LOCUS       40924_26306        7711 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Integration expression vector pJM253, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7711)
  AUTHORS   Meisner J, Goldberg JB.
  TITLE     Escherichia coli rhaSR-PrhaBAD inducible promoter system allows 
            tightly controlled gene expression over a wide range in Pseudomonas 
            aeruginosa
  JOURNAL   Appl. Environ. Microbiol. (2016) In press
  PUBMED    27613678
REFERENCE   2  (bases 1 to 7711)
  AUTHORS   Meisner J.
  TITLE     Direct Submission
  JOURNAL   Submitted (26-AUG-2016) Biochemistry, Emory University School of 
            Medicine, 1510 Clifton Road NE, Rollins Research Center, room G223, 
            Atlanta, GA 30322, USA
REFERENCE   3  (bases 1 to 7711)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7711)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Appl. 
            Environ. Microbiol. (2016) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (26-AUG-2016) Biochemistry, Emory University School of Medicine, 
            1510 Clifton Road NE, Rollins Research Center, room G223, Atlanta, 
            GA 30322, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..7711
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        827..845
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             complement(968..1813)
                     /codon_start=1
                     /label=rhaR
                     /note="transcriptional activator of rhaR and rhaS"
                     /translation="VAHQLKLLKDDFFASDQQAVAVADRYPQDVFAEHTHDFCELVIVW
                     RGNGLHVLNDRPYRITRGDLFYIHADDKHSYASVNDLVLQNIIYCPERLKLNLDWQGAI
                     PGFNASAGQPHWRLGSMGMAQARQVIGQLEHESSQHVPFANEMAELLFGQLVMLLNRHR
                     YTSDSLPPTSSETLLDKLITRLAASLKSPFALDKFCDEASCSERVLRQQFRQQTGMTIN
                     QYLRQVRVCHAQYLLQHSRLLISDISTECGFEDSNYFSVVFTRETGMTPSQWRHLNSQK
                     D"
     CDS             complement(1890..2723)
                     /codon_start=1
                     /label=rhaS
                     /note="positive regulator of the rhaB promoter"
                     /translation="MTVLHSVDFFPSGNASVAIEPRLPQADFPEHHHDFHEIVIVEHGT
                     GIHVFNGQPYTITGGTVCFVRDHDRHLYEHTDNLCLTNVLYRSPDRFQFLAGLNQLLPQ
                     ELDGQYPSHWRVNHSVLQQVRQLVAQMEQQEGENDLPSTASREILFMQLLLLLRKSSLQ
                     ENLENSASRLNLLLAWLEDHFADEVNWDAVADQFSLSLRTLHRQLKQQTGLTPQRYLNR
                     LRLMKARHLLRHSEASVTDIAYRCGFSDSNHFSTLFRREFNWSPRDIRQGRDGFLQ"
     promoter        2869..2987
                     /label=rhaB promoter
                     /note="promoter of the E. coli rhaBAD operon, conferring
                     tight induction with L-rhamnose and repression with 
                     D-glucose in the presence of RhaR and RhaS (Giacalone et 
                     al., 2006)"
     primer_bind     complement(3033..3049)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(3075..3093)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    complement(3267..3314)
                     /label=FRT
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     oriT            complement(3521..3629)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     rep_origin      complement(5054..5642)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6467..7654)
