LOCUS       V002115                10570 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V002115
VERSION     V002115
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 10570)
  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 10570)
  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 10570)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 10570)
  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..10570
                     /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      2521..3066
                     /label="p15A ori"
                     /note="Plasmids containing the medium-copy-number p15A
                     origin of replication can be propagated in E. coli cells
                     that contain a second plasmid with the ColE1 origin."
     regulatory      3336..3646
                     /label="PBAD-adapt"
                     /note="PBAD-adapt"
                     /regulatory_class="promoter"
     regulatory      3653..3658
                     /label="RBS"
                     /note="RBS"
                     /regulatory_class="ribosome_binding_site"
     CDS             3665..5104
                     /label="LuxC"
                     /note="LuxC fatty acid reductase"
     CDS             5120..6040
                     /label="LuxD"
                     /note="LuxD acyltransferase"
     CDS             6092..7171
                     /label="LuxA"
                     /note="LuxA luciferase subunit"
     CDS             7189..8169
                     /label="LuxB"
                     /note="LuxB luciferase subunit"
     CDS             8351..9460
                     /label="LuxE"
                     /note="LuxE"
     terminator      9483..9569
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      complement(9595..9689)
                     /label="lambda t0 terminator"
                     /note="transcription terminator from phage lambda"
     CDS             complement(9713..10369)
                     /label="CmR"
                     /note="chloramphenicol acetyltransferase"
     promoter        complement(10370..10472)
                     /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 acaacttata tcgtatgggg ctgacttcag gtgctacatt
     2461 tgaagagata aattgcactg aaatctagaa atattttatc tgattaataa gatgatcttc
     2521 ttgagatcgt tttggtctgc gcgtaatctc ttgctctgaa aacgaaaaaa ccgccttgca
     2581 gggcggtttt tcgaaggttc tctgagctac caactctttg aaccgaggta actggcttgg
     2641 aggagcgcag tcaccaaaac ttgtcctttc agtttagcct taaccggcgc atgacttcaa
     2701 gactaactcc tctaaatcaa ttaccagtgg ctgctgccag tggtgctttt gcatgtcttt
     2761 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcggac tgaacggggg
     2821 gttcgtgcat acagtccagc ttggagcgaa ctgcctaccc ggaactgagt gtcaggcgtg
     2881 gaatgagaca aacgcggcca taacagcgga atgacaccgg taaaccgaaa ggcaggaaca
     2941 ggagagcgca cgagggagcc gccaggggga aacgcctggt atctttatag tcctgtcggg
     3001 tttcgccacc actgatttga gcgtcagatt tcgtgatgct tgtcaggggg gcggagccta
     3061 tggaaaaacg gctttgccgc ggccctctca cttccctgtt aagtatcttc ctggcatctt
     3121 ccaggaaatc tccgccccgt tcgtaagcca tttccgctcg ccgcagtcga acgaccgagc
