LOCUS       Exported               10294 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Regulator-reporter vector pWUR772, complete sequence.
ACCESSION   KX670548
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10294)
  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 10294)
  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 10294)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..10294
                     /organism="Regulator-reporter vector pWUR772"
                     /mol_type="other DNA"
                     /db_xref="taxon:1979216"
     regulatory      10..87
                     /regulatory_class="promoter"
                     /note="PlacIq"
     promoter        10..87
                     /gene="lacI (mutant)"
                     /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
                     /regulatory_class="ribosome_binding_site"
                     /note="RBS"
     CDS             107..985
                     /codon_start=1
                     /gene="araC"
                     /product="AraC"
                     /label=araC
                     /note="transcriptional activator for the ara regulon"
                     /protein_id="ARH53406.1"
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     gene            107..985
                     /gene="araC"
                     /label=araC
     CDS             complement(992..1807)
                     /codon_start=1
                     /gene="kan"
                     /product="aminoglycoside 3-phosphotransferase"
                     /label=kan
                     /note="KmR; kanamycin resistance protein"
                     /protein_id="ARH53407.1"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     gene            complement(992..1807)
                     /gene="kan"
                     /label=kan
     regulatory      complement(1814..1819)
                     /regulatory_class="ribosome_binding_site"
                     /note="RBS"
     regulatory      complement(1828..2138)
                     /regulatory_class="promoter"
                     /note="PBAD-adapt"
     rep_origin      complement(2145..2939)
                     /direction=LEFT
                     /label=p15A
                     /note="p15A"
     rep_origin      2245..2790
                     /direction=RIGHT
                     /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      3060..3370
                     /regulatory_class="promoter"
                     /note="PBAD-adapt"
     regulatory      3377..3382
                     /regulatory_class="ribosome_binding_site"
                     /note="RBS"
     CDS             3389..4831
                     /codon_start=1
                     /gene="luxC"
                     /product="fatty acid reductase"
                     /label=luxC
                     /note="LuxC; formation of aldehyde substrate for 
                     luciferase"
                     /protein_id="ARH53408.1"
                     /translation="MTKKISFIINGQVEIFPEGDDLVQSINFGDNSVYLPILNDSHVKN
                     IIDCNGNNELRLHNIVNFLYTVGQRWKNEEYSRRRTYIRDLKKYMGYSEEMAKLEANWI
                     SMILCSKGGLYDVVENELGSRHIMDEWLPQDESYVRAFPKGKSVHLLAGNVPLSGIMSI
                     LRAILTKNQCIIKTSSTDPFTANALALSFIDVDPNHPITRSLSVIYWPHQGDTSLAKEI
                     MQHADVIVAWGGPDAINWAVEHAPSYADVIKFGSKKSLCIIDNPVDLTSAATGAAHDVC
                     FYDQRACFSAQNIYYMGNHYEEFKLALIEKLNLYAHILPNAKKDFDEKAAYSLVQKESL
                     FAGLKVEVDIHQRWMIIESNAGVEFNQPLGRCVYLHHVDNIEQILPYVQKNKTQTISIF
                     PWESSFKYRDALALKGAERIVEAGMNNIFRVGGSHDGMRPLQRLVTYISHERPSNYTAK
                     DVAVEIEQTRFLEEDKFLVFVP"
     gene            3389..4831
                     /gene="luxC"
                     /label=luxC
     CDS             4844..5767
                     /codon_start=1
                     /gene="luxD"
                     /product="acyl transferase"
                     /label=luxD
                     /note="LuxD; formation of aldehyde substrate for 
                     luciferase"
                     /protein_id="ARH53409.1"