                     /codon_start=1
                     /label=TcR
                     /note="tetracycline efflux protein"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     promoter        complement(join(7702..7711,1..19))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
ORIGIN
        1 atcgatgata agctgtcaaa catgagaatt aattcatgtt ctttcctgcg ttatcccctg
       61 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
      121 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc
      181 ctctccccgc gcgttggccg attcattaat gcagcgaatt agcttcaaaa gcgctctgaa
      241 gttcctatac tttctagaga ataggaactt cggaatagga acttcaagat cccctgattc
      301 cctttgtcaa cagcaatgga ttttcaatgg gtcaacatga aatccggctg ttgccctggg
      361 acggcaggtt tgacggcagg cttccaggct ctctcgcaca cgctgggaac actctgggaa
      421 caaacgcgag tgaaaatgga cagtagggcc caggaataat gcggcctcca gcgaggttgg
      481 ggcgtttcat atgagttcga atctcaccgc ctccgccata taacctgctg ttttcattga
      541 tgtttccatt cgacccactc tcaggagtga acgatttggg aacaaactgg gaacgaggcg
      601 cagaaaaaag gggccccgca gggccccttt ttcgttgctt caggaaagtt tcagcgcctg
      661 attcagcaga cctacgatgt cggggccgtc cttgctgaat tagctttatg cttgtaaacc
      721 gttttgtgaa aaaattttta aaataaaaaa ggggacctct agggtcccca attaattagt
      781 aatataatct attaaaggtc attcaaaagg tcatccaccg gcgcgcaatt aaccctcact
      841 aaagggaaca aaagctggag ctcaaagagt ggaacaatgc aggacgccgt ccggtaacgg
      901 tgttgagcct cggttgtgtg gtgattattg tcgccgctaa catcgtcggc atcggcatgg
      961 cgaattaatc tttctgcgaa ttgagatgac gccactggct gggcgtcatc ccggtttccc
     1021 gggtaaacac caccgaaaaa tagttactat cttcaaagcc acattcggtc gaaatatcac
     1081 tgattaacag gcggctatgc tggagaagat attgcgcatg acacactctg acctgtcgca
     1141 gatattgatt gatggtcatt ccagtctgct ggcgaaattg ctgacgcaaa acgcgctcac
     1201 tgcacgatgc ctcatcacaa aatttatcca gcgcaaaggg acttttcagg ctagccgcca
     1261 gccgggtaat cagcttatcc agcaacgttt cgctggatgt tggcggcaac gaatcactgg
     1321 tgtaacgatg gcgattcagc aacatcacca actgcccgaa cagcaactca gccatttcgt
     1381 tagcaaacgg cacatgctga ctactttcat gctcaagctg accgataacc tgccgcgcct
     1441 gcgccatccc catgctacct aagcgccagt gtggttgccc tgcgctggcg ttaaatcccg
     1501 gaatcgcccc ctgccagtca agattcagct tcagacgctc cgggcaataa ataatattct
     1561 gcaaaaccag atcgttaacg gaagcgtagg agtgtttatc gtcagcatga atgtaaaaga
     1621 gatcgccacg ggtaatgcga taagggcgat cgttgagtac atgcaggcca ttaccgcgcc
     1681 agacaatcac cagctcacaa aaatcatgtg tatgttcagc aaagacatct tgcggataac
     1741 ggtcagccac agcgactgcc tgctggtcgc tggcaaaaaa atcatctttg agaagtttta
     1801 actgatgcgc caccgtggct acctcggcca gagaacgaag ttgattattc gcaatatggc
     1861 gtacaaatac gttgagaaga ttcgcgttat tgcagaaagc catcccgtcc ctggcgaata
     1921 tcacgcggtg accagttaaa ctctcggcga aaaagcgtcg aaaagtggtt actgtcgctg
     1981 aatccacagc gataggcgat gtcagtaacg ctggcctcgc tgtggcgtag cagatgtcgg
     2041 gctttcatca gtcgcaggcg gttcaggtat cgctgaggcg tcagtcccgt ttgctgctta
     2101 agctgccgat gtagcgtacg cagtgaaaga gaaaattgat ccgccacggc atcccaattc
     2161 acctcatcgg caaaatggtc ctccagccag gccagaagca agttgagacg tgatgcgctg
     2221 ttttccaggt tctcctgcaa actgctttta cgcagcaaga gcagtaattg cataaacaag