     3181 gtagcgagtc agtgagcgag gaagcggaat atatcctgta tcacatattc tgctgacgca
     3241 ccggtgcagc cttttttctc ctgccacatg aagcacttca ctgacaccct catcagtgcc
     3301 aacatagtaa gccagtatac actccgctag gtaccgccat tcagagaaga aaccaattgt
     3361 ccatattgca tcagacattg ccgtcactgc gtcttttact ggctcttctc gctaaccaaa
     3421 ccggtaaccc cgcttattaa aagcattctg taacaaagcg ggaccaaagc catgacaaaa
     3481 acgcgtaaca aaagtgtcta taatcacggc agaaaagtcc acattgatta acctaagtcg
     3541 agatggaact gcagtcgcat agcattttta tccataagat tagcggatcc tacctgacgc
     3601 tttttatcgc aactctctac tgtttctcca tacccgtttt tttggggtcg acaggaggat
     3661 acgtatgact aaaaaaattt cattcattat taacggccag gttgaaatct ttcccgaagg
     3721 tgatgattta gtgcaatcca ttaattttgg tgataatagt gtttacctgc caatattgaa
     3781 tgactctcat gtaaaaaaca ttattgattg taatggaaat aacgaattac ggttgcataa
     3841 cattgtcaat tttctctata cggtagggca aagatggaaa aatgaagaat actcaagacg
     3901 caggacatac attcgtgact taaaaaaata tatgggatat tcagaagaaa tggctaagct
     3961 agaggccaat tggatatcta tgattttatg ttctaaaggc ggcctttatg atgttgtaga
     4021 aaatgaactt ggttctcgcc atatcatgga tgaatggcta cctcaggatg aaagttatgt
     4081 tcgggctttt ccgaaaggta aatctgtaca tctgttggca ggtaatgttc cattatctgg
     4141 gatcatgtct atattacgcg caattttaac taagaatcag tgtattataa aaacatcgtc
     4201 aaccgatcct tttaccgcta atgcattagc gttaagtttt attgatgtag accctaatca
     4261 tccgataacg cgctctttat ctgttatata ttggccccac caaggtgata catcactcgc
     4321 aaaagaaatt atgcaacatg cggatgttat tgtcgcttgg ggagggccag atgcgattaa
     4381 ttgggcggta gagcatgcgc catcttatgc tgatgtgatt aaatttggtt ctaaaaagag
     4441 tctttgcatt atcgataatc ctgttgattt gacgtccgca gcgacaggtg cggctcatga
     4501 tgtttgtttt tacgatcagc gagcttgttt ttctgcccaa aacatatatt acatgggaaa
     4561 tcattatgag gaatttaagt tagcgttgat agaaaaactt aatctatatg cgcatatatt
     4621 accgaatgcc aaaaaagatt ttgatgaaaa ggcggcctat tctttagttc aaaaagaaag
     4681 cttgtttgct ggattaaaag tagaggtgga tattcatcaa cgttggatga ttattgagtc
     4741 aaatgcaggt gtggaattta atcaaccact tggcagatgt gtgtaccttc atcacgtcga
     4801 taatattgag caaatattgc cttatgttca aaaaaataag acgcaaacca tatctatttt
     4861 tccttgggag tcatcattta aatatcgaga tgcgttagca ttaaaaggtg cggaaaggat
     4921 tgtagaagca ggaatgaata acatatttcg agttggtgga tctcatgacg gaatgagacc
     4981 gttgcaacga ttagtgacat atatttctca tgaaaggcca tctaactata cggctaagga
     5041 tgttgcggtt gaaatagaac agactcgatt cctggaagaa gataagttcc ttgtatttgt
     5101 cccataatag gtaaaaagta tggaaaatga atcaaaatat aaaaccatcg accacgttat
     5161 ttgtgttgaa ggaaataaaa aaattcatgt ttgggaaacg ctgccagaag aaaacagccc
     5221 aaagagaaag aatgccatta ttattgcgtc tggttttgcc cgcaggatgg atcattttgc
     5281 tggtctggcg gaatatttat cgcggaatgg atttcatgtg atccgctatg attcgcttca
     5341 ccacgttgga ttgagttcag ggacaattga tgaatttaca atgtctatag gaaagcagag
     5401 cttgttagca gtggttgatt ggttaactac acgaaaaata aataacttcg gtatgttggc
     5461 ttcaagctta tctgcgcgga tagcttatgc aagcctatct gaaatcaatg cttcgttttt
     5521 aatcaccgca gtcggtgttg ttaacttaag atattctctt gaaagagctt tagggtttga
     5581 ttatctcagt ctacccatta atgaattgcc gaataatcta gattttgaag gccataaatt
