LOCUS       V002117                10469 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V002117
VERSION     V002117
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 10469)
  AUTHORS   van Rossum T, Muras A, Baur MJ, Creutzburg SC, van der Oost J,
            Kengen SW.
  TITLE     A growth- and bioluminescence-based bioreporter for the in vivo
            detection of novel biocatalysts
  JOURNAL   Microb Biotechnol (2017) In press
   PUBMED   28393499
REFERENCE   2  (bases 1 to 10469)
  AUTHORS   van Rossum T, Muras A, Baur MJ, Creutzburg SC, van der Oost J,
            Kengen SW.
  TITLE     Direct Submission
  JOURNAL   Submitted (04-AUG-2016) Laboratory of Microbiology, Wageningen
            University and Research, Stippeneng 4, Wageningen 6708 WE, The
            Netherlands
REFERENCE   3  (bases 1 to 10469)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 10469)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Microb
            Biotechnol (2017) In press"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (04-AUG-2016) Laboratory of Microbiology, Wageningen University and
            Research, Stippeneng 4, Wageningen 6708 WE, The Netherlands"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..10469
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        10..87
                     /label="lacIq promoter"
                     /note="In the lacIq allele, a single base change in the
                     promoter boosts expression of the lacI gene about 10-fold."
     regulatory      95..100
                     /label="RBS"
                     /note="RBS"
                     /regulatory_class="ribosome_binding_site"
     CDS             107..982
                     /label="araC"
                     /note="L-arabinose regulatory protein"
     CDS             complement(995..2083)
                     /gene="leuB"
                     /label="3-isopropylmalate dehydrogenase"
                     /note="3-isopropylmalate dehydrogenase from Escherichia
                     coli (strain K12). Accession#: P30125"
     regulatory      complement(2090..2095)
                     /label="RBS"
                     /note="RBS"
                     /regulatory_class="ribosome_binding_site"
     regulatory      complement(2104..2414)
                     /label="PBAD-adapt"
                     /note="PBAD-adapt"
                     /regulatory_class="promoter"
     rep_origin      complement(2565..3153)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     regulatory      3235..3545
                     /label="PBAD-adapt"
                     /note="PBAD-adapt"
                     /regulatory_class="promoter"
     regulatory      3552..3557
                     /label="RBS"
                     /note="RBS"
                     /regulatory_class="ribosome_binding_site"
     CDS             3564..5003
                     /label="LuxC"
                     /note="LuxC fatty acid reductase"
     CDS             5019..5939
                     /label="LuxD"
                     /note="LuxD acyltransferase"
     CDS             5991..7070
                     /label="LuxA"
                     /note="LuxA luciferase subunit"
     CDS             7088..8068
                     /label="LuxB"
                     /note="LuxB luciferase subunit"
     CDS             8250..9359
                     /label="LuxE"
                     /note="LuxE"
