Basic Vector Information
pTn5.7luxK4 vector Vector Map
Plasmid Resuspension Protocol:
1. Centrifuge at 5,000×g for 5 min.
2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.
3. Close the tube and incubate for 10 minutes at room temperature.
4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.
5.Store the plasmid at -20 ℃.
pTn5.7luxK4 vector Sequence
LOCUS 40924_43638 11324 bp DNA circular SYN 18-DEC-2018 DEFINITION Cloning vector pTn5.7luxK4, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 11324) 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 TITLE Tn5/7 lux: A versatile vector for the location, capture, and utilization of Gram negative promoters JOURNAL Unpublished REFERENCE 2 (bases 1 to 11324) AUTHORS Kvtiko BH, Bruckbauer ST, Karkhoff-Schweizer RR, Schweizer HP.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 11324) TITLE Direct Submission REFERENCE 4 (bases 1 to 11324) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Unpublished" COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted (09-AUG-2013) Microbiology Immunology and Pathology, Colorado State University, IDRC at Foothills Campus, Campus Delivery 0922, Fort Collins, CO 80523-0922, USA" COMMENT SGRef: number: 3; type: "Journal Article" COMMENT ##Assembly-Data-START## Sequencing Technology :: Sanger dideoxy sequencing ##Assembly-Data-END## FEATURES Location/Qualifiers source 1..11324 /mol_type="other DNA" /organism="synthetic DNA construct" oriT 365..473 /label=oriT /note="incP origin of transfer" rep_origin 496..880 /label=R6K gamma ori /note="gamma replication origin from E. coli plasmid R6K; requires the R6K initiator protein pi for replication" mobile_element 1096..1261 /label=Tn7L /note="mini-Tn7 element (left end of the Tn7 transposon)" misc_feature complement(1276..1294) /label=Tn5 ME /note="hyperactive mosaic end for Tn5 transposase recognition (Reznikoff et al., 2004)" promoter complement(1295..1399) /gene="bla" /label=AmpR promoter CDS complement(1449..2876) /codon_start=1 /label=Tn5 transposase /note="transposase from the bacterial Tn5 transposon (Reznikoff, 1993)" /translation="MITSALHRAADWAKSVFSSAALGDPRRTARLVNVAAQLAKYSGKS IPISSEGSEAAQEGAYRFIRNPNVSAEAIRKAGAMQTVKLAQEFPELLAIEDTTSLSYR HQVAEELGKLGSIQDKSRGWWVHSVLLLEATTFRTVGLLHQEWWMRPDDPADADEKESG KWLAAAATSRLRMGSMMSNVIAVCDREADIHAYLQDKLAHNERFVVRSKHPRKDVESGL YLYDHLKNQPELGGYQISIPQKGVVDKRGKRKNRPARKASLSLRSGRITLKQGNITLNA VLAEEINPPKGETPLKWLLLTSEPVESLAQALRVIDIYTHRWRIEEFHKAWKTGAGAER QRMEEPDNLERMVSILSFVAVRLLQLRESFTLPQALRAQGLLKEAEHVESQSAETVLTP DECQLLGYLDKGKRKRKEKAGSLQWAYMAIARLGGFMDSKRTGIASWGALWEGWEALQS KLDGFLAAKDLMAQGIKI" misc_feature 3031..3049 /label=Tn5 ME /note="hyperactive mosaic end for Tn5 transposase recognition (Reznikoff et al., 2004)" CDS 3082..4521 /codon_start=1 /label=LuxC /note="LuxC fatty acid reductase" /translation="MTKKISFIINGQVEIFPESDDLVQSINFGDNSVYLPILNDSHVKN IIDCNGNNELRLHNIVNFLYTVGQRWKNEEYSRRRTYIRDLKKYMGYSEEMAKLEANWI SMILCSKGGLYDVVENELGSRHIMDEWLPQDESYVRAFPKGKSVHLLAGNVPLSGIMSI