LOCUS       Exported               11266 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Fusion vector pFRT1-lux-Tel, complete sequence.
ACCESSION   FJ455410
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11266)
  AUTHORS   Kang Y, Norris MH, Barrett AR, Wilcox BA, Hoang TT.
  TITLE     Engineering of tellurite-resistant genetic tools for single-copy 
            chromosomal analysis of Burkholderia spp. and characterization of 
            the Burkholderia thailandensis betBA operon
  JOURNAL   Appl. Environ. Microbiol. 75 (12), 4015-4027 (2009)
  PUBMED    19376905
REFERENCE   2  (bases 1 to 11266)
  AUTHORS   Kang Y, Norris MH, Barrett AR, Videau PJA., Wilcox BA, Hoang TT.
  TITLE     Genetic Systems for Single Copy Analysis based on a Non-antibiotic 
            Selectable Marker in Burkholderia spp.
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 11266)
  AUTHORS   Kang Y, Norris MH, Barrett AR, Videau PJA., Wilcox BA, Hoang TT.
  TITLE     Direct Submission
  JOURNAL   Submitted (11-NOV-2008) Microbiology, University of Hawaii at Manoa,
            2538 McCarthy Mall, Snyder Hall 207, Honolulu, HI 96822, USA
REFERENCE   4  (bases 1 to 11266)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..11266
                     /organism="Fusion vector pFRT1-lux-Tel"
                     /mol_type="other DNA"
                     /note="FRT-lux fusion vector based on tellurite resistance"
                     /db_xref="taxon:579038"
     oriT            190..299
                     /note="incP origin of transfer"
     oriT            200..429
                     /label=origin of transfer for conjugation
                     /note="origin of transfer for conjugation"
     regulatory      720..1045
                     /regulatory_class="terminator"
                     /note="T1T2"
     terminator      720..747
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      839..925
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1546..2658)
                     /codon_start=1
                     /gene="luxE"
                     /product="LuxE"
                     /label=luxE
                     /note="luminescens luciferase complex"
                     /protein_id="ACJ70719.1"
                     /translation="MTSYVDKQEITASSEIDDLIFSSDPLVWSYDEQEKIRKKLVLDAF
                     RNHYKHCREYRHYCQAHKVDDNITEIDDIPVFPTSVFKFTRLLTSQENEIESWFTSSGT
                     NGLKSQVARDRLSIERLLGSVSYGMKYVGSWFDHQIELVNLGPDRFNAHNIWFKYVMSL
                     VELLYPTTFTVTEERIDFVKTLNSLERIKNQGKDLCLIGSPYFIYLLCHYMKDKKISFS
                     GDKSLYIITGGGWKSYEKESLKRDDFNHLLFDTFNLSDISQIRDIFNQVELNTCFFEDE
                     MQRKHVPPWVYARALDPETLKPVPDGTPGLMSYMDASATSYPAFIVTDDVGIISREYGK
                     YPGVLVEILRRVNTRTQKGCALSLTEAFDS"
     gene            complement(1546..2658)
                     /gene="luxE"
                     /label=luxE
     CDS             complement(2837..3820)
                     /codon_start=1
                     /gene="luxB"
                     /product="LuxB"
                     /label=luxB
                     /note="luminescens luciferase complex"
                     /protein_id="ACJ70720.1"
                     /translation="MKFGLFFLNFINSTTVQEQSIVRMQEITEYVDKLNFEQILVYENH
                     FSDNGVVGAPLTVSGFLLGLTEKIKIGSLNHIITTHHPVAIAEEACLLDQLSEGRFILG