     terminator      9382..9468
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      complement(9494..9588)
                     /label="lambda t0 terminator"
                     /note="transcription terminator from phage lambda"
     CDS             complement(9612..10268)
                     /label="CmR"
                     /note="chloramphenicol acetyltransferase"
     promoter        complement(10269..10371)
                     /label="cat promoter"
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
ORIGIN
        1 ctcgaggttg acaccatcga atggtgcaaa acctttcgcg gtatggcatg atagcgcccg
       61 gaagagagtc aattcagggt ggtgaatcgg tccgaggagg atacgtatgg ctgaagcgca
      121 aaatgatccc ctgctgccgg gatactcgtt taatgcccat ctggtggcgg gtttaacgcc
      181 gattgaggcc aacggttatc tcgatttttt tatcgaccga ccgctgggaa tgaaaggtta
      241 tattctcaat ctcaccattc gcggtcaggg ggtggtgaaa aatcagggac gagaatttgt
      301 ttgccgaccg ggtgatattt tgctgttccc gccaggagag attcatcact acggtcgtca
      361 tccggaggct cgcgaatggt atcaccagtg ggtttacttt cgtccgcgcg cctactggca
      421 tgaatggctt aactggccgt caatatttgc caatacgggg ttctttcgcc cggatgaagc
      481 gcaccagccg catttcagcg acctgtttgg gcaaatcatt aacgccgggc aaggggaagg
      541 gcgctattcg gagctgctgg cgataaatct gcttgagcaa ttgttactgc ggcgcatgga
      601 agcgattaac gagtcgctcc atccaccgat ggataatcgg gtacgcgagg cttgtcagta
      661 catcagcgat cacctggcag acagcaattt tgatatcgcc agcgtcgcac agcatgtttg
      721 cttgtcgccg tcgcgtctgt cacatctttt ccgccagcag ttagggatta gcgtcttaag
      781 ctggcgcgag gaccaacgta tcagccaggc gaagctgctt ttgagcacca cccggatgcc
      841 tatcgccacc gtcggtcgca atgttggttt tgacgatcaa ctctatttct cgcgggtatt
      901 taaaaaatgc accggggcca gcccgagcga gttccgtgcc ggttgtgaag aaaaagtgaa
      961 tgatgtagcc gtcaagttgt cataacgatc gttacacccc ttctgctaca tagcgggcaa
     1021 tgatatcgcc catttcatcg gtactaacgg cggcagcgcc acgggctaaa tccccggtgc
     1081 gaatgccttc ttctaatgcg cggttaatgg cgcgttcaat ggcgcaagcc gcatcatcgg
     1141 catccaggct gtaacgcagc agcagtgcca gcgaaaggat ttgtgcaatc gggttggcga
     1201 tgtttttgcc tgcgatatct ggtgccgagc cgcccgccgg ttcatacagt ccaaaacctt
     1261 gctcgttcag gctggcggaa ggcaacatcc ccatcgagcc agtgatcatt gcgcactcgt
     1321 cagacagaat gtcgccaaac aggttggagc acagcagaac gtcaaactgt gatggatctt
     1381 taatcagctg catggtggcg ttgtcgatgt acatatgcgc cagttcgaca tccgggtatt
     1441 ccgtggcgat ctcgttaacg atctcccgcc ataaaataga ggattgcagc acgttggctt
     1501 tatcgatact cgtcactttg tggcgacgct tgcgagcaga ttcaaacgcg atgcgggcga
     1561 tacgttcgat ctcaaaacgg tgatacacct cggtatcaaa ggctttttca tattgtccgc
     1621 taccttcgcg gccttttggc tgaccgaaat agatgccgcc ggtcagttcg cgcacacaca
     1681 ggatgtcgaa gccgtttgcg gcaatgtctg cacgcagcgg acagaatgct tccagcccct
     1741 gatacagttt tgccgggcgc aggttgctga ataatttgaa gtgcttacgc agaggcagca
     1801 gcgcgccgcg ttctggttgc tggtctggtg gtaaatgttc ccacttcggg ccgcctaccg
     1861 agccaaacag cacggcatcg gcttgctcac aaccttcaac cgtcgcaggc ggcagtggtt
     1921 gcccgtggtt atcaatggct gcgccgccta catcgtaatg gctggtggtg atgcgcatcg
     1981 caaagcggtt gcgcacggca tccagcactt tcagcgcctg ggtcatcact tccggaccaa
     2041 taccgtcccc cggcaatacg gcaatatggt aattcttcga catacgtatc ctcctcgccg
     2101 gcgcccaaaa aaacgggtat ggagaaacag tagagagttg cgataaaaag cgtcaggtag
     2161 gatccgctaa tcttatggat aaaaatgcta tgcgagctag cttccatctc gacttaggtt
     2221 aatcaatgtg gacttttctg ccgtgattat agacactttt gttacgcgtt tttgtcatgg
