LOCUS       sequence-220092-        7336 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cloning vector for bacterial expression with Pdhfr promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7336)
  AUTHORS   Aubert DF, Flannagan RS, Valvano MA
  TITLE     A novel sensor kinase-response regulator hybrid controls biofilm 
            formation and type VI secretion system activity in Burkholderia 
            cenocepacia.
  JOURNAL   Infect Immun. 2008 May;76(5):1979-91. doi: 10.1128/IAI.01338-07. 
            Epub 2008 Mar 3.
  PUBMED    18316384
REFERENCE   2  (bases 1 to 7336)
  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"; doi: 
            "10.1128/IAI.01338-07"; journalName: "Infect Immun"; date: 
            "2008-05"; volume: "76"; issue: "5"; pages: "1979-91"
COMMENT     Sequence Label: pDA12
FEATURES             Location/Qualifiers
     source          1..7336
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(31..36,37..53,54..59)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     primer_bind     complement(487..504)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      657..743
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      835..862
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     1630..1649
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1716..1818)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(1935..1953)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     rep_origin      3211..3982
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             3983..4645
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQRRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     CDS             complement(5173..6372)
                     /codon_start=1
                     /gene="tetA"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKPNIPLIVILSTVALDAVGIGLIMPVLPGLLRDLVHSNDVTAHY
                     GILLALYALVQFACAPVLGALSDRFGRRPILLVSLAGATVDYAIMATAPFLWVLYIGRI
                     VAGITGATGAVAGAYIADITDGDERARHFGFMSACFGFGMVAGPVLGGLMGGFSPHAPF
                     FAAAALNGLNFLTGCFLLPESHKGERRPLRREALNPLASFRWARGMTVVAALMAVFFIM
                     QLVGQVPAALWVIFGEDRFHWDATTIGISLAAFGILHSLAQAMITGPVAARLGERRALM
                     LGMIADGTGYILLAFATRGWMAFPIMVLLASGGIGMPALQAMLSRQVDEERQGQLQGSL
                     AALTSLTSIVGPLLFTAIYAASITTWNGWAWIAGAALYLLCLPALRRGLWSGAGQRADR
                     "
     CDS             6478..7128
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline resistance regulatory protein"
                     /label=TetR
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKGDD"
     primer_bind     7229..7248
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 tatgctagcg atgagctcgc acgaacccag ttgacataag cctgttcggt tcgtaaactg
       61 taatgcaagt agcgtatgcg ctcacgcaac tggtccagaa ccttgaccga acgcagcggt
      121 ggtaacggcg cagtggcggt tttcatggct tgttatgact gtttttttgt acagtctatg
      181 cctcgggcat ccaagcagca agcgcgttac gccgtgggtc gatgtttgat gttatggagc
