LOCUS       Exported                9465 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pTn7oluxG4, complete sequence.
ACCESSION   KF532962
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9465)
  AUTHORS   Kvtiko BH, Bruckbauer ST, Karkhoff-Schweizer RR, Schweizer HP.
  TITLE     Tn5/7 lux: A versatile vector for the location, capture, and 
            utilization of Gram negative promoters
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9465)
  AUTHORS   Kvtiko BH, Bruckbauer ST, Karkhoff-Schweizer RR, Schweizer HP.
  TITLE     Direct Submission
  JOURNAL   Submitted (09-AUG-2013) Microbiology Immunology and Pathology, 
            Colorado State University, IDRC at Foothills Campus, Campus Delivery
            0922, Fort Collins, CO 80523-0922, USA
REFERENCE   3  (bases 1 to 9465)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..9465
                     /organism="Cloning vector pTn7oluxG4"
                     /mol_type="other DNA"
                     /db_xref="taxon:1424481"
     misc_feature    219..480
                     /label=oriT
                     /note="oriT"
     oriT            365..473
                     /note="incP origin of transfer"
     rep_origin      504..885
                     /label=R6K oriV
                     /note="R6K oriV"
     rep_origin      524..880
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     misc_feature    1096..1261
                     /label=Tn7L
                     /note="Tn7L"
     mobile_element  1096..1261
                     /mobile_element_type="transposon:Tn7"
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     protein_bind    1292..1316
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    1341..1557
                     /label=PompA
                     /note="PompA"
     operon          1581..7378
                     /operon="luxCDABE operon"
                     /label=luxCDABE operon
     CDS             1581..3023
                     /codon_start=1
                     /operon="luxCDABE operon"
                     /gene="luxC"
                     /product="LuxC"
                     /label=luxC
                     /note="confers luminescent phenotype"
                     /protein_id="AHA61749.1"
                     /translation="MTKKISFIINGQVEIFPESDDLVQSINFGDNSVYLPILNDSHVKN
                     IIDCNGNNELRLHNIVNFLYTVGQRWKNEEYSRRRTYIRDLKKYMGYSEEMAKLEANWI
                     SMILCSKGGLYDVVENELGSRHIMDEWLPQDESYVRAFPKGKSVHLLAGNVPLSGIMSI
                     LRAILTKNQCIIKTSSTDPFTANALALSFIDVDPNHPITRSLSVIYWPHQGDTSLAKEI
                     MRHADVIVAWGGPDAINWAVEHAPSYADVIKFGSKKSLCIIDNPVDLTSAATGAAHDVC
                     FYDQRACFSAQNIYYMGNHYEEFKLALIEKLNLYAHILPNAKKDFDEKAAYSLVQKESL
                     FAGLKVEVDIHQRWMIIESNAGVEFNQPLGRCVYLHHVDNIEQILPYVQKNKTQTISIF
                     PWESSFKYRDALALKGAERIVEAGMNNIFRVGGSHDGMRPLQRLVTYISHERPSNYTAK
                     DVAVEIEQTRFLEEDKFLVFVP"
     gene            1581..3023
                     /operon="luxCDABE operon"
                     /gene="luxC"
                     /label=luxC
     CDS             3035..3958
                     /codon_start=1
                     /operon="luxCDABE operon"
                     /gene="luxD"
                     /product="LuxD"
                     /label=luxD
                     /note="confers luminescent phenotype"
                     /protein_id="AHA61750.1"
                     /translation="MENESKYKTIDHVICVEGNKKIHVWETLPEENSPKRKNAIIIASG
                     FARRMDHFAGLAEYLSRNGFHVIRYDSLHHVGLSSGTIDEFTMSIGKQSLLAVVDWLTT
                     RKINNFGMLASSLSARIAYASLSEINASFLITAVGVVNLRYSLERALGFDYLSLPINEL
                     PDNLDFEGHKLGAEVFARDCLDFGWEDLASTINNMMYLDIPFIAFTANNDNWVKQDEVI
                     TLLSNIRSNRCKIYSLLGSSHDLSENLVVLRNFYQSVTKAAIAMDNDHLDIDVDITEPS
                     FEHLTIATVNERRMRIEIENQAISLS"
     gene            3035..3958
                     /operon="luxCDABE operon"
                     /gene="luxD"
                     /label=luxD
     CDS             4007..5089
                     /codon_start=1
                     /operon="luxCDABE operon"
                     /gene="luxA"
                     /product="LuxA"
                     /label=luxA
                     /note="confers luminescent phenotype"
                     /protein_id="AHA61751.1"
                     /translation="MKFGNFLLTYQPPQFSQTEVMKRLVKLGRISEECGFDTVWLLEHH
                     FTEFGLLGNPYVAAAYLLGATKKLNVGTAAIVLPTAHPVRQLEDVNLLDQMSKGRFRFG
                     ICRGLYNKDFRVFGTDMNNSRALAECWYGLIKNGMTEGYMEADNEHIKFHKVKVNPAAY
                     SRGGAPVYVVAESASTTEWAAQFGLPMILSWIINTNEKKAQLELYNEVAQEYGHDIHNI