                     /translation="MENESKYKTIDHVICVEGNKKIHVWETLPEENSPKRKNAIIIASG
                     FARRMDHFAGLAEYLSRNGFHVIRYDSLHHVGLSSGTIDEFTMSIGKQSLLAVVDWLTT
                     RKINNFGMLASSLSARIAYASLSEINASFLITAVGVVNLRYSLERALGFDYLSLPINEL
                     PNNLDFEGHKLGAEVFARDCLDFGWEDLASTINNMMYLDIPFIAFTANNDNWVKQDEVI
                     TLLSNIRSNRCKIYSLLGSSHDLSENLVVLRNFYQSVTKAAIAMDNDHLDIDVDITEPS
                     FEHLTIATVNERRMRIEIENQAISLS"
     gene            4844..5767
                     /gene="luxD"
                     /label=luxD
     CDS             5816..6898
                     /codon_start=1
                     /gene="luxA"
                     /product="luciferase alpha subunit"
                     /label=luxA
                     /note="LuxA"
                     /protein_id="ARH53410.1"
                     /translation="MKFGNFLLTYQPPQFSQTEVMKRLVKLGRISEECGFDTVWLLEHH
                     FTEFGLLGNPYVAAAYLLGATKKLNVGTAAIVLPTAHPVRQLEDVNLLDQMSKGRFRFG
                     ICRGLYNKDFRVFGTDMNNSRALAECWYGLIKNGMTEGYMEADNEHIKFHKVKVNPAAY
                     SRGGAPVYVVAESASTTEWAAQFGLPMILSWIINTNEKKAQLELYNEVAQEYGHDIHNI
                     DHCLSYITSVDHDSIKAKEICRKFLGHWYDSYVNATTIFDDSDQTRGYDFNKGQWRDFV
                     LKGHKDTNRRIDYSYEINPVGTPQECIDIIQKDIDATGISNICCGFEANGTVDEIIASM
                     KLFQSDVMPFLKEKQRSLLY"
     gene            5816..6898
                     /gene="luxA"
                     /label=luxA
     CDS             6913..7896
                     /codon_start=1
                     /gene="luxB"
                     /product="luciferase beta subunit"
                     /label=luxB
                     /note="LuxB"
                     /protein_id="ARH53411.1"
                     /translation="MKFGLFFLNFINSTTVQEQSIVRMQEITEYVDKLNFEQILVYENH
                     FSDNGVVGAPLTVSGFLLGLTEKIKIGSLNHIITTHHPVRIAEEACLLDQLSEGRFILG
                     FSDCEKKDEMHFFNRPVEYQQQLFEECYEIINDALTTGYCNPDNDFYSFPKISVNPHAY
                     TPGGPRKYVTATSHHIVEWAAKKGIPLIFKWDDSNDVRYEYAERYKAVADKYDVDLSEI
                     DHQLMILVNYNEDSNKAKQETRAFISDYVLEMHPNENFENKLEEIIAENAVGNYTECIT
                     AAKLAIEKCGAKSVLLSFEPMNDLMSQKNVINIVDDNIKKYHMEYT"
     gene            6913..7896
                     /gene="luxB"
                     /label=luxB
     CDS             8075..9187
                     /codon_start=1
                     /gene="luxE"
                     /product="acyl-protein synthetase"
                     /label=luxE
                     /note="LuxE; formation of aldehyde substrate for 
                     luciferase"
                     /protein_id="ARH53412.1"
                     /translation="MTSYVDKQEITASSEIDDLIFSSDPLVWSYDEQEKIRKKLVLDAF
                     RNHYKHCREYRHYCQAHKVDDNITEIDDIPVFPTSVFKFTRLLTSQENEIESWFTSSGT
                     NGLKSQVARDRLSIERLLGSVSYGMKYVGSWFDHQIELVNLGPDRFNAHNIWFKYVMSL
                     VELLYPTTFTVTEERIDFVKTLNSLERIKNQGKDLCLIGSPYFIYLLCHYMKDKKISFS
                     GDKSLYIITGGGWKSYEKESLKRDDFNHLLFDTFNLSDISQIRDIFNQVELNTCFFEDE
                     MQRKHVPPWVYARALDPETLKPVPDGTPGLMSYMDASATSYPAFIVTDDVGIISREYGK
                     YPGVLVEILRRVNTRTQKGCALSLTEAFDS"
     gene            8075..9187
                     /gene="luxE"
                     /label=luxE
     regulatory      9202..9306
                     /regulatory_class="terminator"
                     /note="T1 terminator"
     terminator      9207..9293
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     regulatory      complement(9319..9413)
                     /regulatory_class="terminator"
                     /note="t0 terminator"
     terminator      9319..9413
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(9434..10093)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=cat
                     /note="CmR; chloramphenicol resistance protein"
                     /protein_id="ARH53413.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     gene            complement(9434..10093)
                     /gene="cat"
                     /label=cat
     promoter        complement(10094..10196)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"