LRAILTKNQCIIKTSSTDPFTANALALSFIDVDPNHPITRSLSVIYWPHQGDTSLAKEI MRHADVIVAWGGPDAINWAVEHAPSYADVIKFGSKKSLCIIDNPVDLTSAATGAAHDVC FYDQRACFSAQNIYYMGNHYEEFKLALIEKLNLYAHILPNAKKDFDEKAAYSLVQKESL FAGLKVEVDIHQRWMIIESNAGVEFNQPLGRCVYLHHVDNIEQILPYVQKNKTQTISIF PWESSFKYRDALALKGAERIVEAGMNNIFRVGGSHDGMRPLQRLVTYISHERPSNYTAK DVAVEIEQTRFLEEDKFLVFVP" CDS 4536..5456 /codon_start=1 /label=LuxD /note="LuxD acyltransferase" /translation="MENESKYKTIDHVICVEGNKKIHVWETLPEENSPKRKNAIIIASG FARRMDHFAGLAEYLSRNGFHVIRYDSLHHVGLSSGTIDEFTMSIGKQSLLAVVDWLTT RKINNFGMLASSLSARIAYASLSEINASFLITAVGVVNLRYSLERALGFDYLSLPINEL PDNLDFEGHKLGAEVFARDCLDFGWEDLASTINNMMYLDIPFIAFTANNDNWVKQDEVI TLLSNIRSNRCKIYSLLGSSHDLSENLVVLRNFYQSVTKAAIAMDNDHLDIDVDITEPS FEHLTIATVNERRMRIEIENQAISLS" CDS 5508..6587 /codon_start=1 /label=LuxA /note="LuxA luciferase subunit" /translation="MKFGNFLLTYQPPQFSQTEVMKRLVKLGRISEECGFDTVWLLEHH FTEFGLLGNPYVAAAYLLGATKKLNVGTAAIVLPTAHPVRQLEDVNLLDQMSKGRFRFG ICRGLYNKDFRVFGTDMNNSRALAECWYGLIKNGMTEGYMEADNEHIKFHKVKVNPAAY SRGGAPVYVVAESASTTEWAAQFGLPMILSWIINTNEKKAQLELYNEVAQEYGHDIHNI DHCLSYITSVDHDSIKAKEICRKFLGHWYDSYVNATTIFDDSDQTRGYDFNKGQWRDFV LKGHKDTNRRIDYSYEINPVGTPQECIDIIQKDIDATGISNICCGFEANGTVDEIIASM KLFQSDVMPFLKEKQRSLLY" CDS 6605..7585 /codon_start=1 /label=LuxB /note="LuxB luciferase subunit" /translation="MKFGLFFLNFINSTTVQEQSIVRMQEITEYVDKLNFEQILVYENH FSDNGVVGAPLTVSGFLLGLTEKIKIGSLNHIITTHHPVAIAEEACLLDQLSEGRFILG FSDCEKKDEMHFFNRPVEYQQQLFEECYEIINDALTTGYCNPDNDFYSFPKISVNPHAY TPGGPRKYVTATSHHIVEWAAKKGIPLIFKWDDSNDVRYEYAERYKAVADKYDVDLSEI DHQLMILVNYNEDSNKAKQETRAFISDYVLEMHPNENFENKLEEIIAENAVGNYTECIT AAKLAIEKCGAKSVLLSFEPMNDLMSQKNVINIVDDNIKKYHMEYT" CDS 7767..8876 /codon_start=1 /label=LuxE /note="LuxE" /translation="MTSYVDKQEITASSEIDDLIFSSDPLVWSYDEQEKIRKKLVLDAF RNHYKHCREYRHYCQAHKVDDNITEIDDIPVFPTSVFKFTRLLTSQENEIESWFTSSGT NGLKSQVARDRLSIERLLGSVSYGMKYVGSWFDHQIELVNLGPDRFNAHNIWFKYVMSL VELLYPTTFTVTEERIDFVKTLNSLERIKNQGKDLCLIGSPYFIYLLCHYMKDKKISFS GDKSLYIITGGGWKSYEKESLKRDDFNHLLFDTFNLSDISQIRDIFNQVELNTCFFEDE MQRKHVPPWVYARALDPETLKPVPDGTPGLMSYMDASATSYPAFIVTDDVGIISREYGK YPGVLVEILRRVNTRTQKGCALSLTEAFDS" protein_bind complement(8914..8938) /label=attB2 /note="recombination site for the Gateway(R) BP reaction" misc_recomb 8994..9041 /label=FRT /note="FRT" protein_bind complement(8994..9041) /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)." CDS 9427..10218 /codon_start=1 /label=NeoR/KanR /note="aminoglycoside phosphotransferase" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" protein_bind complement(10320..10367) /label=FRT /note="FLP-mediated recombination occurs in the 8-bp core sequence TCTAGAAA (Turan and Bode, 2011)." terminator complement(10413..10499) /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator complement(10602..10696) /label=lambda t0 terminator /note="transcription terminator from phage lambda" primer_bind complement(10724..10740) /label=KS primer /note="common sequencing primer, one of multiple similar variants" misc_feature 10745..10943 /label=Tn7R /note="Tn7R"
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