Basic Vector Information
| Basic Vector Information | |||
|---|---|---|---|
| Vector Name | pTn5.7luxT5 | Length | 10774 bp |
| Type | Cloning vector | Source | Kvtiko BH, Bruckbauer ST, Karkhoff-Schweizer RR, Schweizer HP. |
pTn5.7luxT5 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 ℃.
pTn5.7luxT5 vector Sequence
LOCUS Exported 10774 bp ds-DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pTn5.7luxT5, complete sequence.
ACCESSION KF532959
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10774)
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JOURNAL Unpublished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 10774)
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 10774)
AUTHORS .
TITLE Direct Submission
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..10774
/organism="Cloning vector pTn5.7luxT5"
/mol_type="other DNA"
/db_xref="taxon:1424478"
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
gene complement(1446..3048)
/gene="tnpA"
/label=tnpA
CDS complement(1446..2888)
/codon_start=1
/gene="tnpA"
/product="TnpA"
/label=tnpA
/note="Tn5 transposase"
/protein_id="AHA61729.1"
/translation="MVTFMITSALHRAADWAKSVFSSAALGDPRHTARLVNVAAQLAKY
SGKSIPISSEGSEAAQEGAYRFIRNPNVSAEAIRKAGAMQTVKLAQEFPELLAIEDTTS
LSYRHQVAEELGKLGSIQDKSRGWWIHSVLLLEATTFRTVGLLHQEWWMRPDDPADADE
KESGKWLAAAATSRLRMGSMMSNVIAVCDREADIHAYLQDKLAHNERFVVRSKHPRKDV
ESGLYLYDHLKNQPELGGYQISIPQKGVVDKRGKRKNRPARKASLSLRSGRITLKQGNI
TLNAVLAEEINPPKGETPLKWLLLTSEPVESLAQALRVIDIYTHRWRIEEFHKAWKTGA
GAERQRMEEPDNLERMVSILSFVAVRLLQLRESFTLPQALRAQGLLKEAEHVESQSAET
VLTPDECQLLGYLDKGKRKRKEKAGSLQWAYMAIARLGGFMDSKRTGIASWGALWEGWE
ALQSKLDGFLAAKDLMAQGIKI"
regulatory complement(3019..3048)
/regulatory_class="promoter"
/gene="tnpA"
/note="S12 promoter"
misc_recomb 3055..3073
/label=Tn5 mosaic ends
/note="Tn5 mosaic ends"
misc_feature 3055..3073
/label=Tn5 ME
/note="hyperactive mosaic end for Tn5 transposase
recognition (Reznikoff et al., 2004)"
operon 3106..8903
/operon="luxCDABE operon"
/label=luxCDABE operon
CDS 3106..4548
/codon_start=1
/operon="luxCDABE operon"
/gene="luxC"
/product="LuxC"
/label=luxC
/note="confers luminescent phenotype"
/protein_id="AHA61731.1"
/translation="MTKKISFIINGQVEIFPESDDLVQSINFGDNSVYLPILNDSHVKN
IIDCNGNNELRLHNIVNFLYTVGQRWKNEEYSRRRTYIRDLKKYMGYSEEMAKLEANWI
SMILCSKGGLYDVVENELGSRHIMDEWLPQDESYVRAFPKGKSVHLLAGNVPLSGIMSI
LRAILTKNQCIIKTSSTDPFTANALALSFIDVDPNHPITRSLSVIYWPHQGDTSLAKEI
MRHADVIVAWGGPDAINWAVEHAPSYADVIKFGSKKSLCIIDNPVDLTSAATGAAHDVC
FYDQRACFSAQNIYYMGNHYEEFKLALIEKLNLYAHILPNAKKDFDEKAAYSLVQKESL
