Cart 2

pTn5.7luxK4 vector (V002599#)

Basic Vector Information

Basic Vector Information
Vector Name pTn5.7luxK4 Length 11324 bp
Type Cloning vector Source Kvtiko BH, Bruckbauer ST, Karkhoff-Schweizer RR, Schweizer HP.

pTn5.7luxK4 vector Vector Map

oriT R6K oriV Tn7L Tn5 mosaic ends AmpR promoter tnpA Tn5 mosaic ends luxCDABE operon attB2 FRT nptII FRT T0 T1 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 ℃.

pTn5.7luxK4 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported               11324 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pTn5.7luxK4, complete sequence.
ACCESSION   KF532957
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  JOURNAL   Unpublished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)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..11324
                     /organism="Cloning vector pTn5.7luxK4"
                     /mol_type="other DNA"
                     /db_xref="taxon:1424477"
     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)"
     misc_recomb     1276..1294
                     /label=Tn5 mosaic ends
                     /note="Tn5 mosaic ends"
     misc_feature    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(1446..2888)
                     /codon_start=1
                     /gene="tnpA"
                     /product="TnpA"
                     /label=tnpA
                     /note="Tn5 transposase"
                     /protein_id="AHA61715.1"
                     /translation="MVTFMITSALHRAADWAKSVFSSAALGDPRRTARLVNVAAQLAKY
                     SGKSIPISSEGSEAAQEGAYRFIRNPNVSAEAIRKAGAMQTVKLAQEFPELLAIEDTTS
                     LSYRHQVAEELGKLGSIQDKSRGWWVHSVLLLEATTFRTVGLLHQEWWMRPDDPADADE
                     KESGKWLAAAATSRLRMGSMMSNVIAVCDREADIHAYLQDKLAHNERFVVRSKHPRKDV
                     ESGLYLYDHLKNQPELGGYQISIPQKGVVDKRGKRKNRPARKASLSLRSGRITLKQGNI
                     TLNAVLAEEINPPKGETPLKWLLLTSEPVESLAQALRVIDIYTHRWRIEEFHKAWKTGA
                     GAERQRMEEPDNLERMVSILSFVAVRLLQLRESFTLPQALRAQGLLKEAEHVESQSAET
                     VLTPDECQLLGYLDKGKRKRKEKAGSLQWAYMAIARLGGFMDSKRTGIASWGALWEGWE
                     ALQSKLDGFLAAKDLMAQGIKI"
     gene            complement(1446..2888)
                     /gene="tnpA"
                     /label=tnpA
     misc_recomb     3031..3049
                     /label=Tn5 mosaic ends
                     /note="Tn5 mosaic ends"
     misc_feature    3031..3049
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase 
                     recognition (Reznikoff et al., 2004)"
     operon          3082..8879
                     /operon="luxCDABE operon"
                     /label=luxCDABE operon
     CDS             3082..4524
                     /codon_start=1
                     /operon="luxCDABE operon"
                     /gene="luxC"
                     /product="LuxC"
                     /label=luxC
                     /note="confers luminescent phenotype"
                     /protein_id="AHA61717.1"
                     /translation="MTKKISFIINGQVEIFPESDDLVQSINFGDNSVYLPILNDSHVKN
                     IIDCNGNNELRLHNIVNFLYTVGQRWKNEEYSRRRTYIRDLKKYMGYSEEMAKLEANWI
                     SMILCSKGGLYDVVENELGSRHIMDEWLPQDESYVRAFPKGKSVHLLAGNVPLSGIMSI
                     LRAILTKNQCIIKTSSTDPFTANALALSFIDVDPNHPITRSLSVIYWPHQGDTSLAKEI
                     MRHADVIVAWGGPDAINWAVEHAPSYADVIKFGSKKSLCIIDNPVDLTSAATGAAHDVC
                     FYDQRACFSAQNIYYMGNHYEEFKLALIEKLNLYAHILPNAKKDFDEKAAYSLVQKESL
                     FAGLKVEVDIHQRWMIIESNAGVEFNQPLGRCVYLHHVDNIEQILPYVQKNKTQTISIF
                     PWESSFKYRDALALKGAERIVEAGMNNIFRVGGSHDGMRPLQRLVTYISHERPSNYTAK
                     DVAVEIEQTRFLEEDKFLVFVP"
     gene            3082..4524
                     /operon="luxCDABE operon"
                     /gene="luxC"
                     /label=luxC
     CDS             4536..5459
                     /codon_start=1
                     /operon="luxCDABE operon"
                     /gene="luxD"
                     /product="LuxD"
                     /label=luxD
                     /note="confers luminescent phenotype"
                     /protein_id="AHA61718.1"
                     /translation="MENESKYKTIDHVICVEGNKKIHVWETLPEENSPKRKNAIIIASG
                     FARRMDHFAGLAEYLSRNGFHVIRYDSLHHVGLSSGTIDEFTMSIGKQSLLAVVDWLTT
                     RKINNFGMLASSLSARIAYASLSEINASFLITAVGVVNLRYSLERALGFDYLSLPINEL
                     PDNLDFEGHKLGAEVFARDCLDFGWEDLASTINNMMYLDIPFIAFTANNDNWVKQDEVI
