LOCUS       sequence-214625-        7332 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 7332)
  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 7332)
  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..7332
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(51..701)
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline resistance regulatory protein"
                     /label=TetR
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKGDD"
     CDS             807..2006
                     /codon_start=1
                     /gene="tetA"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKPNIPLIVILSTVALDAVGIGLIMPVLPGLLRDLVHSNDVTAHY
                     GILLALYALVQFACAPVLGALSDRFGRRPILLVSLAGATVDYAIMATAPFLWVLYIGRI
                     VAGITGATGAVAGAYIADITDGDERARHFGFMSACFGFGMVAGPVLGGLMGGFSPHAPF
                     FAAAALNGLNFLTGCFLLPESHKGERRPLRREALNPLASFRWARGMTVVAALMAVFFIM
                     QLVGQVPAALWVIFGEDRFHWDATTIGISLAAFGILHSLAQAMITGPVAARLGERRALM
                     LGMIADGTGYILLAFATRGWMAFPIMVLLASGGIGMPALQAMLSRQVDEERQGQLQGSL
                     AALTSLTSIVGPLLFTAIYAASITTWNGWAWIAGAALYLLCLPALRRGLWSGAGQRADR
                     "
     primer_bind     2105..2124
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        join(2243..2248,2249..2265,2266..2271)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     primer_bind     complement(2699..2716)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2869..2955
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3047..3074
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     3843..3862
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(3929..4031)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(4148..4166)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     rep_origin      5424..6193
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             6194..6856
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
ORIGIN
        1 atcgatggct ctgctgtagt gagtgggttg cgctccggca gcggtcctga tcaatcgtca
       61 ccctttctcg gtccttcaac gttcctgaca acgagcctcc ttttcgccaa tccatcgaca
      121 atcaccgcga gtccctgctc gaacgctgcg tccggaccgg cttcgtcgaa ggcgtctatc
      181 gcggcccgca acagcggcga gagcggagcc tgttcaacgg tgccgccgcg ctcgccggca
      241 tcgctgtcgc cggcctgctc ctcaagcacg gccccaacag tgaagtagct gattgtcatc
      301 agcgcattga cggcgtcccc ggccgaaaaa cccgcctcgc agaggaagcg aagctgcgcg
      361 tcggccgttt ccatctgcgg tgcgcccggt cgcgtgccgg catggatgcg cgcgccatcg
      421 cggtaggcga gcagcgcctg cctgaagctg cgggcattcc cgatcagaaa tgagcgccag
      481 tcgtcgtcgg ctctcggcac cgaatgcgta tgattctccg ccagcatggc ttcggccagt
      541 gcgtcgagca gcgcccgctt gttcctgaag tgccagtaaa gcgccggctg ctgaaccccc
      601 aaccgttccg ccagtttgcg tgtcgtcaga ccgtctacgc cgacctcgtt caacaggtcc