     2281 atctcgcgac tggcggtcga gggtaaatca ttttcccctt cctgctgttc catctgtgca
     2341 accagctgtc gcacctgctg caatacgctg tggttaacgc gccagtgaga cggatactgc
     2401 ccatccagct cttgtggcag caactgattc agcccggcga gaaactgaaa tcgatccggc
     2461 gagcgataca gcacattggt cagacacaga ttatcggtat gttcatacag atgccgatca
     2521 tgatcgcgta cgaaacagac cgtgccaccg gtgatggtat agggctgccc attaaacaca
     2581 tgaatacccg tgccatgttc gacaatcaca atttcatgaa aatcatgatg atgttcagga
     2641 aaatccgcct gcgggagccg gggttctatc gccacggacg cgttaccaga cggaaaaaaa
     2701 tccacactat gtaatacggt catactggcc tcctgatgtc gtcaacacgg cgaaatagta
     2761 atcacgaggt caggttctta ccttaaattt tcgacggaaa accacgtaaa aaacgtcgat
     2821 ttttcaagat acagcgtgaa ttttcaggaa atgcggtgag catcacatca ccacaattca
     2881 gcaaattgtg aacatcatca cgttcatctt tccctggttg ccaatggccc attttcctgt
     2941 cagtaacgag aaggtcgcga attcaggcgc tttttagact ggtcgtaatg aaattcaact
     3001 agtgctctgc aggaattcct cgagaagctt atcgataccg tcgacctcga gggggggccc
     3061 ggtacccaat tcgccctata gtgagtcgta ttacgcgcgc cggtggatga ccttttgaat
     3121 gacctttaat agattatatt actaattaat tggggaccct agaggtcccc ttttttattt
     3181 taaaaatttt ttcacaaaac ggtttacaag cataaagcta attccattga gtaagttttt
     3241 aagcacatca gcttcaaaag cgctctgaag ttcctatact ttctagagaa taggaacttc
     3301 ggaataggaa cttcaagatc cccaattcgc tgcattaatg aatcggccaa cgcgcgggga
     3361 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     3421 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     3481 aatcagggga taacgcagga aagaacatgc tagagcttat cggccagcct cgcagagcag
     3541 gattcccgtt gagcaccgcc aggtgcgaat aagggacagt gaagaaggaa cacccgctcg
     3601 cgggtgggcc tacttcacct atcctgcccg gctgacgccg ttggatacac caaggaaagt
     3661 ctacacgaac cctttggcaa aatcctgtat atcgtgcgaa aaaggatgga tataccgaaa
     3721 aaatcgctat aatgaccccg aagcagggtt atgcagcgga aagtatacct taaccttttt
     3781 cgttgcttca ggaaagtttc agcgcctgat tcagcagacc tacgatgtcg gggccgtcct
     3841 tgctgatcca cttggcgtaa tgcttgaaga tcatcgccgt cgaggtgtgc cccatctggt
     3901 ccgcgatcca ttcgggagtg gcgatgccgc tgctgagcat ctggctggcg aaggtgtggc
     3961 ggcactgatt gggaccacgc gagcggacgc ccgcattcct gaggtggcca tgccaccagc
     4021 cgttgcgcag cgtatccgac gtggagtacg ccgccccgct agcactgttg tggaacacga
     4081 agcgtacccg ttgaggcttt ctggtccggt tgtcgcggtc tatcacctcg atctgtacgg
     4141 gcggaaggcg tctggttagc ttggcctgct gctggagtgc gcgcagcgcc ggggccagta
     4201 gctggacctt cctcgtcgat cggcgagtct tggtgacctt gtactggctg cgtacacgag
     4261 cgcggcggaa aaccacggtc cccgtgtcca aatcgacatc ttcccaggcc agagctatgg
     4321 cctcgctgac gcgggggccg ctccagatca tgaactccgc caggttcagc tcaccgatgc
     4381 gcgcggtttc agtaccgagg atgaggtcga tttcctcacg agtgaatggg tctgggtcat
     4441 ccgcatccgg aagagtgatc acgatgccgt ctgttgggtc gtgcgcgctc cggttccgag
     4501 ttcgatacag ccggaagatc tgccgcaggt tgctgacgat ctctcgcacc gtcttgttgt
     4561 gcagcttggg catgagcgtg ttctgcaccc agtcttgaat atccaggtgg tcgatgctgt
     4621 cggcctggct gtcaccccag cgaggtcgga tgtgggtttc cacgcggctg gtgtagccgc
     4681 ggaagccgct ggcggcaatc tggttgcgtt tgatgtcgag ccagaggtcg atgtaatggc
     4741 ccaaggtgtt gcttttcacc cgcggtgagt ccgggaagtg tcggctgtag ctgaagactc
     4801 cttgcttgat ctcgtagttg atgatgccgg ccagccgttc ggcattggct atattctcgg
     4861 gcgttgcgtc ccctggtatg gactccctac acagctcgcc ctggtagcgg aagtagatcc
