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pTn5.7luxK3 vector (V002600#)

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

Tn7L Tn5 mosaic end AmpR promoter tnpA Tn5 mosaic end luxC luxD luxA luxB luxE attB2 nptII FRT T1 T0 KS primer Tn7R

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

Copy Sequence

Download GeneBank File(.gb)

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  JOURNAL   Unpublished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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