Basic Vector Information
| Basic Vector Information | |||
|---|---|---|---|
| Vector Name | pTn5.7luxK3 | Length | 11265 bp |
| Type | Promoter capture vector | Source | Bruckbauer ST, Kvitko BH, Karkhoff-Schweizer RR, Schweizer HP. |
pTn5.7luxK3 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 ℃.
pTn5.7luxK3 vector Sequence
LOCUS Exported 11265 bp ds-DNA circular SYN 18-DEC-2018
DEFINITION Promoter capture vector pTn5.7luxK3, complete sequence.
ACCESSION KC332286
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 11265)
AUTHORS Bruckbauer ST, Kvitko BH, 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JOURNAL Unpublished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 11265)
AUTHORS Bruckbauer ST, Kvitko BH, Karkhoff-Schweizer RR, Schweizer HP.
TITLE Direct Submission
JOURNAL Submitted (10-DEC-2012) 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 11265)
AUTHORS .
TITLE Direct Submission
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..11265
/organism="Promoter capture vector pTn5.7luxK3"
/mol_type="other DNA"
/db_xref="taxon:1303869"
oriT 219..480
oriT 365..473
/note="incP origin of transfer"
rep_origin 504..885
/map="R6K gamma"
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)"
misc_feature complement(1276..1294)
/label=Tn5 mosaic end
/note="Tn5 mosaic end"
promoter complement(1295..1399)
/gene="bla"
/label=AmpR promoter
CDS complement(1446..2888)
/codon_start=1
/gene="tnpA"
/product="TnpA"
/label=tnpA
/note="Tn5 transposase"
/protein_id="AGI04966.1"
/translation="MVTFMITSALHRAADWAKSVFSSAALGDPRRTARLVNVAAQLAKY
SGKSIPISSEGSEAAQEGAYRFIRNPNVSAEAIRKAGAMQTVKLAQEFPELLAIEDTTS
LSYRHQVAEELGKLGSIQDKSRGWWVHSVLLLEATTFRTVGLLHQEWWMRPDDPADADE
KESGKWLAAAATSRLRMGSMMSNVIAVCDREADIHAYLQDKLAHNERFVVRSKHPRKDV
ESGLYLYDHLKNQPELGGYQISIPQKGVVDKRGKRKNRPARKASLSLRSGRITLKQGNI
TLNAVLAEEINPPKGETPLKWLLLTSEPVESLAQALRVIDIYTHRWRIEEFHKAWKTGA
GAERQRMEEPDNLERMVSILSFVAVRLLQLRESFTLPQALRAQGLLKEAEHVESQSAET
VLTPDECQLLGYLDKGKRKRKEKAGSLQWAYMAIARLGGFMDSKRTGIASWGALWEGWE
ALQSKLDGFLAAKDLMAQGIKI"
gene complement(1446..2888)
/gene="tnpA"
/label=tnpA
misc_feature complement(3031..3049)
/label=Tn5 mosaic end
/note="Tn5 mosaic end"
CDS 3082..4524
/codon_start=1
/gene="luxC"
/product="LuxC"
/label=luxC
/note="confers luminescent phenotype"
/protein_id="AGI04967.1"
/translation="MTKKISFIINGQVEIFPESDDLVQSINFGDNSVYLPILNDSHVKN
IIDCNGNNELRLHNIVNFLYTVGQRWKNEEYSRRRTYIRDLKKYMGYSEEMAKLEANWI
SMILCSKGGLYDVVENELGSRHIMDEWLPQDESYVRAFPKGKSVHLLAGNVPLSGIMSI
LRAILTKNQCIIKTSSTDPFTANALALSFIDVDPNHPITRSLSVIYWPHQGDTSLAKEI
MRHADVIVAWGGPDAINWAVEHAPSYADVIKFGSKKSLCIIDNPVDLTSAATGAAHDVC
FYDQRACFSAQNIYYMGNHYEEFKLALIEKLNLYAHILPNAKKDFDEKAAYSLVQKESL
FAGLKVEVDIHQRWMIIESNAGVEFNQPLGRCVYLHHVDNIEQILPYVQKNKTQTISIF
PWESSFKYRDALALKGAERIVEAGMNNIFRVGGSHDGMRPLQRLVTYISHERPSNYTAK
DVAVEIEQTRFLEEDKFLVFVP"
gene 3082..4524
/gene="luxC"
/label=luxC
CDS 4536..5459
/codon_start=1
/gene="luxD"
/product="LuxD"
/label=luxD
/note="confers luminescent phenotype"
/protein_id="AGI04968.1"
/translation="MENESKYKTIDHVICVEGNKKIHVWETLPEENSPKRKNAIIIASG
FARRMDHFAGLAEYLSRNGFHVIRYDSLHHVGLSSGTIDEFTMSIGKQSLLAVVDWLTT
RKINNFGMLASSLSARIAYASLSEINASFLITAVGVVNLRYSLERALGFDYLSLPINEL
PDNLDFEGHKLGAEVFARDCLDFGWEDLASTINNMMYLDIPFIAFTANNDNWVKQDEVI
TLLSNIRSNRCKIYSLLGSSHDLSENLVVLRNFYQSVTKAAIAMDNDHLDIDVDITEPS
FEHLTIATVNERRMRIEIENQAISLS"