     5641 gggtgctgaa gtctttgcga gagattgtct tgattttggt tgggaagatt tagcttctac
     5701 aattaataac atgatgtatc ttgatatacc gtttattgct tttactgcaa ataacgataa
     5761 ttgggtcaag caagatgaag ttatcacatt gttatcaaat attcgtagta atcgatgcaa
     5821 gatatattct ttgttaggaa gttcgcatga cttgagtgaa aatttagtgg tcctgcgcaa
     5881 tttttatcaa tcggttacga aagccgctat cgcgatggat aatgatcatc tggatattga
     5941 tgttgatatt actgaaccgt catttgaaca tttaactatt gcgacagtca atgaacgccg
     6001 aatgagaatt gagattgaaa atcaagcaat ttctctgtct taaaatctat tgagatattc
     6061 tatcactcaa atagcaatat aaggactctc tatgaaattt ggaaactttt tgcttacata
     6121 ccaacctccc caattttctc aaacagaggt aatgaaacgt ttggttaaat taggtcgcat
     6181 ctctgaggag tgtggttttg ataccgtatg gttactggag catcatttca cggagtttgg
     6241 tttgcttggt aacccttatg tcgctgctgc atatttactt ggcgcgacta aaaaattgaa
     6301 tgtaggaact gccgctattg ttcttcccac agcccatcca gtacgccaac ttgaagatgt
     6361 gaatttattg gatcaaatgt caaaaggacg atttcggttt ggtatttgcc gagggcttta
     6421 caacaaggac tttcgcgtat tcggcacaga tatgaataac agtcgcgcct tagcggaatg
     6481 ctggtacggg ctgataaaga atggcatgac agagggatat atggaagctg ataatgaaca
     6541 tatcaagttc cataaggtaa aagtaaaccc cgcggcgtat agcagaggtg gcgcaccggt
     6601 ttatgtggtg gctgaatcag cttcgacgac tgagtgggct gctcaatttg gcctaccgat
     6661 gatattaagt tggattataa atactaacga aaagaaagca caacttgagc tttataatga
     6721 agtggctcaa gaatatgggc acgatattca taatatcgac cattgcttat catatataac
     6781 atctgtagat catgactcaa ttaaagcgaa agagatttgc cggaaatttc tggggcattg
     6841 gtatgattct tatgtgaatg ctacgactat ttttgatgat tcagaccaaa caagaggtta
     6901 tgatttcaat aaagggcagt ggcgtgactt tgtattaaaa ggacataaag atactaatcg
     6961 ccgtattgat tacagttacg aaatcaatcc cgtgggaacg ccgcaggaat gtattgacat
     7021 aattcaaaaa gacattgatg ctacaggaat atcaaatatt tgttgtggat ttgaagctaa
     7081 tggaacagta gacgaaatta ttgcttccat gaagctcttc cagtctgatg tcatgccatt
     7141 tcttaaagaa aaacaacgtt cgctattata ttagctaagg agaaagaaat gaaatttgga
     7201 ttgttcttcc ttaacttcat caattcaaca actgttcaag aacaaagtat agttcgcatg
     7261 caggaaataa cggagtatgt tgataagttg aattttgaac agattttagt gtatgaaaat
     7321 catttttcag ataatggtgt tgtcggcgct cctctgactg tttctggttt tctgctcggt
     7381 ttaacagaga aaattaaaat tggttcatta aatcacatca ttacaactca tcatcctgtc
     7441 cgcatagcgg aggaagcttg cttattggat cagttaagtg aagggagatt tattttaggg
     7501 tttagtgatt gcgaaaaaaa agatgaaatg cattttttta atcgcccggt tgaatatcaa
     7561 cagcaactat ttgaagagtg ttatgaaatc attaacgatg ctttaacaac aggctattgt
     7621 aatccagata acgattttta tagcttccct aaaatatctg taaatcccca tgcttatacg
     7681 ccaggcggac ctcggaaata tgtaacagca accagtcatc atattgttga gtgggcggcc
     7741 aaaaaaggta ttcctctcat ctttaagtgg gatgattcta atgatgttag atatgaatat
     7801 gctgaaagat ataaagccgt tgcggataaa tatgacgttg acctatcaga gatagaccat
     7861 cagttaatga tattagttaa ctataacgaa gatagtaata aagctaaaca agagacgcgt
     7921 gcatttatta gtgattatgt tcttgaaatg caccctaatg aaaatttcga aaataaactt
     7981 gaagaaataa ttgcagaaaa cgctgtcgga aattatacgg agtgtataac tgcggctaag
     8041 ttggcaattg aaaagtgtgg tgcgaaaagt gtattgctgt cctttgaacc aatgaatgat
