Basic Vector Information
| Basic Vector Information | |||
|---|---|---|---|
| Vector Name | pTn7oluxK4 | Length | 9823 bp |
| Type | Promoter capture vector | Source | Bruckbauer ST, Kvitko BH, Karkhoff-Schweizer RR, Schweizer HP. |
pTn7oluxK4 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 ℃.
pTn7oluxK4 vector Sequence
LOCUS Exported 9823 bp ds-DNA circular SYN 18-DEC-2018
DEFINITION Promoter capture vector pTn7oluxK4, complete sequence.
ACCESSION KC332284
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9823)
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JOURNAL Unpublished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 9823)
AUTHORS Bruckbauer ST, Kvitko BH, Karkhoff-Schweizer RR, Schweizer HP.
TITLE Direct Submission
JOURNAL Submitted (07-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 9823)
AUTHORS .
TITLE Direct Submission
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..9823
/organism="Promoter capture vector pTn7oluxK4"
/mol_type="other DNA"
/db_xref="taxon:1303867"
oriT 219..480
oriT 365..473
/note="incP origin of transfer"
rep_origin 504..885
/label=R6K gamma
/note="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)"
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"
regulatory 1350..1548
/regulatory_class="promoter"
/note="PompA region"
CDS 1581..3023
/codon_start=1
/gene="luxC"
/product="LuxC"
/label=luxC
/note="confers luminescent phenotype"
/protein_id="AGI04953.1"
/translation="MTKKISFIINGQVEIFPESDDLVQSINFGDNSVYLPILNDSHVKN
IIDCNGNNELRLHNIVNFLYTVGQRWKNEEYSRRRTYIRDLKKYMGYSEEMAKLEANWI
SMILCSKGGLYDVVENELGSRHIMDEWLPQDESYVRAFPKGKSVHLLAGNVPLSGIMSI
LRAILTKNQCIIKTSSTDPFTANALALSFIDVDPNHPITRSLSVIYWPHQGDTSLAKEI
MRHADVIVAWGGPDAINWAVEHAPSYADVIKFGSKKSLCIIDNPVDLTSAATGAAHDVC
FYDQRACFSAQNIYYMGNHYEEFKLALIEKLNLYAHILPNAKKDFDEKAAYSLVQKESL
FAGLKVEVDIHQRWMIIESNAGVEFNQPLGRCVYLHHVDNIEQILPYVQKNKTQTISIF
PWESSFKYRDALALKGAERIVEAGMNNIFRVGGSHDGMRPLQRLVTYISHERPSNYTAK
DVAVEIEQTRFLEEDKFLVFVP"
gene 1581..3023
/gene="luxC"
/label=luxC
CDS 3035..3958
/codon_start=1
/gene="luxD"
/product="LuxD"
/label=luxD
/note="confers luminescent phenotype"
/protein_id="AGI04954.1"
/translation="MENESKYKTIDHVICVEGNKKIHVWETLPEENSPKRKNAIIIASG
FARRMDHFAGLAEYLSRNGFHVIRYDSLHHVGLSSGTIDEFTMSIGKQSLLAVVDWLTT
RKINNFGMLASSLSARIAYASLSEINASFLITAVGVVNLRYSLERALGFDYLSLPINEL
PDNLDFEGHKLGAEVFARDCLDFGWEDLASTINNMMYLDIPFIAFTANNDNWVKQDEVI
TLLSNIRSNRCKIYSLLGSSHDLSENLVVLRNFYQSVTKAAIAMDNDHLDIDVDITEPS
FEHLTIATVNERRMRIEIENQAISLS"
gene 3035..3958
/gene="luxD"
/label=luxD
CDS 4007..5089
/codon_start=1
/gene="luxA"
/product="LuxA"
/label=luxA