     2281 ctttggtccc gctttgttac agaatgcttt taataagcgg ggttaccggt ttggttagcg
     2341 agaagagcca gtaaaagacg cagtgacggc aatgtctgat gcaatatgga caattggttt
     2401 cttctctgaa tggccctagg cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     2461 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     2521 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     2581 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     2641 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     2701 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     2761 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     2821 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     2881 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     2941 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     3001 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     3061 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     3121 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     3181 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatggg taccgccatt
     3241 cagagaagaa accaattgtc catattgcat cagacattgc cgtcactgcg tcttttactg
     3301 gctcttctcg ctaaccaaac cggtaacccc gcttattaaa agcattctgt aacaaagcgg
     3361 gaccaaagcc atgacaaaaa cgcgtaacaa aagtgtctat aatcacggca gaaaagtcca
     3421 cattgattaa cctaagtcga gatggaactg cagtcgcata gcatttttat ccataagatt
     3481 agcggatcct acctgacgct ttttatcgca actctctact gtttctccat acccgttttt
     3541 ttggggtcga caggaggata cgtatgacta aaaaaatttc attcattatt aacggccagg
     3601 ttgaaatctt tcccgaaggt gatgatttag tgcaatccat taattttggt gataatagtg
     3661 tttacctgcc aatattgaat gactctcatg taaaaaacat tattgattgt aatggaaata
     3721 acgaattacg gttgcataac attgtcaatt ttctctatac ggtagggcaa agatggaaaa
     3781 atgaagaata ctcaagacgc aggacataca ttcgtgactt aaaaaaatat atgggatatt
     3841 cagaagaaat ggctaagcta gaggccaatt ggatatctat gattttatgt tctaaaggcg
     3901 gcctttatga tgttgtagaa aatgaacttg gttctcgcca tatcatggat gaatggctac
     3961 ctcaggatga aagttatgtt cgggcttttc cgaaaggtaa atctgtacat ctgttggcag
     4021 gtaatgttcc attatctggg atcatgtcta tattacgcgc aattttaact aagaatcagt
     4081 gtattataaa aacatcgtca accgatcctt ttaccgctaa tgcattagcg ttaagtttta
     4141 ttgatgtaga ccctaatcat ccgataacgc gctctttatc tgttatatat tggccccacc
     4201 aaggtgatac atcactcgca aaagaaatta tgcaacatgc ggatgttatt gtcgcttggg
     4261 gagggccaga tgcgattaat tgggcggtag agcatgcgcc atcttatgct gatgtgatta
     4321 aatttggttc taaaaagagt ctttgcatta tcgataatcc tgttgatttg acgtccgcag
     4381 cgacaggtgc ggctcatgat gtttgttttt acgatcagcg agcttgtttt tctgcccaaa
     4441 acatatatta catgggaaat cattatgagg aatttaagtt agcgttgata gaaaaactta
     4501 atctatatgc gcatatatta ccgaatgcca aaaaagattt tgatgaaaag gcggcctatt
     4561 ctttagttca aaaagaaagc ttgtttgctg gattaaaagt agaggtggat attcatcaac
     4621 gttggatgat tattgagtca aatgcaggtg tggaatttaa tcaaccactt ggcagatgtg
     4681 tgtaccttca tcacgtcgat aatattgagc aaatattgcc ttatgttcaa aaaaataaga
     4741 cgcaaaccat atctattttt ccttgggagt catcatttaa atatcgagat gcgttagcat
     4801 taaaaggtgc ggaaaggatt gtagaagcag gaatgaataa catatttcga gttggtggat
     4861 ctcatgacgg aatgagaccg ttgcaacgat tagtgacata tatttctcat gaaaggccat
     4921 ctaactatac ggctaaggat gttgcggttg aaatagaaca gactcgattc ctggaagaag
     4981 ataagttcct tgtatttgtc ccataatagg taaaaagtat ggaaaatgaa tcaaaatata
     5041 aaaccatcga ccacgttatt tgtgttgaag gaaataaaaa aattcatgtt tgggaaacgc
