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pTn5.7luxT6 vector (V002595#)

Basic Vector Information

Basic Vector Information
Vector Name pTn5.7luxT6 Length 10805 bp
Type Promoter capture vector Source Bruckbauer ST, Kvitko BH, Karkhoff-Schweizer RR, Schweizer HP.

pTn5.7luxT6 vector Vector Map

R6K gamma Tn7L Tn5 mosaic end AmpR promoter tnpA PS12 Tn5 mosaic end attB1 luxC luxD luxA luxB luxE attB2 FRT Pc promoter dhfrII 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 ℃.

pTn5.7luxT6 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported               10805 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Promoter capture vector pTn5.7luxT6, complete sequence.
ACCESSION   KC332285
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10805)
  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  JOURNAL   Unpublished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 10805)
  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 10805)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..10805
                     /organism="Promoter capture vector pTn5.7luxT6"
                     /mol_type="other DNA"
                     /db_xref="taxon:1303868"
     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)"
     misc_feature    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="AGI04959.1"
                     /translation="MVTFMITSALHRAADWAKSVFSSAALGDPRHTARLVNVAAQLAKY
                     SGKSIPISSEGSEAAQEGAYRFIRNPNVSAEAIRKAGAMQTVKLAQEFPELLAIEDTTS
                     LSYRHQVAEELGKLGSIQDKSRGWWIHSVLLLEATTFRTVGLLHQEWWMRPDDPADADE
                     KESGKWLAAAATSRLRMGSMMSNVIAVCDREADIHAYLQDKLAHNERFVVRSKHPRKDV
                     ESGLYLYDHLKNQPELGGYQISIPQKGVVDKRGKRKNRPARKASLSLRSGRITLKQGNI
                     TLNAVLAEEINPPKGETPLKWLLLTSEPVESLAQALRVIDIYTHRWRIEEFHKAWKTGA
                     GAERQRMEEPDNLERMVSILSFVAVRLLQLRESFTLPQALRAQGLLKEAEHVESQSAET
                     VLTPDECQLLGYLDKGKRKRKEKAGSLQWAYMAIARLGGFMDSKRTGIASWGALWEGWE
                     ALQSKLDGFLAAKDLMAQGIKI"
     gene            complement(1446..2888)
                     /gene="tnpA"
                     /label=tnpA
     regulatory      complement(3019..3048)
                     /regulatory_class="promoter"
                     /note="PS12"
     misc_feature    3055..3073
                     /label=Tn5 mosaic end
                     /note="Tn5 mosaic end"
     misc_recomb     3083..3107
                     /label=attB1
                     /note="attB1"
     protein_bind    3083..3107
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3137..4579
                     /codon_start=1
                     /gene="luxC"
                     /product="LuxC"
                     /label=luxC
                     /note="confers luminescent phenotype"
                     /protein_id="AGI04960.1"
                     /translation="MTKKISFIINGQVEIFPESDDLVQSINFGDNSVYLPILNDSHVKN
                     IIDCNGNNELRLHNIVNFLYTVGQRWKNEEYSRRRTYIRDLKKYMGYSEEMAKLEANWI
                     SMILCSKGGLYDVVENELGSRHIMDEWLPQDESYVRAFPKGKSVHLLAGNVPLSGIMSI
                     LRAILTKNQCIIKTSSTDPFTANALALSFIDVDPNHPITRSLSVIYWPHQGDTSLAKEI
                     MRHADVIVAWGGPDAINWAVEHAPSYADVIKFGSKKSLCIIDNPVDLTSAATGAAHDVC
                     FYDQRACFSAQNIYYMGNHYEEFKLALIEKLNLYAHILPNAKKDFDEKAAYSLVQKESL
                     FAGLKVEVDIHQRWMIIESNAGVEFNQPLGRCVYLHHVDNIEQILPYVQKNKTQTISIF
                     PWESSFKYRDALALKGAERIVEAGMNNIFRVGGSHDGMRPLQRLVTYISHERPSNYTAK
                     DVAVEIEQTRFLEEDKFLVFVP"
     gene            3137..4579
                     /gene="luxC"
                     /label=luxC
     CDS             4591..5514
                     /codon_start=1
                     /gene="luxD"
                     /product="LuxD"
                     /label=luxD
                     /note="confers luminescent phenotype"
                     /protein_id="AGI04961.1"
                     /translation="MENESKYKTIDHVICVEGNKKIHVWETLPEENSPKRKNAIIIASG
                     FARRMDHFAGLAEYLSRNGFHVIRYDSLHHVGLSSGTIDEFTMSIGKQSLLAVVDWLTT
                     RKINNFGMLASSLSARIAYASLSEINASFLITAVGVVNLRYSLERALGFDYLSLPINEL
                     PDNLDFEGHKLGAEVFARDCLDFGWEDLASTINNMMYLDIPFIAFTANNDNWVKQDEVI
                     TLLSNIRSNRCKIYSLLGSSHDLSENLVVLRNFYQSVTKAAIAMDNDHLDIDVDITEPS
                     FEHLTIATVNERRMRIEIENQAISLS"
     gene            4591..5514
                     /gene="luxD"
                     /label=luxD
     CDS             5563..6645
                     /codon_start=1