      241 agcaacgatg ttacgcagca gggcagtcgc cctaaaacaa agttaggcag ccgttgtgct
      301 ggtgctttct gatagttgtt gtggggtagg cagtcagagc tcgatttgct tgtcgccata
      361 atagattcac aagaaggatt cggaattcta catatgtcat cgacaccatg gtaccccggg
      421 atcctctaga gtcgacctgc aggcatgcaa gcttggctgt tttggcggat gagagaagat
      481 tttcagcctg atacagatta aatcagaacg cagaagcggt ctgataaaac agaatttgcc
      541 tggcggcagt agcgcggtgg tcccacctga ccccatgccg aactcagaag tgaaacgccg
      601 tagcgccgat ggtagtgtgg ggtctcccca tgcgagagta gggaactgcc aggcatcaaa
      661 taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga
      721 acgctctcct gagtaggaca aatccgccgg gagcggattt gaacgctgcg aagcaacggc
      781 ccggagggtg gcgggcagga cgcccgccat aaactgccag gcatcaaatt aagcagaagg
      841 ccatcctgac ggatggcctt tttgcgtttc tacaaactct tccactcact acacagagcc
      901 atttaaacaa catcccctcc ccctttccac cgcgtcagac gcccgtagca gcccgctacg
      961 ggctttttca tgccctgccc tagcgtccaa gcctcacggc cgcgctcggc ctctctggcg
     1021 gccttctggc gctgaggtct gcctcgtgaa gaaggtgttg ctgactcata ccaggcctga
     1081 atcgccccat catccagcca gaaagtgagg gagccacggt tgatgagagc tttgttgtag
     1141 gtggaccagt tggtgatttt gaacttttgc tttgccacgg aacggtctgc gttgtcggga
     1201 agatgcgtga tctgatcctt caactcagca aaagttcgat ttattcaaca aagccgccgt
     1261 cccgtcaagt cagcgtaatg ctctgccagt gttacaacca attaaccaat tctgattaga
     1321 aaaactcatc gagcatcaaa tgaaactgca atttattcat atcaggatta tcaataccgt
     1381 atttttgaaa aagccgtttc tgtaatgaag gagaaaactc accgaggcag ttccatagga
     1441 tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc aacatcaata caacctatta
     1501 atttcccctc gtcaaaaata aggttatcaa gtgagaaatc accatgagtg acgactgaat
     1561 ccggtgagaa tggcaaaagc taaaaaggcc gtaatatcca gctgaacggt ctggttatag
     1621 gtacattgag caactgactg aaatgcctca aaatgttctt tacgatgcca ttgggatata
     1681 tcaacggtgg tatatccagt gatttttttc tccattttag cttccttagc tcctgaaaat
     1741 ctcgataact caaaaaatac gcccggtagt gatcttattt cattatggtg aaagttggaa
     1801 cctcttacgt gccgatcaac gtctcatttt cgccaaaagt tggcccaggg cttcccggta
     1861 tcaacaggga caccaggatt tatttattct gcgaagtgat cttccgtcac aggtatttat
     1921 tcgaagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     1981 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcgcccgcg ttcctgctgg
     2041 cgctgggcct gtttctggcg ctggacttcc cgctgttccg tcagcagctt ttcgcccacg
     2101 gccttgatga tcgcggcggc cttggcctgc atatcccgat tcaacggccc cagggcgtcc
     2161 agaacgggct tcaggcgctc ccgaaggtct cgggccgtct cttgggcttg atcggccttc
     2221 ttgcgcatct cacgcgctcc tgcggcggcc tgtagggcag gctcataccc ctgccgaacc
     2281 gcttttgtca gccggtcggc cacggcttcc ggcgtctcaa cgcgctttga gattcccagc
     2341 ttttcggcca atccctgcgg tgcataggcg cgtggctcga ccgcttgcgg gctgatggtg
     2401 acgtggccca ctggtggccg ctccagggcc tcgtagaacg cctgaatgcg cgtgtgacgt
     2461 gccttgctgc cctcgatgcc ccgttgcagc cctagatcgg ccacagcggc cgcaaacgtg
     2521 gtctggtcgc gggtcatctg cgctttgttg ccgatgaact ccttggccga cagcctgccg
     2581 tcctgcgtca gcggcaccac gaacgcggtc atgtgcgggc tggtttcgtc acggtggatg