     8101 ttgatgagcc aaaaaaatgt aatcaatatt gttgatgata atattaagaa gtaccacatg
     8161 gaatatacct aatagatttc gagttgcagc gaggcggcaa gtgaacgaat ccccaggagc
     8221 atagataact atgtgactgg ggtgagtgaa agcagccaac aaagcagcag cttgaaagat
     8281 gaagggtata aaagagtatg acagcagtgc tgccatactt tctaatatta tcttgaggag
     8341 taaaacaggt atgacttcat atgttgataa acaagaaatt acagcaagct cagaaattga
     8401 tgatttgatt ttttcgagcg atccattagt gtggtcttac gacgagcagg aaaaaatcag
     8461 aaagaaactt gtgcttgatg catttcgtaa tcattataaa cattgtcgag aatatcgtca
     8521 ctactgtcag gcacacaaag tagatgacaa tattacggaa attgatgaca tacctgtatt
     8581 cccaacatcg gtttttaagt ttactcgctt attaacttct caggaaaacg agattgaaag
     8641 ttggtttacc agtagcggca cgaatggttt aaaaagtcag gtggcgcgtg acagattaag
     8701 tattgagaga ctcttaggct ctgtgagtta tggcatgaaa tatgttggta gttggtttga
     8761 tcatcaaata gaattagtca atttgggacc agatagattt aatgctcata atatttggtt
     8821 taaatatgtt atgagtttgg tggaattgtt atatcctacg acatttaccg taacagaaga
     8881 acgaatagat tttgttaaaa cattgaatag tcttgaacga ataaaaaatc aagggaaaga
     8941 tctttgtctt attggttcgc catactttat ttatttactc tgccattata tgaaagataa
     9001 aaaaatctca ttttctggag ataaaagcct ttatatcata accggaggcg gctggaaaag
     9061 ttacgaaaaa gaatctctga aacgtgatga tttcaatcat cttttatttg atactttcaa
     9121 tctcagtgat attagtcaga tccgagatat atttaatcaa gttgaactca acacttgttt
     9181 ctttgaggat gaaatgcagc gtaaacatgt tccgccgtgg gtatatgcgc gagcgcttga
     9241 tcctgaaacg ttgaaacctg tacctgatgg aacgccgggg ttgatgagtt atatggatgc
     9301 gtcagcaacc agttatccag catttattgt taccgatgat gtcgggataa ttagcagaga
     9361 atatggtaag tatcccggcg tgctcgttga aattttacgt cgcgtcaata cgaggacgca
     9421 gaaagggtgt gctttaagct taaccgaagc gtttgatagt tgagcggccg ctctagaggc
     9481 atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt
     9541 cggtgaacgc tctcctgagt aggacaaatc cgccgcccta gacctagtgc ttggattctc
     9601 accaataaaa aacgcccggc ggcaaccgag cgttctgaac aaatccagat ggagttctga
     9661 ggtcattact ggatctatca acaggagtcc aagcgagctc gatatcaaat tacgccccgc
     9721 cctgccactc atcgcagtac tgttgtaatt cattaagcat tctgccgaca tggaagccat
     9781 cacagacggc atgatgaacc tgaatcgcca gcggcatcag caccttgtcg ccttgcgtat
     9841 aatatttgcc catggtgaaa acgggggcga agaagttgtc catattggcc acgtttaaat
     9901 caaaactggt gaaactcacc cagggattgg ctgagacgaa aaacatattc tcaataaacc
     9961 ctttagggaa ataggccagg ttttcaccgt aacacgccac atcttgcgaa tatatgtgta
    10021 gaaactgccg gaaatcgtcg tggtattcac tccagagcga tgaaaacgtt tcagtttgct
    10081 catggaaaac ggtgtaacaa gggtgaacac tatcccatat caccagctca ccgtctttca
    10141 ttgccatacg gaattccgga tgagcattca tcaggcgggc aagaatgtga ataaaggccg
    10201 gataaaactt gtgcttattt ttctttacgg tctttaaaaa ggccgtaata tccagctgaa
    10261 cggtctggtt ataggtacat tgagcaactg actgaaatgc ctcaaaatgt tctttacgat
    10321 gccattggga tatatcaacg gtggtatatc cagtgatttt tttctccatt ttagcttcct
    10381 tagctcctga aaatctcgat aactcaaaaa atacgcccgg tagtgatctt atttcattat
    10441 ggtgaaagtt ggaacctctt acgtgccgat caacgtctca ttttcgccag atatcgacgt
    10501 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
    10561 tcgtcttcac
//
