LOCUS       Exported               10570 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Regulator-reporter vector pWUR770, complete sequence.
ACCESSION   KX670547
VERSION     .
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)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..10570
                     /organism="Regulator-reporter vector pWUR770"
                     /mol_type="other DNA"
                     /db_xref="taxon:1979215"
     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="ARH53398.1"
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     gene            107..985
                     /gene="araC"
                     /label=araC
     CDS             complement(992..2083)
                     /codon_start=1
                     /gene="leuB"
                     /product="3-isopropylmalate dehydrogenase"
                     /label=leuB
                     /note="LeuB; leucine auxotrophy complementation"
                     /protein_id="ARH53399.1"
                     /translation="MSKNYHIAVLPGDGIGPEVMTQALKVLDAVRNRFAMRITTSHYDV
                     GGAAIDNHGQPLPPATVEGCEQADAVLFGSVGGPKWEHLPPDQQPERGALLPLRKHFKL
                     FSNLRPAKLYQGLEAFCPLRADIAANGFDILCVRELTGGIYFGQPKGREGSGQYEKAFD
                     TEVYHRFEIERIARIAFESARKRRHKVTSIDKANVLQSSILWREIVNEIATEYPDVELA
                     HMYIDNATMQLIKDPSQFDVLLCSNLFGDILSDECAMITGSMGMLPSASLNEQGFGLYE
                     PAGGSAPDIAGKNIANPIAQILSLALLLRYSLDADDAACAIERAINRALEEGIRTGDLA
                     RGAAAVSTDEMGDIIARYVAEGV"
     gene            complement(992..2083)
                     /gene="leuB"
                     /label=leuB
     regulatory      complement(2090..2095)
                     /regulatory_class="ribosome_binding_site"
                     /note="RBS"
     regulatory      complement(2104..2414)
                     /regulatory_class="promoter"
                     /note="PBAD-adapt"
     rep_origin      complement(2421..3215)
                     /direction=LEFT
                     /label=p15A
                     /note="p15A"
     rep_origin      2521..3066
                     /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      3336..3646
                     /regulatory_class="promoter"
                     /note="PBAD-adapt"
     regulatory      3653..3658
                     /regulatory_class="ribosome_binding_site"
                     /note="RBS"
     CDS             3665..5107
                     /codon_start=1
                     /gene="luxC"
                     /product="fatty acid reductase"
                     /label=luxC
                     /note="LuxC; formation of aldehyde substrate for 
                     luciferase"
                     /protein_id="ARH53400.1"
                     /translation="MTKKISFIINGQVEIFPEGDDLVQSINFGDNSVYLPILNDSHVKN
                     IIDCNGNNELRLHNIVNFLYTVGQRWKNEEYSRRRTYIRDLKKYMGYSEEMAKLEANWI
                     SMILCSKGGLYDVVENELGSRHIMDEWLPQDESYVRAFPKGKSVHLLAGNVPLSGIMSI
                     LRAILTKNQCIIKTSSTDPFTANALALSFIDVDPNHPITRSLSVIYWPHQGDTSLAKEI
                     MQHADVIVAWGGPDAINWAVEHAPSYADVIKFGSKKSLCIIDNPVDLTSAATGAAHDVC
                     FYDQRACFSAQNIYYMGNHYEEFKLALIEKLNLYAHILPNAKKDFDEKAAYSLVQKESL
                     FAGLKVEVDIHQRWMIIESNAGVEFNQPLGRCVYLHHVDNIEQILPYVQKNKTQTISIF
                     PWESSFKYRDALALKGAERIVEAGMNNIFRVGGSHDGMRPLQRLVTYISHERPSNYTAK
                     DVAVEIEQTRFLEEDKFLVFVP"
     gene            3665..5107
                     /gene="luxC"
                     /label=luxC
     CDS             5120..6043
                     /codon_start=1
                     /gene="luxD"
                     /product="acyl transferase"
                     /label=luxD
                     /note="LuxD; formation of aldehyde substrate for 
                     luciferase"
                     /protein_id="ARH53401.1"
                     /translation="MENESKYKTIDHVICVEGNKKIHVWETLPEENSPKRKNAIIIASG
                     FARRMDHFAGLAEYLSRNGFHVIRYDSLHHVGLSSGTIDEFTMSIGKQSLLAVVDWLTT
