Basic Vector Information
| Basic Vector Information | |||
|---|---|---|---|
| Vector Name | pTn5.7luxTc6 | Length | 11969 bp |
| Type | Promoter capture vector | Source | Bruckbauer ST, Kvitko BH, Karkhoff-Schweizer RR, Schweizer HP. |
pTn5.7luxTc6 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 ℃.
pTn5.7luxTc6 vector Sequence
LOCUS Exported 11969 bp ds-DNA circular SYN 18-DEC-2018
DEFINITION Promoter capture vector pTn5.7luxTc6, complete sequence.
ACCESSION KC332288
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 11969)
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JOURNAL Unpublished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 11969)
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 11969)
AUTHORS .
TITLE Direct Submission
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..11969
/organism="Promoter capture vector pTn5.7luxTc6"
/mol_type="other DNA"
/db_xref="taxon:1303871"
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="AGI04980.1"
/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="AGI04981.1"
/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="AGI04982.1"
/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="AGI04983.1"
/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="AGI04984.1"
/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="AGI04985.1"
/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)."
CDS complement(9234..10424)
/codon_start=1
/gene="tetA"
/product="TetA"
/label=tetA
/note="tetracycline resistance protein"
/protein_id="AGI04986.1"
/translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
gene complement(9234..10424)
/gene="tetA"
/label=tetA
promoter complement(10472..10500)
/label=tet promoter
/note="E. coli promoter for tetracycline efflux protein
gene"
protein_bind 10747..10768
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 10783..10813
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 10821..10837
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
primer_bind 10845..10861
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
promoter 10879..10897
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
misc_recomb 10965..11012
/label=FRT
/note="FRT"
protein_bind complement(10965..11012)
/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 11053..11150
/regulatory_class="terminator"
/note="T1"
terminator 11058..11144
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
regulatory 11247..11341
/regulatory_class="terminator"
/note="T0"
terminator 11247..11341
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
primer_bind complement(11369..11385)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 11390..11588
/label=Tn7R
/note="Tn7R"
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