/note="confers luminescent phenotype"
/protein_id="AGI04955.1"
/translation="MKFGNFLLTYQPPQFSQTEVMKRLVKLGRISEECGFDTVWLLEHH
FTEFGLLGNPYVAAAYLLGATKKLNVGTAAIVLPTAHPVRQLEDVNLLDQMSKGRFRFG
ICRGLYNKDFRVFGTDMNNSRALAECWYGLIKNGMTEGYMEADNEHIKFHKVKVNPAAY
SRGGAPVYVVAESASTTEWAAQFGLPMILSWIINTNEKKAQLELYNEVAQEYGHDIHNI
DHCLSYITSVDHDSIKAKEICRKFLGHWYDSYVNATTIFDDSDQTRGYDFNKGQWRDFV
LKGHKDTNRRIDYSYEINPVGTPQECIDIIQKDIDATGISNICCGFEANGTVDEIIASM
KLFQSDVMPFLKEKQRSLLY"
gene 4007..5089
/gene="luxA"
/label=luxA
CDS 5104..6087
/codon_start=1
/gene="luxB"
/product="LuxB"
/label=luxB
/note="confers luminescent phenotype"
/protein_id="AGI04956.1"
/translation="MKFGLFFLNFINSTTVQEQSIVRMQEITEYVDKLNFEQILVYENH
FSDNGVVGAPLTVSGFLLGLTEKIKIGSLNHIITTHHPVAIAEEACLLDQLSEGRFILG
FSDCEKKDEMHFFNRPVEYQQQLFEECYEIINDALTTGYCNPDNDFYSFPKISVNPHAY
TPGGPRKYVTATSHHIVEWAAKKGIPLIFKWDDSNDVRYEYAERYKAVADKYDVDLSEI
DHQLMILVNYNEDSNKAKQETRAFISDYVLEMHPNENFENKLEEIIAENAVGNYTECIT
AAKLAIEKCGAKSVLLSFEPMNDLMSQKNVINIVDDNIKKYHMEYT"
gene 5104..6087
/gene="luxB"
/label=luxB
CDS 6194..7378
/codon_start=1
/gene="luxE"
/product="LuxE"
/label=luxE
/note="confers luminescent phenotype"
/protein_id="AGI04957.1"
/translation="MKGIKEYDSSAAILSNIILRSKTGMTSYVDKQEITASSEIDDLIF
SSDPLVWSYDEQEKIRKKLVLDAFRNHYKHCREYRHYCQAHKVDDNITEIDDIPVFPTS
VFKFTRLLTSQENEIESWFTSSGTNGLKSQVARDRLSIERLLGSVSYGMKYVGSWFDHQ
IELVNLGPDRFNAHNIWFKYVMSLVELLYPTTFTVTEERIDFVKTLNSLERIKNQGKDL
CLIGSPYFIYLLCHYMKDKKISFSGDKSLYIITGGGWKSYEKESLKRDDFNHLLFDTFN
LSDISQIRDIFNQVELNTCFFEDEMQRKHVPPWVYARALDPETLKPVPDGTPGLMSYMD
ASATSYPAFIVTDDVGIISREYGKYPGVLVEILRRVNTRTQKGCALSLTEAFDS"
gene 6194..7378
/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"
misc_recomb 7413..7437
/label=attB2
/note="attB2"
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)."
CDS 7926..8720
/codon_start=1
/gene="nptII"
/product="NptII"
/label=nptII
/note="kanamycin resistance protein"
/protein_id="AGI04958.1"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
gene 7926..8720
/gene="nptII"
/label=nptII
misc_recomb 8819..8866
/label=FRT
/note="FRT"
protein_bind complement(8819..8866)
/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 8907..9004
/regulatory_class="terminator"
/note="T1"
terminator 8912..8998
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
regulatory 9101..9195
/regulatory_class="terminator"
/note="T0"
terminator 9101..9195
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
primer_bind complement(9223..9239)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 9244..9442
/label=Tn7R
/note="Tn7R"
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