      661 agggcggcac ggatcactgt attcggctgc aactttgtca tgcttgacac tttatcactg
      721 ataaacataa tatgtccacc aacttatcag tgataaagaa tccgcgcgtt caatcggacc
      781 agcggaggct ggtccggagg ccagacgtga aacccaacat acccctgatc gtaattctga
      841 gcactgtcgc gctcgacgct gtcggcatcg gcctgattat gccggtgctg ccgggcctcc
      901 tgcgcgatct ggttcactcg aacgacgtca ccgcccacta tggcattctg ctggcgctgt
      961 atgcgttggt gcaatttgcc tgcgcacctg tgctgggcgc gctgtcggat cgtttcgggc
     1021 ggcggccaat cttgctcgtc tcgctggccg gcgccactgt cgactacgcc atcatggcga
     1081 cagcgccttt cctttgggtt ctctatatcg ggcggatcgt ggccggcatc accggggcga
     1141 ctggggcggt agccggcgct tatattgccg atatcactga tggcgatgag cgcgcgcggc
     1201 acttcggctt catgagcgcc tgtttcgggt tcgggatggt cgcgggacct gtgctcggtg
     1261 ggctgatggg cggtttctcc ccccacgctc cgttcttcgc cgcggcagcc ttgaacggcc
     1321 tcaatttcct gacgggctgt ttccttttgc cggagtcgca caaaggcgaa cgccggccgt
     1381 tacgccggga ggctctcaac ccgctcgctt cgttccggtg ggcccggggc atgaccgtcg
     1441 tcgccgccct gatggcggtc ttcttcatca tgcaacttgt cggacaggtg ccggccgcgc
     1501 tttgggtcat tttcggcgag gatcgctttc actgggacgc gaccacgatc ggcatttcgc
     1561 ttgccgcatt tggcattctg cattcactcg cccaggcaat gatcaccggc cctgtagccg
     1621 cccggctcgg cgaaaggcgg gcactcatgc tcggaatgat tgccgacggc acaggctaca
     1681 tcctgcttgc cttcgcgaca cggggatgga tggcgttccc gatcatggtc ctgcttgctt
     1741 cgggtggcat cggaatgccg gcgctgcaag caatgttgtc caggcaggtg gatgaggaac
     1801 gtcaggggca gctgcaaggc tcactggcgg cgctcaccag cctgacctcg atcgtcggac
     1861 ccctcctctt cacggcgatc tatgcggctt ctataacaac gtggaacggg tgggcatgga
     1921 ttgcaggcgc tgccctctac ttgctctgcc tgccggcgct gcgtcgcggg ctttggagcg
     1981 gcgcagggca acgagccgat cgctgatcgt ggaaacgata ggaaacgtgg ccaatatgga
     2041 caacttcttc gcatcgatct gcacttgaac gtgtggccta agcgatatct taggatctcc
     2101 catcggtgat gtcggcgata taggcgccag caaccgcacc tgtggcgccg gtgatgccgg
     2161 ccacgatgcg tccggcgtag aggatctgct catgtttgac agcttactcg agtatgctag
     2221 cgatgagctc gcacgaaccc agttgacata agcctgttcg gttcgtaaac tgtaatgcaa
     2281 gtagcgtatg cgctcacgca actggtccag aaccttgacc gaacgcagcg gtggtaacgg
     2341 cgcagtggcg gttttcatgg cttgttatga ctgttttttt gtacagtcta tgcctcgggc
     2401 atccaagcag caagcgcgtt acgccgtggg tcgatgtttg atgttatgga gcagcaacga
     2461 tgttacgcag cagggcagtc gccctaaaac aaagttaggc agccgttgtg ctggtgcttt
     2521 ctgatagttg ttgtggggta ggcagtcaga gctcgatttg cttgtcgcca taatagattc
     2581 acaagaagga ttcggaattc tacatatgtc atcgacacca tggtacccgg ggatcctcta
     2641 gagtcgacct gcaggcatgc aagcttggct gttttggcgg atgagagaag attttcagcc
     2701 tgatacagat taaatcagaa cgcagaagcg gtctgataaa acagaatttg cctggcggca
     2761 gtagcgcggt ggtcccacct gaccccatgc cgaactcaga agtgaaacgc cgtagcgccg
     2821 atggtagtgt ggggtctccc catgcgagag tagggaactg ccaggcatca aataaaacga