                     FSDCEKKDEMHFFNRPVEYQQQLFEECYEIINDALTTGYCNPDNDFYSFPKISVNPHAY
                     TPGGPRKYVTATSHHIVEWAAKKGIPLIFKWDDSNDVRYEYAERYKAVADKYDVDLSEI
                     DHQLMILVNYNEDSNKAKQETRAFISDYVLEMHPNENFENKLEEIIAENAVGNYTECIT
                     AAKLAIEKCGAKSVLLSFEPMNDLMSQKNVINIVDDNIKKYHMEYT"
     gene            complement(2837..3820)
                     /gene="luxB"
                     /label=luxB
     CDS             complement(3835..4917)
                     /codon_start=1
                     /gene="luxA"
                     /product="LuxA"
                     /label=luxA
                     /note="luminescens luciferase complex"
                     /protein_id="ACJ70721.1"
                     /translation="MKFGNFLLTYQPPQFSQTEVMKRLVKLGRISEECGFDTVWLLEHH
                     FTEFGLLGNPYVAAAYLLGATKKLNVGTAAIVLPTAHPVRQLEDVNLLDQMSKGRFRFG
                     ICRGLYNKDFRVFGTDMNNSRALAECWYGLIKNGMTEGYMEADNEHIKFHKVKVNPAAY
                     SRGGAPVYVVAESASTTEWAAQFGLPMILSWIINTNEKKAQLELYNEVAQEYGHDIHNI
                     DHCLSYITSVDHDSIKAKEICRKFLGHWYDSYVNATTIFDDSDQTRGYDFNKGQWRDFV
                     LKGHKDTNRRIDYSYEINPVGTPQECIDIIQKDIDATGISNICCGFEANGTVDEIIASM
                     KLFQSDVMPFLKEKQRSLLY"
     gene            complement(3835..4917)
                     /gene="luxA"
                     /label=luxA
     CDS             complement(4966..5889)
                     /codon_start=1
                     /gene="luxD"
                     /product="LuxD"
                     /label=luxD
                     /note="luminescens luciferase complex"
                     /protein_id="ACJ70722.1"
                     /translation="MENESKYKTIDHVICVEGNKKIHVWETLPEENSPKRKNAIIIASG
                     FARRMDHFAGLAEYLSRNGFHVIRYDSLHHVGLSSGTIDEFTMSIGKQSLLAVVDWLTT
                     RKINNFGMLASSLSARIAYASLSEINASFLITAVGVVNLRYSLERALGFDYLSLPINEL
                     PDNLDFEGHKLGAEVFARDCLDFGWEDLASTINNMMYLDIPFIAFTANNDNWVKQDEVI
                     TLLSNIRSNRCKIYSLLGSSHDLSENLVVLRNFYQSVTKAAIAMDNDHLDIDVDITEPS
                     FEHLTIATVNERRMRIEIENQAISLS"
     gene            complement(4966..5889)
                     /gene="luxD"
                     /label=luxD
     CDS             complement(5901..7343)
                     /codon_start=1
                     /gene="luxC"
                     /product="LuxC"
                     /label=luxC
                     /note="luminescens luciferase complex"
                     /protein_id="ACJ70723.1"
                     /translation="MTKKISFIINGQVEIFPESDDLVQSINFGDNSVYLPILNDSHVKN
                     IIDCNGNNELRLHNIVNFLYTVGQRWKNEEYSRRRTYIRDLKKYMGYSEEMAKLEANWI
                     SMILCSKGGLYDVVENELGSRHIMDEWLPQDESYVRAFPKGKSVHLLAGNVPLSGIMSI
                     LRAILTKNQCIIKTSSTDPFTANALALSFIDVDPNHPITRSLSVIYWPHQGDTSLAKEI
                     MRHADVIVAWGGPDAINWAVEHAPSYADVIKFGSKKSLCIIDNPVDLTSAATGAAHDVC
                     FYDQRACFSAQNIYYMGNHYEEFKLALIEKLNLYAHILPNAKKDFDEKAAYSLVQKESL
                     FAGLKVEVDIHQRWMIIESNAGVEFNQPLGRCVYLHHVDNIEQILPYVQKNKTQTISIF
                     PWESSFKYRDALALKGAERIVEAGMNNIFRVGGSHDGMRPLQRLVTYISHERPSNYTAK
                     DVAVEIEQTRFLEEDKFLVFVP"
     gene            complement(5901..7343)
                     /gene="luxC"
                     /label=luxC
     misc_feature    complement(7391..7464)
                     /label=FRT