     4921 tgatccgttt tccgcgtacc tcgactccgt ctgccatctg gttcccctga tccacaccaa
     4981 gcgttcaagc tggcatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     5041 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     5101 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     5161 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     5221 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     5281 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     5341 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     5401 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     5461 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     5521 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     5581 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     5641 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     5701 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     5761 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     5821 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     5881 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     5941 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     6001 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     6061 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     6121 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     6181 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     6241 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     6301 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     6361 gtgagtcaag ggttggtttg cgcattcaca gttctccgca agaattgatt ggctccaatt
     6421 cttggagtgg tgaatccgtt agcgaggtgc cgccggcttc cattcaggtc gaggtggccc
     6481 ggctccatgc accgcgacgc aacgcgggga ggcagacaag gtatagggcg gcgcctacaa
     6541 tccatgccaa cccgttccat gtgctcgccg aggcggcata aatcgccgtg acgatcagcg
     6601 gtccagtgat cgaagttagg ctggtaagag ccgcgagcga tccttgaagc tgtccctgat
     6661 ggtcgtcatc tacctgcctg gacagcatgg cctgcaacgc gggcatcccg atgccgccgg
     6721 aagcgagaag aatcataatg gggaaggcca tccagcctcg cgtcgcgaac gccagcaaga
     6781 cgtagcccag cgcgtcggcc gccatgccgg cgataatggc ctgcttctcg ccgaaacgtt
     6841 tggtggcggg accagtgacg aaggcttgag cgagggcgtg caagattccg aataccgcaa
     6901 gcgacaggcc gatcatcgtc gcgctccagc gaaagcggtc ctcgccgaaa atgacccaga
     6961 gcgctgccgg cacctgtcct acgagttgca tgataaagaa gacagtcata agtgcggcga
     7021 cgatagtcat gccccgcgcc caccggaagg agctgactgg gttgaaggct ctcaagggca
     7081 tcggtcggcg ctctccctta tgcgactcct gcattaggaa gcagcccagt agtaggttga
     7141 ggccgttgag caccgccgcc gcaaggaatg gtgcatgtaa ggagatggcg cccaacagtc
     7201 ccccggccac ggggcctgcc accataccca cgccgaaaca agcgctcatg agcccgaagt
     7261 ggcgagcccg atcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg
     7321 tggcgccggt gatgccggcc acgatgcgtc cggcgtagag aatccacagg acgggtgtgg
     7381 tcgccatgat cgcgtagtcg atagtggctc caagtagcga agcgagcagg actgggcggc
     7441 ggccaaagcg gtcggacagt gctccgagaa cgggtgcgca tagaaattgc atcaacgcat
     7501 atagcgctag cagcacgcca tagtgactgg cgatgctgtc ggaatggaca atatcccgca
     7561 agaggcccgg cagtaccggc ataaccaagc ctatgcctac agcatccagg gtgacggtgc
     7621 cgaggatgac gatgagcgca ttgttagatt tcatacacgg tgcctgactg cgttagcaat
     7681 ttaactgtga taaactaccg cattaaagct a
//