                     TLLSNIRSNRCKIYSLLGSSHDLSENLVVLRNFYQSVTKAAIAMDNDHLDIDVDITEPS
                     FEHLTIATVNERRMRIEIENQAISLS"
     gene            4536..5459
                     /operon="luxCDABE operon"
                     /gene="luxD"
                     /label=luxD
     CDS             5508..6590
                     /codon_start=1
                     /operon="luxCDABE operon"
                     /gene="luxA"
                     /product="LuxA"
                     /label=luxA
                     /note="confers luminescent phenotype"
                     /protein_id="AHA61719.1"
                     /translation="MKFGNFLLTYQPPQFSQTEVMKRLVKLGRISEECGFDTVWLLEHH
                     FTEFGLLGNPYVAAAYLLGATKKLNVGTAAIVLPTAHPVRQLEDVNLLDQMSKGRFRFG
                     ICRGLYNKDFRVFGTDMNNSRALAECWYGLIKNGMTEGYMEADNEHIKFHKVKVNPAAY
                     SRGGAPVYVVAESASTTEWAAQFGLPMILSWIINTNEKKAQLELYNEVAQEYGHDIHNI
                     DHCLSYITSVDHDSIKAKEICRKFLGHWYDSYVNATTIFDDSDQTRGYDFNKGQWRDFV
                     LKGHKDTNRRIDYSYEINPVGTPQECIDIIQKDIDATGISNICCGFEANGTVDEIIASM
                     KLFQSDVMPFLKEKQRSLLY"
     gene            5508..6590
                     /operon="luxCDABE operon"
                     /gene="luxA"
                     /label=luxA
     CDS             6605..7588
                     /codon_start=1
                     /operon="luxCDABE operon"
                     /gene="luxB"
                     /product="LuxB"
                     /label=luxB
                     /note="confers luminescent phenotype"
                     /protein_id="AHA61720.1"
                     /translation="MKFGLFFLNFINSTTVQEQSIVRMQEITEYVDKLNFEQILVYENH
                     FSDNGVVGAPLTVSGFLLGLTEKIKIGSLNHIITTHHPVAIAEEACLLDQLSEGRFILG
                     FSDCEKKDEMHFFNRPVEYQQQLFEECYEIINDALTTGYCNPDNDFYSFPKISVNPHAY
                     TPGGPRKYVTATSHHIVEWAAKKGIPLIFKWDDSNDVRYEYAERYKAVADKYDVDLSEI
                     DHQLMILVNYNEDSNKAKQETRAFISDYVLEMHPNENFENKLEEIIAENAVGNYTECIT
                     AAKLAIEKCGAKSVLLSFEPMNDLMSQKNVINIVDDNIKKYHMEYT"
     gene            6605..7588
                     /operon="luxCDABE operon"
                     /gene="luxB"
                     /label=luxB
     CDS             7695..8879
                     /codon_start=1
                     /operon="luxCDABE operon"
                     /gene="luxE"
                     /product="LuxE"
                     /label=luxE
                     /note="confers luminescent phenotype"
                     /protein_id="AHA61721.1"
                     /translation="MKGIKEYDSSAAILSNIILRSKTGMTSYVDKQEITASSEIDDLIF
                     SSDPLVWSYDEQEKIRKKLVLDAFRNHYKHCREYRHYCQAHKVDDNITEIDDIPVFPTS
                     VFKFTRLLTSQENEIESWFTSSGTNGLKSQVARDRLSIERLLGSVSYGMKYVGSWFDHQ
                     IELVNLGPDRFNAHNIWFKYVMSLVELLYPTTFTVTEERIDFVKTLNSLERIKNQGKDL
                     CLIGSPYFIYLLCHYMKDKKISFSGDKSLYIITGGGWKSYEKESLKRDDFNHLLFDTFN
                     LSDISQIRDIFNQVELNTCFFEDEMQRKHVPPWVYARALDPETLKPVPDGTPGLMSYMD
                     ASATSYPAFIVTDDVGIISREYGKYPGVLVEILRRVNTRTQKGCALSLTEAFDS"
     gene            7695..8879
                     /operon="luxCDABE operon"
                     /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"
     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"
     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..10221
                     /codon_start=1
                     /gene="nptII"
                     /product="NptII"
                     /label=nptII
                     /note="kanamycin resistance protein"
                     /protein_id="AHA61716.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     gene            9427..10221
                     /gene="nptII"
                     /label=nptII
                     /note="kanamycin resistance gene"
     misc_recomb     10320..10367
                     /label=FRT
                     /note="FRT"
     protein_bind    complement(10320..10367)
                     /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      10408..10505
                     /regulatory_class="terminator"
                     /note="T0"
     terminator      10413..10499
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     regulatory      10602..10696
                     /regulatory_class="terminator"
                     /note="T1"
     terminator      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"

This page is informational only.