gene 4536..5459
/gene="luxD"
/label=luxD
CDS 5508..6590
/codon_start=1
/gene="luxA"
/product="LuxA"
/label=luxA
/note="confers luminescent phenotype"
/protein_id="AGI04969.1"
/translation="MKFGNFLLTYQPPQFSQTEVMKRLVKLGRISEECGFDTVWLLEHH
FTEFGLLGNPYVAAAYLLGATKKLNVGTAAIVLPTAHPVRQLEDVNLLDQMSKGRFRFG
ICRGLYNKDFRVFGTDMNNSRALAECWYGLIKNGMTEGYMEADNEHIKFHKVKVNPAAY
SRGGAPVYVVAESASTTEWAAQFGLPMILSWIINTNEKKAQLELYNEVAQEYGHDIHNI
DHCLSYITSVDHDSIKAKEICRKFLGHWYDSYVNATTIFDDSDQTRGYDFNKGQWRDFV
LKGHKDTNRRIDYSYEINPVGTPQECIDIIQKDIDATGISNICCGFEANGTVDEIIASM
KLFQSDVMPFLKEKQRSLLY"
gene 5508..6590
/gene="luxA"
/label=luxA
CDS 6605..7588
/codon_start=1
/gene="luxB"
/product="LuxB"
/label=luxB
/note="confers luminescent phenotype"
/protein_id="AGI04970.1"
/translation="MKFGLFFLNFINSTTVQEQSIVRMQEITEYVDKLNFEQILVYENH
FSDNGVVGAPLTVSGFLLGLTEKIKIGSLNHIITTHHPVAIAEEACLLDQLSEGRFILG
FSDCEKKDEMHFFNRPVEYQQQLFEECYEIINDALTTGYCNPDNDFYSFPKISVNPHAY
TPGGPRKYVTATSHHIVEWAAKKGIPLIFKWDDSNDVRYEYAERYKAVADKYDVDLSEI
DHQLMILVNYNEDSNKAKQETRAFISDYVLEMHPNENFENKLEEIIAENAVGNYTECIT
AAKLAIEKCGAKSVLLSFEPMNDLMSQKNVINIVDDNIKKYHMEYT"
gene 6605..7588
/gene="luxB"
/label=luxB
CDS 7695..8879
/codon_start=1
/gene="luxE"
/product="LuxE"
/label=luxE
/note="confers luminescent phenotype"
/protein_id="AGI04971.1"
/translation="MKGIKEYDSSAAILSNIILRSKTGMTSYVDKQEITASSEIDDLIF
SSDPLVWSYDEQEKIRKKLVLDAFRNHYKHCREYRHYCQAHKVDDNITEIDDIPVFPTS
VFKFTRLLTSQENEIESWFTSSGTNGLKSQVARDRLSIERLLGSVSYGMKYVGSWFDHQ
IELVNLGPDRFNAHNIWFKYVMSLVELLYPTTFTVTEERIDFVKTLNSLERIKNQGKDL
CLIGSPYFIYLLCHYMKDKKISFSGDKSLYIITGGGWKSYEKESLKRDDFNHLLFDTFN
LSDISQIRDIFNQVELNTCFFEDEMQRKHVPPWVYARALDPETLKPVPDGTPGLMSYMD
ASATSYPAFIVTDDVGIISREYGKYPGVLVEILRRVNTRTQKGCALSLTEAFDS"
gene 7695..8879
/gene="luxE"
/label=luxE
CDS 7767..8879
/codon_start=1
/gene="luxE"
/product="LuxE"
/label=LuxE
/note="from the Photorhabdus luminescens bioluminescence
operon"
/translation="MTSYVDKQEITASSEIDDLIFSSDPLVWSYDEQEKIRKKLVLDAF
RNHYKHCREYRHYCQAHKVDDNITEIDDIPVFPTSVFKFTRLLTSQENEIESWFTSSGT
NGLKSQVARDRLSIERLLGSVSYGMKYVGSWFDHQIELVNLGPDRFNAHNIWFKYVMSL
VELLYPTTFTVTEERIDFVKTLNSLERIKNQGKDLCLIGSPYFIYLLCHYMKDKKISFS
GDKSLYIITGGGWKSYEKESLKRDDFNHLLFDTFNLSDISQIRDIFNQVELNTCFFEDE
MQRKHVPPWVYARALDPETLKPVPDGTPGLMSYMDASATSYPAFIVTDDVGIISREYGK
YPGVLVEILRRVNTRTQKGCALSLTEAFDS"
misc_recomb 8914..8938
/label=attB2
/note="attB2"
protein_bind complement(8914..8938)
/gene="mutant version of attB"
/label=attB2
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
CDS 9368..10162
/codon_start=1
/gene="nptII"
/product="NptII"
/label=nptII
/note="kanamycin resistance protein"
/protein_id="AGI04972.1"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
gene 9368..10162
/gene="nptII"
/label=nptII
protein_bind complement(10261..10308)
/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)."
misc_recomb 10261..10307
/label=FRT
/note="FRT"
regulatory 10349..10446
/regulatory_class="terminator"
/note="T1"
terminator 10354..10440
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
regulatory 10543..10637
/regulatory_class="terminator"
/note="T0"
terminator 10543..10637
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
primer_bind complement(10665..10681)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 10686..10884
/label=Tn7R
/note="Tn7R"
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