaacc gaggacggct cgatctggct gcgcttggtg gcccgatgaa gaacgacagg
     7441 actttgcagg ccataggccg acagctcaag gccatgggct gtgagcgctt cgatatcggc
     7501 gtcagggacg ccaccaccgg ccagatgatg aaccgggaat ggtcagccgc cgaagtgctc
     7561 cagaacacgc catggctcaa gcggatgaat gcccagggca atgacgtgta tatcaggccc
     7621 gccgagcagg agcggcatgg tctggtgctg gtggacgacc tcagcgagtt tgacctggat
     7681 gacatgaaag ccgagggccg ggagcctgcc ctggtagtgg aaaccagccc gaagaactat
     7741 caggcatggg tcaaggtggc cgacgccgca ggcggtgaac ttcgggggca gattgcccgg
     7801 acgctggcca gcgagtacga cgccgacccg gccagcgccg acagccgcca ctatggccgc
     7861 ttggcgggct tcaccaaccg caaggacaag cacaccaccc gcgccggtta tcagccgtgg
     7921 gtgctgctgc gtgaatccaa gggcaagacc gccaccgctg gcccggcgct ggtgcagcag
     7981 gctggccagc agatcgagca ggcccagcgg cagcaggaga aggcccgcag gctggccagc
     8041 ctcgaactgc ccgagcggca gcttagccgc caccggcgca cggcgctgga cgagtaccgc
     8101 agcgagatgg ccgggctggt caagcgcttc ggtgatgacc tcagcaagtg cgactttatc
     8161 gccgcgcaga agctggccag ccggggccgc agtgccgagg aaatcggcaa ggccatggcc
     8221 gaggccagcc cagcgctggc agagcgcaag cccggccacg aagcggatta catcgagcgc
     8281 accgtcagca aggtcatggg tctgcccagc gtccagcttg cgcgggccga gctggcacgg
     8341 gcaccggcac cccgccagcg aggcatggac aggggcgggc cagatttcag catgtagtgc
     8401 ttgcgttggt actcacgcct gttatactat gagtactcac gcacagaagg gggttttatg
     8461 gaatacgaaa aaagcgcttc agggtcggtc tacctgatca aaagtgacaa gggctattgg
     8521 ttgcccggtg gctttggtta tacgtcaaac aaggccgagg ctggccgctt ttcagtcgct
     8581 gatatggcca gccttaacct tgacggctgc accttgtcct tgttccgcga agacaagcct
     8641 ttcggccccg gcaagtttct cggtgactga tatgaaagac caaaaggaca agcagaccgg
     8701 cgacctgctg gccagccctg acgctgtacg ccaagcgcga tatgccgagc gcatgaaggc
     8761 caaagggatg cgtcagcgca agttctggct gaccgacgac gaatacgagg cgctgcgcga
     8821 gtgcctggaa gaactcagag cggcgcaggg cgggggtagt gaccccgcca gcgcctaacc
     8881 accaactgcc tgcaaaggag gcaatca
//