     2641 ctggccgtca cgatgcgatc cgccccgtac ttgtccgcca gccacttgtg cgccttctcg
     2701 aagaacgccg cctgctgttc ttggctggcc gacttccacc attccgggct ggccgtcatg
     2761 acgtactcga ccgccaacac agcgtccttg cgccgcttct ctggcagcaa ctcgcgcagt
     2821 cggcccatcg cttcatcggt gctgctggcc gcccagtgct cgttctctgg cgtcctgctg
     2881 gcgtcagcgt tgggcgtctc gcgctcgcgg taggcgtgct tgagactggc cgccacgttg
     2941 cccattttcg ccagcttctt gcatcgcatg atcgcgtatg ccgccatgcc tgcccctccc
     3001 ttttggtgtc caaccggctc gacgggggca gcgcaaggcg gtgcctccgg cgggccactc
     3061 aatgcttgag tatactcact agactttgct tcgcaaagtc gtgaccgcct acggcggctg
     3121 cggcgcccta cgggcttgct ctccgggctt cgccctgcgc ggtcgctgcg ctcccttgcc
     3181 agcccgtgga tatgtggacg atggccgcga gcggccaccg gctggctcgc ttcgctcggc
     3241 ccgtggacaa ccctgctgga caagctgatg gacaggctgc gcctgcccac gagcttgacc
     3301 acagggattg cccaccggct acccagcctt cgaccacata cccaccggct ccaactgcgc
     3361 ggcctgcggc cttgccccat caattttttt aattttctct ggggaaaagc ctccggcctg
     3421 cggcctgcgc gcttcgcttg ccggttggac accaagtgga aggcgggtca aggctcgcgc
     3481 agcgaccgcg cagcggcttg gccttgacgc gcctggaacg acccaagcct atgcgagtgg
     3541 gggcagtcga agggcgaagc ccgcccgcct gccccccgag cctcacggcg gcgagtgcgg
     3601 gggttccaag ggggcagcgc caccttgggc aaggccgaag gccgcgcagt cgatcaacaa
     3661 gccccggagg ggccactttt tgccggaggg ggagccgcgc cgaaggcgtg ggggaacccc
     3721 gcaggggtgc ccttctttgg gcaccaaaga actagatata gggcgaaatg cgaaagactt
     3781 aaaaatcaac aacttaaaaa aggggggtac gcaacagctc attgcggcac cccccgcaat
     3841 agctcattgc gtaggttaaa gaaaatctgt aattgactgc cacttttacg caacgcataa
     3901 ttgttgtcgc gctgccgaaa agttgcagct gattgcgcat ggtgccgcaa ccgtgcggca
     3961 cccctaccgc atggagataa gcatggccac gcagtccaga gaaatcggca ttcaagccaa
     4021 gaacaagccc ggtcactggg tgcaaacgga acgcaaagcg catgaggcgt gggccgggct
     4081 tattgcgagg aaacccacgg cggcaatgct gctgcatcac ctcgtggcgc agatgggcca
     4141 ccagaacgcc gtggtggtca gccagaagac actttccaag ctcatcggac gttctttgcg
     4201 gacggtccaa tacgcagtca aggacttggt ggccgagcgc tggatctccg tcgtgaagct
     4261 caacggcccc ggcaccgtgt cggcctacgt ggtcaatgac cgcgtggcgt ggggccagcc
     4321 ccgcgaccag ttgcgcctgt cggtgttcag tgccgccgtg gtggttgatc acgacgacca
     4381 ggacgaatcg ctgttggggc atggcgacct gcgccgcatc ccgaccctgt atccgggcga
     4441 gcagcaacta ccgaccggcc ccggcgagga gccgcccagc cagcccggca ttccgggcat
     4501 ggaaccagac ctgccagcct tgaccgaaac ggaggaatgg gaacggcgcg ggcagcggcg
     4561 cctgccgatg cccgatgagc cgtgttttct ggacgatggc gagccgttgg agccgccgac
     4621 acgggtcacg ctgccgcgcc ggtagcactt gggttgcgca gcaacccgta agtgcgctgt
     4681 tccagactat cggctgtagc cgcctcgccg ccctatacct tgtctgcctc cccgcgttgc
     4741 gtcgcggtgc atggagccgg gccacctcga cctgaatgga agccggcggc acctcgctaa
     4801 cggattcacc gtttttatca ggctctggga ggcagaataa atgatcatat cgtcaattat
     4861 tacctccacg gggagagcct gagcaaactg gcctcaggca tttgagaagc acacggtcac
     4921 actgcttccg gtagtcaata aaccggtaaa ccagcaatag acataagcgg ctatttaacg