                     DHCLSYITSVDHDSIKAKEICRKFLGHWYDSYVNATTIFDDSDQTRGYDFNKGQWRDFV
                     LKGHKDTNRRIDYSYEINPVGTPQECIDIIQKDIDATGISNICCGFEANGTVDEIIASM
                     KLFQSDVMPFLKEKQRSLLY"
     gene            4007..5089
                     /operon="luxCDABE operon"
                     /gene="luxA"
                     /label=luxA
     CDS             5104..6087
                     /codon_start=1
                     /operon="luxCDABE operon"
                     /gene="luxB"
                     /product="LuxB"
                     /label=luxB
                     /note="confers luminescent phenotype"
                     /protein_id="AHA61752.1"
                     /translation="MKFGLFFLNFINSTTVQEQSIVRMQEITEYVDKLNFEQILVYENH
                     FSDNGVVGAPLTVSGFLLGLTEKIKIGSLNHIITTHHPVAIAEEACLLDQLSEGRFILG
                     FSDCEKKDEMHFFNRPVEYQQQLFEECYEIINDALTTGYCNPDNDFYSFPKISVNPHAY
                     TPGGPRKYVTATSHHIVEWAAKKGIPLIFKWDDSNDVRYEYAERYKAVADKYDVDLSEI
                     DHQLMILVNYNEDSNKAKQETRAFISDYVLEMHPNENFENKLEEIIAENAVGNYTECIT
                     AAKLAIEKCGAKSVLLSFEPMNDLMSQKNVINIVDDNIKKYHMEYT"
     gene            5104..6087
                     /operon="luxCDABE operon"
                     /gene="luxB"
                     /label=luxB
     CDS             6194..7378
                     /codon_start=1
                     /operon="luxCDABE operon"
                     /gene="luxE"
                     /product="LuxE"
                     /label=luxE
                     /note="confers luminescent phenotype"
                     /protein_id="AHA61753.1"
                     /translation="MKGIKEYDSSAAILSNIILRSKTGMTSYVDKQEITASSEIDDLIF
                     SSDPLVWSYDEQEKIRKKLVLDAFRNHYKHCREYRHYCQAHKVDDNITEIDDIPVFPTS
                     VFKFTRLLTSQENEIESWFTSSGTNGLKSQVARDRLSIERLLGSVSYGMKYVGSWFDHQ
                     IELVNLGPDRFNAHNIWFKYVMSLVELLYPTTFTVTEERIDFVKTLNSLERIKNQGKDL
                     CLIGSPYFIYLLCHYMKDKKISFSGDKSLYIITGGGWKSYEKESLKRDDFNHLLFDTFN
                     LSDISQIRDIFNQVELNTCFFEDEMQRKHVPPWVYARALDPETLKPVPDGTPGLMSYMD
                     ASATSYPAFIVTDDVGIISREYGKYPGVLVEILRRVNTRTQKGCALSLTEAFDS"
     gene            6194..7378
                     /operon="luxCDABE operon"
                     /gene="luxE"
                     /label=luxE
     CDS             6266..7378
                     /codon_start=1
                     /gene="luxE"
                     /product="LuxE"
                     /label=LuxE
                     /note="from the Photorhabdus luminescens bioluminescence 
                     operon"
                     /translation="MTSYVDKQEITASSEIDDLIFSSDPLVWSYDEQEKIRKKLVLDAF
                     RNHYKHCREYRHYCQAHKVDDNITEIDDIPVFPTSVFKFTRLLTSQENEIESWFTSSGT
                     NGLKSQVARDRLSIERLLGSVSYGMKYVGSWFDHQIELVNLGPDRFNAHNIWFKYVMSL
                     VELLYPTTFTVTEERIDFVKTLNSLERIKNQGKDLCLIGSPYFIYLLCHYMKDKKISFS
                     GDKSLYIITGGGWKSYEKESLKRDDFNHLLFDTFNLSDISQIRDIFNQVELNTCFFEDE
                     MQRKHVPPWVYARALDPETLKPVPDGTPGLMSYMDASATSYPAFIVTDDVGIISREYGK
                     YPGVLVEILRRVNTRTQKGCALSLTEAFDS"
     protein_bind    complement(7413..7437)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_recomb     7493..7540
                     /label=FRT
                     /note="FRT"
     protein_bind    complement(7493..7540)
                     /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        7581..7609
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             7798..8331
                     /codon_start=1
                     /gene="aaCI"
                     /product="AacCI"
                     /label=aaCI
                     /note="gentamicin resistance protein"
                     /protein_id="AHA61748.1"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     gene            7798..8331
                     /gene="aaCI"
                     /label=aaCI
     misc_recomb     8461..8508
                     /label=FRT
                     /note="FRT"
     protein_bind    complement(8461..8508)
                     /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)."
     regulatory      8549..8646
                     /regulatory_class="terminator"
                     /note="T1"
     terminator      8554..8640
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     regulatory      8743..8837
                     /regulatory_class="terminator"
                     /note="T0"
     terminator      8743..8837
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(8865..8881)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    8886..9084
                     /label=Tn7R
                     /note="Tn7R"