                     RKINNFGMLASSLSARIAYASLSEINASFLITAVGVVNLRYSLERALGFDYLSLPINEL
                     PNNLDFEGHKLGAEVFARDCLDFGWEDLASTINNMMYLDIPFIAFTANNDNWVKQDEVI
                     TLLSNIRSNRCKIYSLLGSSHDLSENLVVLRNFYQSVTKAAIAMDNDHLDIDVDITEPS
                     FEHLTIATVNERRMRIEIENQAISLS"
     gene            5120..6043
                     /gene="luxD"
                     /label=luxD
     CDS             6092..7174
                     /codon_start=1
                     /gene="luxA"
                     /product="luciferase alpha subunit"
                     /label=luxA
                     /note="LuxA"
                     /protein_id="ARH53402.1"
                     /translation="MKFGNFLLTYQPPQFSQTEVMKRLVKLGRISEECGFDTVWLLEHH
                     FTEFGLLGNPYVAAAYLLGATKKLNVGTAAIVLPTAHPVRQLEDVNLLDQMSKGRFRFG
                     ICRGLYNKDFRVFGTDMNNSRALAECWYGLIKNGMTEGYMEADNEHIKFHKVKVNPAAY
                     SRGGAPVYVVAESASTTEWAAQFGLPMILSWIINTNEKKAQLELYNEVAQEYGHDIHNI
                     DHCLSYITSVDHDSIKAKEICRKFLGHWYDSYVNATTIFDDSDQTRGYDFNKGQWRDFV
                     LKGHKDTNRRIDYSYEINPVGTPQECIDIIQKDIDATGISNICCGFEANGTVDEIIASM
                     KLFQSDVMPFLKEKQRSLLY"
     gene            6092..7174
                     /gene="luxA"
                     /label=luxA
     CDS             7189..8172
                     /codon_start=1
                     /gene="luxB"
                     /product="luciferase beta subunit"
                     /label=luxB
                     /note="LuxB"
                     /protein_id="ARH53403.1"
                     /translation="MKFGLFFLNFINSTTVQEQSIVRMQEITEYVDKLNFEQILVYENH
                     FSDNGVVGAPLTVSGFLLGLTEKIKIGSLNHIITTHHPVRIAEEACLLDQLSEGRFILG
                     FSDCEKKDEMHFFNRPVEYQQQLFEECYEIINDALTTGYCNPDNDFYSFPKISVNPHAY
                     TPGGPRKYVTATSHHIVEWAAKKGIPLIFKWDDSNDVRYEYAERYKAVADKYDVDLSEI
                     DHQLMILVNYNEDSNKAKQETRAFISDYVLEMHPNENFENKLEEIIAENAVGNYTECIT
                     AAKLAIEKCGAKSVLLSFEPMNDLMSQKNVINIVDDNIKKYHMEYT"
     gene            7189..8172
                     /gene="luxB"
                     /label=luxB
     CDS             8351..9463
                     /codon_start=1
                     /gene="luxE"
                     /product="acyl-protein synthetase"
                     /label=luxE
                     /note="LuxE; formation of aldehyde substrate for 
                     luciferase"
                     /protein_id="ARH53404.1"
                     /translation="MTSYVDKQEITASSEIDDLIFSSDPLVWSYDEQEKIRKKLVLDAF
                     RNHYKHCREYRHYCQAHKVDDNITEIDDIPVFPTSVFKFTRLLTSQENEIESWFTSSGT
                     NGLKSQVARDRLSIERLLGSVSYGMKYVGSWFDHQIELVNLGPDRFNAHNIWFKYVMSL
                     VELLYPTTFTVTEERIDFVKTLNSLERIKNQGKDLCLIGSPYFIYLLCHYMKDKKISFS
                     GDKSLYIITGGGWKSYEKESLKRDDFNHLLFDTFNLSDISQIRDIFNQVELNTCFFEDE
                     MQRKHVPPWVYARALDPETLKPVPDGTPGLMSYMDASATSYPAFIVTDDVGIISREYGK
                     YPGVLVEILRRVNTRTQKGCALSLTEAFDS"
     gene            8351..9463
                     /gene="luxE"
                     /label=luxE
     regulatory      9478..9582
                     /regulatory_class="terminator"
                     /note="T1 terminator"
     terminator      9483..9569
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     regulatory      complement(9595..9689)
                     /regulatory_class="terminator"
                     /note="t0 terminator"
     terminator      9595..9689
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(9710..10369)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=cat
                     /note="CmR; chloramphenicol resistance protein"
                     /protein_id="ARH53405.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     gene            complement(9710..10369)
                     /gene="cat"
                     /label=cat
     promoter        complement(10370..10472)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"