     2881 aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc
     2941 ctgagtagga caaatccgcc gggagcggat ttgaacgctg cgaagcaacg gcccggaggg
     3001 tggcgggcag gacgcccgcc ataaactgcc aggcatcaaa ttaagcagaa ggccatcctg
     3061 acggatggcc tttttgcgtt tctacaaact cttccactca ctacagcaga gccatttaaa
     3121 caacatcccc tccccctttc caccgcgtca gacgcccgta gcagcccgct acgggctttt
     3181 tcatgccctg ccctagcgtc caagcctcac ggccgcgctc ggcctctctg gcggccttct
     3241 ggcgctgagg tctgcctcgt gaagaaggtg ttgctgactc ataccaggcc tgaatcgccc
     3301 catcatccag ccagaaagtg agggagccac ggttgatgag agctttgttg taggtggacc
     3361 agttggtgat tttgaacttt tgctttgcca cggaacggtc tgcgttgtcg ggaagatgcg
     3421 tgatctgatc cttcaactca gcaaaagttc gatttattca acaaagccgc cgtcccgtca
     3481 agtcagcgta atgctctgcc agtgttacaa ccaattaacc aattctgatt agaaaaactc
     3541 atcgagcatc aaatgaaact gcaatttatt catatcagga ttatcaatac catatttttg
     3601 aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg cagttccata ggatggcaag
     3661 atcctggtat cggtctgcga ttccgactcg tccaacatca atacaaccta ttaatttccc
     3721 ctcgtcaaaa ataaggttat caagcgagaa atcaccatga gtgacgactg aatccggtga
     3781 gaatggcaaa agctaaaaag gccgtaatat ccagctgaac ggtctggtta taggtacatt
     3841 gagcaactga ctgaaatgcc tcaaaatgtt ctttacgatg ccattgggat atatcaacgg
     3901 tggtatatcc agtgattttt ttctccattt tagcttcctt agctcctgaa aatctcgata
     3961 actcaaaaaa tacgcccggt agtgatctta tttcattatg gtgaaagttg gaacctctta
     4021 cgtgccgatc aacgtctcat tttcgccaaa agttggccca gggcttcccg gtatcaacag
     4081 ggacaccagg atttatttat tctgcgaagt gatcttccgt cacaggtatt tattcgaaga
     4141 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     4201 tagacgtcag gtggcacttt tcggggaaat gtgcgcgccc gcgttcctgc tggcgctggg
     4261 cctgtttctg gcgctggact tcccgctgtt ccgtcagcag cttttcgccc acggccttga
     4321 tgatcgcggc ggccttggcc tgcatatccc gattcaacgg ccccagggcg tccagaacgg
     4381 gcttcaggcg ctcccgaagg tctcgggccg tctcttgggc ttgatcggcc ttcttgcgca
     4441 tctcacgcgc tcctgcggcg gcctgtaggg caggctcata cccctgccga accgcttttg
     4501 tcagccggtc ggccacggct tccggcgtct caacgcgctt tgagattccc agcttttcgg
     4561 ccaatccctg cggtgcatag gcgcgtggct cgaccgcttg cgggctgatg gtgacgtggc
     4621 ccactggtgg ccgctccagg gcctcgtaga acgcctgaat gcgcgtgtga cgtgccttgc
     4681 tgccctcgat gccccgttgc agccctagat cggccacagc ggccgcaaac gtggtctggt
     4741 cgcgggtcat ctgcgctttg ttgccgatga actccttggc cgacagcctg ccgtcctgcg
     4801 tcagcggcac cacgaacgcg gtcatgtgcg ggctggtttc gtcacggtgg atgctggccg
     4861 tcacgatgcg atccgccccg tacttgtccg ccagccactt gtgcgccttc tcgaagaacg
     4921 ccgcctgctg ttcttggctg gccgacttcc accattccgg gctggccgtc atgacgtact
     4981 cgaccgccaa cacagcgtcc ttgcgccgct tctctggcag caactcgcgc agtcggccca
     5041 tcgcttcatc ggtgctgctg gccgcccagt gctcgttctc tggcgtcctg ctggcgtcag