                     /gene="luxA"
                     /product="LuxA"
                     /label=luxA
                     /note="confers luminescent phenotype"
                     /protein_id="AGI04962.1"
                     /translation="MKFGNFLLTYQPPQFSQTEVMKRLVKLGRISEECGFDTVWLLEHH
                     FTEFGLLGNPYVAAAYLLGATKKLNVGTAAIVLPTAHPVRQLEDVNLLDQMSKGRFRFG
                     ICRGLYNKDFRVFGTDMNNSRALAECWYGLIKNGMTEGYMEADNEHIKFHKVKVNPAAY
                     SRGGAPVYVVAESASTTEWAAQFGLPMILSWIINTNEKKAQLELYNEVAQEYGHDIHNI
                     DHCLSYITSVDHDSIKAKEICRKFLGHWYDSYVNATTIFDDSDQTRGYDFNKGQWRDFV
                     LKGHKDTNRRIDYSYEINPVGTPQECIDIIQKDIDATGISNICCGFEANGTVDEIIASM
                     KLFQSDVMPFLKEKQRSLLY"
     gene            5563..6645
                     /gene="luxA"
                     /label=luxA
     CDS             6660..7643
                     /codon_start=1
                     /gene="luxB"
                     /product="LuxB"
                     /label=luxB
                     /note="confers luminescent phenotype"
                     /protein_id="AGI04963.1"
                     /translation="MKFGLFFLNFINSTTVQEQSIVRMQEITEYVDKLNFEQILVYENH
                     FSDNGVVGAPLTVSGFLLGLTEKIKIGSLNHIITTHHPVAIAEEACLLDQLSEGRFILG
                     FSDCEKKDEMHFFNRPVEYQQQLFEECYEIINDALTTGYCNPDNDFYSFPKISVNPHAY
                     TPGGPRKYVTATSHHIVEWAAKKGIPLIFKWDDSNDVRYEYAERYKAVADKYDVDLSEI
                     DHQLMILVNYNEDSNKAKQETRAFISDYVLEMHPNENFENKLEEIIAENAVGNYTECIT
                     AAKLAIEKCGAKSVLLSFEPMNDLMSQKNVINIVDDNIKKYHMEYT"
     gene            6660..7643
                     /gene="luxB"
                     /label=luxB
     CDS             7750..8934
                     /codon_start=1
                     /gene="luxE"
                     /product="LuxE"
                     /label=luxE
                     /note="confers luminescent phenotype"
                     /protein_id="AGI04964.1"
                     /translation="MKGIKEYDSSAAILSNIILRSKTGMTSYVDKQEITASSEIDDLIF
                     SSDPLVWSYDEQEKIRKKLVLDAFRNHYKHCREYRHYCQAHKVDDNITEIDDIPVFPTS
                     VFKFTRLLTSQENEIESWFTSSGTNGLKSQVARDRLSIERLLGSVSYGMKYVGSWFDHQ
                     IELVNLGPDRFNAHNIWFKYVMSLVELLYPTTFTVTEERIDFVKTLNSLERIKNQGKDL
                     CLIGSPYFIYLLCHYMKDKKISFSGDKSLYIITGGGWKSYEKESLKRDDFNHLLFDTFN
                     LSDISQIRDIFNQVELNTCFFEDEMQRKHVPPWVYARALDPETLKPVPDGTPGLMSYMD
                     ASATSYPAFIVTDDVGIISREYGKYPGVLVEILRRVNTRTQKGCALSLTEAFDS"
     gene            7750..8934
                     /gene="luxE"
                     /label=luxE
     CDS             7822..8934
                     /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     8969..8993
                     /label=attB2
                     /note="attB2"
     protein_bind    complement(8969..8993)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_recomb     9049..9096
                     /label=FRT
                     /note="FRT"
     protein_bind    complement(9049..9096)
                     /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)."
     promoter        9148..9176
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             9503..9739
                     /codon_start=1
                     /gene="dhfrII"
                     /product="DhfrII"
                     /label=dhfrII
                     /note="trimethoprim resistance protein"
                     /protein_id="AGI04965.1"
                     /translation="MGQSSDEANAPVAGQFALPLSATFGLGDRVRKKSGAAWQGQVVGW
                     YCTKLTPEGYAVESESHPGSVQIYPVAALERVA"
     gene            9503..9739
                     /gene="dhfrII"
                     /label=dhfrII
     misc_recomb     9801..9848
                     /label=FRT
                     /note="FRT"
     protein_bind    complement(9801..9848)
                     /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      9889..9986
                     /regulatory_class="terminator"
                     /note="T1"
     terminator      9894..9980
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     regulatory      10083..10177
                     /regulatory_class="terminator"
                     /note="T0"
     terminator      10083..10177
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(10205..10221)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    10226..10424
                     /label=Tn7R
                     /note="Tn7R"

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