                     /note="FRT"
     protein_bind    7400..7447
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     promoter        7558..7586
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     regulatory      7710..7741
                     /regulatory_class="terminator"
                     /note="tonB"
     terminator      7710..7741
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(7759..8712)
                     /codon_start=1
                     /gene="telB"
                     /product="TelB"
                     /label=telB
                     /note="tellurite resistance"
                     /protein_id="ACJ70724.1"
                     /translation="MNATNTDVFAQVGGLEARGAKMKKRGTRFLIAALAVLAIAGIGAV
                     TGWAISPSATPGSIDVPQVLASTFSDQVPGSEGGGLGGGLPFTSAVGAFTDFMAGPAIF
                     TLGILGIVVAGAVLVFGGEFCGFVRSVCMMVIAVSMIFVSSNLVKGILGGDHDAGPAEP
                     SPRARFMAAVEAKDFARVQELIEARGAKSAADYVLAQLAVAEGLDRKPGARVVVGKAAG
                     SMAMPPAALGFTPRGEAAYAIERSAYGEPRSSIAKQYQQEWNRKAATWWAMAGVAGIIG
                     AILAAAATGFVGLAVSIRNRVKRVRDLLVMEPGAEP"
     gene            complement(7759..8712)
                     /gene="telB"
                     /label=telB
     CDS             complement(8709..9845)
                     /codon_start=1
                     /gene="telA"
                     /product="TelA"
                     /label=telA
                     /note="tellurite resistance"
                     /protein_id="ACJ70725.1"
                     /translation="MNALKTTHDAKAPIVAFDMTPATLRELGLQESDVPEVHAVAQRIE
                     VGSPQTVAEFGRDVAEHTSRYADSLLDQVRNSDLDEAGEKLTQVVAKARSLNVGPLSDN
                     RSRLPLIGPLIDRFRVRSTGFMARFDTTREQIEHLVSEVQTTQQGIAQRNASLDEMFAA
                     VREEHRLLGVHIAAGKVRLAELREQAEGLRGNVGNDPGRVQELADLDAMVANLDKRIGD
                     LIALQHSAMQSLPTIRMIQANNQMLVDKFHTIREITVPAWKRQFMLALSLNEQKNAVEL
                     ATAIDDTTNDLMKRNAALLHRTSVETAKENQRLVIDVDTLKQVQTTLIKTVEDVIRIQQ
                     EGVQKRKDAEKQIAAMRGDLQAKLTRQPVRELAQQESV"
     gene            complement(8709..9845)
                     /gene="telA"
                     /label=telA
     CDS             complement(9863..10636)
                     /codon_start=1
                     /gene="kilA"
                     /product="KilA"
                     /label=kilA
                     /note="tellurite resistance"
                     /protein_id="ACJ70726.1"
                     /translation="MEEQSVNMARLKGEVLPALFASPATIGEYGAGIDGADSLNELSNL
                     MEHGAVAALADKISQIVAKLADADPRKIAEKPTWFEKMLGREVERQVRYQVARKTLDQL
                     LDEAEGVAQRVRDTLRALDDMLNTHEAEVDRLRAYIQAGREFLDENPEAGAAKAGVIEF
                     DKPRERFARKLANLATLMASHEMSVTQMKLTRAQAVDMLDRFSETASVLVPVWRQHTLA
                     LITTKNMNPAMVAEAAKAHQALMRSLSQSLEGINQ"
     gene            complement(9863..10636)
                     /gene="kilA"
                     /label=kilA
     RBS             10643..10665
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     regulatory      complement(10676..10717)
                     /regulatory_class="promoter"
                     /note="derived from Burkholderia cenocepacia rpsL promoter"
     rep_origin      10858..11216
                     /label=R6K origin of replication
                     /note="R6K origin of replication"