     5101 cgttgggcgt ctcgcgctcg cggtaggcgt gcttgagact ggccgccacg ttgcccattt
     5161 tcgccagctt cttgcatcgc atgatcgcgt atgccgccat gcctgcccct cccttttggt
     5221 gtccaaccgg ctcgacgggg gcagcgcaag gcggtgcctc cggcgggcca ctcaatgctt
     5281 gagtatactc actagacttt gcttcgcaaa gtcgtgaccg cctacggcgg ctgcggcgcc
     5341 ctacgggctt gctctccggg cttcgccctg cgcggtcgct gcgctccctt gccagcccgt
     5401 ggatatgtgg acgatggccg cgagcggcca ccggctggct cgcttcgctc ggcccgtgga
     5461 caaccctgct ggacaagctg atggacaggc tgcgcctgcc cacgagcttg accacaggga
     5521 ttgcccaccg gctacccagc cttcgaccac atacccaccg gctccaactg cgcggcctgc
     5581 ggccttgccc catcaatttt tttaattttc tctggggaaa agcctccggc ctgcggcctg
     5641 cgcgcttcgc ttgccggttg gacaccaagt ggaaggcggg tcaaggctcg cgcagcgacc
     5701 gcgcagcggc ttggccttga cgcgcctgga acgacccaag cctatgcgag tgggggcagt
     5761 cgaaggcgaa gcccgcccgc ctgccccccg agcctcacgg cggcgagtgc gggggttcca
     5821 agggggcagc gccaccttgg gcaaggccga aggccgcgca gtcgatcaac aagccccgga
     5881 ggggccactt tttgccggag ggggagccgc gccgaaggcg tgggggaacc ccgcaggggt
     5941 gcccttcttt gggcaccaaa gaactagata tagggcgaaa tgcgaaagac ttaaaaatca
     6001 acaacttaaa aaaggggggt acgcaacagc tcattgcggc accccccgca atagctcatt
     6061 gcgtaggtta aagaaaatct gtaattgact gccactttta cgcaacgcat aattgttgtc
     6121 gcgctgccga aaagttgcag ctgattgcgc atggtgccgc aaccgtgcgg caccctaccg
     6181 catggagata agcatggcca cgcagtccag agaaatcggc attcaagcca agaacaagcc
     6241 cggtcactgg gtgcaaacgg aacgcaaagc gcatgaggcg tgggccgggc ttattgcgag
     6301 gaaacccacg gcggcaatgc tgctgcatca cctcgtggcg cagatgggcc accagaacgc
     6361 cgtggtggtc agccagaaga cactttccaa gctcatcgga cgttctttgc ggacggtcca
     6421 atacgcagtc aaggacttgg tggccgagcg ctggatctcc gtcgtgaagc tcaacggccc
     6481 cggcaccgtg tcggcctacg tggtcaatga ccgcgtggcg tggggccagc cccgcgacca
     6541 gttgcgcctg tcggtgttca gtgccgccgt ggtggttgat cacgacgacc aggacgaatc
     6601 gctgttgggg catggcgacc tgcgccgcat cccgaccctg tatccgggcg agcagcaact
     6661 accgaccggc cccggcgagg agccgcccag ccagcccggc attccgggca tggaaccaga
     6721 cctgccagcc ttgaccgaaa cggaggaatg ggaacggcgc gggcagcagc gcctgccgat
     6781 gcccgatgag ccgtgttttc tggacgatgg cgagccgttg gagccgccga cacgggtcac
     6841 gctgccgcgc cggtagcact tgggttgcgc agcaacccgt aagtgcgctg ttccagacta
     6901 tcggctgtag ccgcctcgcc gccctatacc ttgtctgcct ccccgcgttg cgtcgcggtg
     6961 catggagccg ggccacctcg acctgaatgg aagccggcgg cacctcgcta acggattcac
     7021 cgtttttatc aggctctggg aggcagaata aatgatcata tcgtcaatta ttacctccac
     7081 ggggagagcc tgagcaaact ggcctcaggc atttgagaag cacacggtca cactgcttcc
     7141 ggtagtcaat aaaccggtaa accagcaata gacataagcg gctatttaac gaccctgccc
     7201 tgaaccgacg accgggtcga atttgctttc gaatttctgc cattcatccg cttattatca
     7261 cttattcagg cgtagcacca ggcgtttaag tcgaccaata actctacggg gtctgacgct
     7321 cagtggaacg aa
//