     5101 tgccagaaga aaacagccca aagagaaaga atgccattat tattgcgtct ggttttgccc
     5161 gcaggatgga tcattttgct ggtctggcgg aatatttatc gcggaatgga tttcatgtga
     5221 tccgctatga ttcgcttcac cacgttggat tgagttcagg gacaattgat gaatttacaa
     5281 tgtctatagg aaagcagagc ttgttagcag tggttgattg gttaactaca cgaaaaataa
     5341 ataacttcgg tatgttggct tcaagcttat ctgcgcggat agcttatgca agcctatctg
     5401 aaatcaatgc ttcgttttta atcaccgcag tcggtgttgt taacttaaga tattctcttg
     5461 aaagagcttt agggtttgat tatctcagtc tacccattaa tgaattgccg aataatctag
     5521 attttgaagg ccataaattg ggtgctgaag tctttgcgag agattgtctt gattttggtt
     5581 gggaagattt agcttctaca attaataaca tgatgtatct tgatataccg tttattgctt
     5641 ttactgcaaa taacgataat tgggtcaagc aagatgaagt tatcacattg ttatcaaata
     5701 ttcgtagtaa tcgatgcaag atatattctt tgttaggaag ttcgcatgac ttgagtgaaa
     5761 atttagtggt cctgcgcaat ttttatcaat cggttacgaa agccgctatc gcgatggata
     5821 atgatcatct ggatattgat gttgatatta ctgaaccgtc atttgaacat ttaactattg
     5881 cgacagtcaa tgaacgccga atgagaattg agattgaaaa tcaagcaatt tctctgtctt
     5941 aaaatctatt gagatattct atcactcaaa tagcaatata aggactctct atgaaatttg
     6001 gaaacttttt gcttacatac caacctcccc aattttctca aacagaggta atgaaacgtt
     6061 tggttaaatt aggtcgcatc tctgaggagt gtggttttga taccgtatgg ttactggagc
     6121 atcatttcac ggagtttggt ttgcttggta acccttatgt cgctgctgca tatttacttg
     6181 gcgcgactaa aaaattgaat gtaggaactg ccgctattgt tcttcccaca gcccatccag
     6241 tacgccaact tgaagatgtg aatttattgg atcaaatgtc aaaaggacga tttcggtttg
     6301 gtatttgccg agggctttac aacaaggact ttcgcgtatt cggcacagat atgaataaca
     6361 gtcgcgcctt agcggaatgc tggtacgggc tgataaagaa tggcatgaca gagggatata
     6421 tggaagctga taatgaacat atcaagttcc ataaggtaaa agtaaacccc gcggcgtata
     6481 gcagaggtgg cgcaccggtt tatgtggtgg ctgaatcagc ttcgacgact gagtgggctg
     6541 ctcaatttgg cctaccgatg atattaagtt ggattataaa tactaacgaa aagaaagcac
     6601 aacttgagct ttataatgaa gtggctcaag aatatgggca cgatattcat aatatcgacc
     6661 attgcttatc atatataaca tctgtagatc atgactcaat taaagcgaaa gagatttgcc
     6721 ggaaatttct ggggcattgg tatgattctt atgtgaatgc tacgactatt tttgatgatt
     6781 cagaccaaac aagaggttat gatttcaata aagggcagtg gcgtgacttt gtattaaaag
     6841 gacataaaga tactaatcgc cgtattgatt acagttacga aatcaatccc gtgggaacgc
     6901 cgcaggaatg tattgacata attcaaaaag acattgatgc tacaggaata tcaaatattt
     6961 gttgtggatt tgaagctaat ggaacagtag acgaaattat tgcttccatg aagctcttcc
     7021 agtctgatgt catgccattt cttaaagaaa aacaacgttc gctattatat tagctaagga
     7081 gaaagaaatg aaatttggat tgttcttcct taacttcatc aattcaacaa ctgttcaaga
     7141 acaaagtata gttcgcatgc aggaaataac ggagtatgtt gataagttga attttgaaca
     7201 gattttagtg tatgaaaatc atttttcaga taatggtgtt gtcggcgctc ctctgactgt
     7261 ttctggtttt ctgctcggtt taacagagaa aattaaaatt ggttcattaa atcacatcat
     7321 tacaactcat catcctgtcc gcatagcgga ggaagcttgc ttattggatc agttaagtga
     7381 agggagattt attttagggt ttagtgattg cgaaaaaaaa gatgaaatgc atttttttaa
     7441 tcgcccggtt gaatatcaac agcaactatt tgaagagtgt tatgaaatca ttaacgatgc
     7501 tttaacaaca ggctattgta atccagataa cgatttttat agcttcccta aaatatctgt
     7561 aaatccccat gcttatacgc caggcggacc tcggaaatat gtaacagcaa ccagtcatca
     7621 tattgttgag tgggcggcca aaaaaggtat tcctctcatc tttaagtggg atgattctaa
     7681 tgatgttaga tatgaatatg ctgaaagata taaagccgtt gcggataaat atgacgttga
     7741 cctatcagag atagaccatc agttaatgat attagttaac tataacgaag atagtaataa