     4981 accctgccct gaaccgacga ccgggtcgaa tttgctttcg aatttctgcc attcatccgc
     5041 ttattatcac ttattcaggc gtagcaccag gcgtttaagt cgaccaataa cctacggggt
     5101 ctgacgctca gtggaacgaa atcgatgcga agaagttgtc catattggcc acgtttccta
     5161 tcgtttccac gatcagcgat cggctcgttg ccctgcgccg ctccaaagcc cgcgacgcag
     5221 cgccggcagg cagagcaagt agagggcagc gcctgcaatc catgcccacc cgttccacgt
     5281 tgttatagaa gccgcataga tcgccgtgaa gaggaggggt ccgacgatcg aggtcaggct
     5341 ggtgagcgcc gccagtgagc cttgcagctg cccctgacgt tcctcatcca cctgcctgga
     5401 caacattgct tgcagcgccg gcattccgat gccacccgaa gcaagcagga ccatgatcgg
     5461 gaacgccatc catccccgtg tcgcgaaggc aagcaggatg tagcctgtgc cgtcggcaat
     5521 cattccgagc atgagtgccc gcctttcgcc gagccgggcg gctacagggc cggtgatcat
     5581 tgcctgggcg agtgaatgca gaatgccaaa tgcggcaagc gaaatgccga tcgtggtcgc
     5641 gtcccagtga aagcgatcct cgccgaaaat gacccaaagc gcggccggca cctgtccgac
     5701 aagttgcatg atgaagaaga ccgccatcag ggcggcgacg acggtcatgc cccgggccca
     5761 ccggaacgaa gcgagcgggt tgagagcctc ccggcgtaac ggccggcgtt cgcctttgtg
     5821 cgactccggc aaaaggaaac agcccgtcag gaaattgagg ccgttcaagg ctgccgcggc
     5881 gaagaacgga gcgtgggggg agaaaccgcc catcagccca ccgagcacag gtcccgcgac
     5941 catcccgaac ccgaaacagg cgctcatgaa gccgaagtgc cgcgcgcgct catcgccatc
     6001 agtgatatcg gcaatataag cgccggctac cgccccagtc gccccggtga tgccggccac
     6061 gatccgcccg atatagagaa cccaaaggaa aggcgctgtc gccatgatgg cgtagtcgac
     6121 agtggcgccg gccagcgaga cgagcaagat tggccgccgc ccgaaacgat ccgacagcgc
     6181 gcccagcaca ggtgcgcagg caaattgcac caacgcatac agcgccagca gaatgccata
     6241 gtgggcggtg acgtcgttcg agtgaaccag atcgcgcagg aggcccggca gcaccggcat
     6301 aatcaggccg atgccgacag cgtcgagcgc gacagtgctc agaattacga tcaggggtat
     6361 gttgggtttc acgtctggcc tccggaccag cctccgctgg tccgattgaa cgcgcggatt
     6421 ctttatcact gataagttgg tggacatatt atgtttatca gtgataaagt gtcaagcatg
     6481 acaaagttgc agccgaatac agtgatccgt gccgccctgg acctgttgaa cgaggtcggc
     6541 gtagacggtc tgacgacacg caaactggcg gaacggttgg gggttcagca gccggcgctt
     6601 tactggcact tcaggaacaa gcgggcgctg ctcgacgcac tggccgaagc catgctggcg
     6661 gagaatcata cgcattcggt gccgagagcc gacgacgact ggcgctcatt tctgatcggg
     6721 aatgcccgca gcttcaggca ggcgctgctc gcctaccgcg atggcgcgcg catccatgcc
     6781 ggcacgcgac cgggcgcacc gcagatggaa acggccgacg cgcagcttcg cttcctctgc
     6841 gaggcgggtt tttcggccgg ggacgccgtc aatgcgctga tgacaatcag ctacttcact
     6901 gttggggccg tgcttgagga gcaggccggc gacagcgatg ccggcgagcg cggcggcacc
     6961 gttgaacagg ctccgctctc gccgctgttg cgggccgcga tagacgcctt cgacgaagcc
     7021 ggtccggacg cagcgttcga gcagggactc gcggtgattg tcgatggatt ggcgaaaagg
     7081 aggctcgttg tcaggaacgt tgaaggaccg agaaagggtg acgattgatc aggaccgctg
     7141 ccggagcgca acccactcac tacagcagag ccatcgatcg atctgcactt gaacgtgtgg
     7201 cctaagcgat atcttaggat ctcccatcgg tgatgtcggc gatataggcg ccagcaaccg
     7261 cacctgtggc gccggtgatg ccggccacga tgcgtccggc gtagaggatc tgctcatgtt
     7321 tgacagctta ctcgag
//