     7801 agctaaacaa gagacgcgtg catttattag tgattatgtt cttgaaatgc accctaatga
     7861 aaatttcgaa aataaacttg aagaaataat tgcagaaaac gctgtcggaa attatacgga
     7921 gtgtataact gcggctaagt tggcaattga aaagtgtggt gcgaaaagtg tattgctgtc
     7981 ctttgaacca atgaatgatt tgatgagcca aaaaaatgta atcaatattg ttgatgataa
     8041 tattaagaag taccacatgg aatataccta atagatttcg agttgcagcg aggcggcaag
     8101 tgaacgaatc cccaggagca tagataacta tgtgactggg gtgagtgaaa gcagccaaca
     8161 aagcagcagc ttgaaagatg aagggtataa aagagtatga cagcagtgct gccatacttt
     8221 ctaatattat cttgaggagt aaaacaggta tgacttcata tgttgataaa caagaaatta
     8281 cagcaagctc agaaattgat gatttgattt tttcgagcga tccattagtg tggtcttacg
     8341 acgagcagga aaaaatcaga aagaaacttg tgcttgatgc atttcgtaat cattataaac
     8401 attgtcgaga atatcgtcac tactgtcagg cacacaaagt agatgacaat attacggaaa
     8461 ttgatgacat acctgtattc ccaacatcgg tttttaagtt tactcgctta ttaacttctc
     8521 aggaaaacga gattgaaagt tggtttacca gtagcggcac gaatggttta aaaagtcagg
     8581 tggcgcgtga cagattaagt attgagagac tcttaggctc tgtgagttat ggcatgaaat
     8641 atgttggtag ttggtttgat catcaaatag aattagtcaa tttgggacca gatagattta
     8701 atgctcataa tatttggttt aaatatgtta tgagtttggt ggaattgtta tatcctacga
     8761 catttaccgt aacagaagaa cgaatagatt ttgttaaaac attgaatagt cttgaacgaa
     8821 taaaaaatca agggaaagat ctttgtctta ttggttcgcc atactttatt tatttactct
     8881 gccattatat gaaagataaa aaaatctcat tttctggaga taaaagcctt tatatcataa
     8941 ccggaggcgg ctggaaaagt tacgaaaaag aatctctgaa acgtgatgat ttcaatcatc
     9001 ttttatttga tactttcaat ctcagtgata ttagtcagat ccgagatata tttaatcaag
     9061 ttgaactcaa cacttgtttc tttgaggatg aaatgcagcg taaacatgtt ccgccgtggg
     9121 tatatgcgcg agcgcttgat cctgaaacgt tgaaacctgt acctgatgga acgccggggt
     9181 tgatgagtta tatggatgcg tcagcaacca gttatccagc atttattgtt accgatgatg
     9241 tcgggataat tagcagagaa tatggtaagt atcccggcgt gctcgttgaa attttacgtc
     9301 gcgtcaatac gaggacgcag aaagggtgtg ctttaagctt aaccgaagcg tttgatagtt
     9361 gagcggccgc tctagaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt
     9421 cgttttatct gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc gccgccctag
     9481 acctagtgct tggattctca ccaataaaaa acgcccggcg gcaaccgagc gttctgaaca
     9541 aatccagatg gagttctgag gtcattactg gatctatcaa caggagtcca agcgagctcg
     9601 atatcaaatt acgccccgcc ctgccactca tcgcagtact gttgtaattc attaagcatt
     9661 ctgccgacat ggaagccatc acagacggca tgatgaacct gaatcgccag cggcatcagc
     9721 accttgtcgc cttgcgtata atatttgccc atggtgaaaa cgggggcgaa gaagttgtcc
     9781 atattggcca cgtttaaatc aaaactggtg aaactcaccc agggattggc tgagacgaaa
     9841 aacatattct caataaaccc tttagggaaa taggccaggt tttcaccgta acacgccaca
     9901 tcttgcgaat atatgtgtag aaactgccgg aaatcgtcgt ggtattcact ccagagcgat
     9961 gaaaacgttt cagtttgctc atggaaaacg gtgtaacaag ggtgaacact atcccatatc
    10021 accagctcac cgtctttcat tgccatacgg aattccggat gagcattcat caggcgggca
    10081 agaatgtgaa taaaggccgg ataaaacttg tgcttatttt tctttacggt ctttaaaaag
    10141 gccgtaatat ccagctgaac ggtctggtta taggtacatt gagcaactga ctgaaatgcc
    10201 tcaaaatgtt ctttacgatg ccattgggat atatcaacgg tggtatatcc agtgattttt
    10261 ttctccattt tagcttcctt agctcctgaa aatctcgata actcaaaaaa tacgcccggt
    10321 agtgatctta tttcattatg gtgaaagttg gaacctctta cgtgccgatc aacgtctcat
    10381 tttcgccaga tatcgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
    10441 ggcgtatcac gaggcccttt cgtcttcac
//