FAGLKVEVDIHQRWMIIESNAGVEFNQPLGRCVYLHHVDNIEQILPYVQKNKTQTISIF
PWESSFKYRDALALKGAERIVEAGMNNIFRVGGSHDGMRPLQRLVTYISHERPSNYTAK
DVAVEIEQTRFLEEDKFLVFVP"
gene 3106..4548
/operon="luxCDABE operon"
/gene="luxC"
/label=luxC
CDS 4560..5483
/codon_start=1
/operon="luxCDABE operon"
/gene="luxD"
/product="LuxD"
/label=luxD
/note="confers luminescent phenotype"
/protein_id="AHA61732.1"
/translation="MENESKYKTIDHVICVEGNKKIHVWETLPEENSPKRKNAIIIASG
FARRMDHFAGLAEYLSRNGFHVIRYDSLHHVGLSSGTIDEFTMSIGKQSLLAVVDWLTT
RKINNFGMLASSLSARIAYASLSEINASFLITAVGVVNLRYSLERALGFDYLSLPINEL
PDNLDFEGHKLGAEVFARDCLDFGWEDLASTINNMMYLDIPFIAFTANNDNWVKQDEVI
TLLSNIRSNRCKIYSLLGSSHDLSENLVVLRNFYQSVTKAAIAMDNDHLDIDVDITEPS
FEHLTIATVNERRMRIEIENQAISLS"
gene 4560..5483
/operon="luxCDABE operon"
/gene="luxD"
/label=luxD
CDS 5532..6614
/codon_start=1
/operon="luxCDABE operon"
/gene="luxA"
/product="LuxA"
/label=luxA
/note="confers luminescent phenotype"
/protein_id="AHA61733.1"
/translation="MKFGNFLLTYQPPQFSQTEVMKRLVKLGRISEECGFDTVWLLEHH
FTEFGLLGNPYVAAAYLLGATKKLNVGTAAIVLPTAHPVRQLEDVNLLDQMSKGRFRFG
ICRGLYNKDFRVFGTDMNNSRALAECWYGLIKNGMTEGYMEADNEHIKFHKVKVNPAAY
SRGGAPVYVVAESASTTEWAAQFGLPMILSWIINTNEKKAQLELYNEVAQEYGHDIHNI
DHCLSYITSVDHDSIKAKEICRKFLGHWYDSYVNATTIFDDSDQTRGYDFNKGQWRDFV
LKGHKDTNRRIDYSYEINPVGTPQECIDIIQKDIDATGISNICCGFEANGTVDEIIASM
KLFQSDVMPFLKEKQRSLLY"
gene 5532..6614
/operon="luxCDABE operon"
/gene="luxA"
/label=luxA
CDS 6629..7612
/codon_start=1
/operon="luxCDABE operon"
/gene="luxB"
/product="LuxB"
/label=luxB
/note="confers luminescent phenotype"
/protein_id="AHA61734.1"
/translation="MKFGLFFLNFINSTTVQEQSIVRMQEITEYVDKLNFEQILVYENH
FSDNGVVGAPLTVSGFLLGLTEKIKIGSLNHIITTHHPVAIAEEACLLDQLSEGRFILG
FSDCEKKDEMHFFNRPVEYQQQLFEECYEIINDALTTGYCNPDNDFYSFPKISVNPHAY
TPGGPRKYVTATSHHIVEWAAKKGIPLIFKWDDSNDVRYEYAERYKAVADKYDVDLSEI
DHQLMILVNYNEDSNKAKQETRAFISDYVLEMHPNENFENKLEEIIAENAVGNYTECIT
AAKLAIEKCGAKSVLLSFEPMNDLMSQKNVINIVDDNIKKYHMEYT"
gene 6629..7612
/operon="luxCDABE operon"
/gene="luxB"
/label=luxB
CDS 7719..8903
/codon_start=1
/operon="luxCDABE operon"
/gene="luxE"
/product="LuxE"
/label=luxE
/note="confers luminescent phenotype"
/protein_id="AHA61735.1"
/translation="MKGIKEYDSSAAILSNIILRSKTGMTSYVDKQEITASSEIDDLIF
SSDPLVWSYDEQEKIRKKLVLDAFRNHYKHCREYRHYCQAHKVDDNITEIDDIPVFPTS
VFKFTRLLTSQENEIESWFTSSGTNGLKSQVARDRLSIERLLGSVSYGMKYVGSWFDHQ
IELVNLGPDRFNAHNIWFKYVMSLVELLYPTTFTVTEERIDFVKTLNSLERIKNQGKDL
CLIGSPYFIYLLCHYMKDKKISFSGDKSLYIITGGGWKSYEKESLKRDDFNHLLFDTFN
LSDISQIRDIFNQVELNTCFFEDEMQRKHVPPWVYARALDPETLKPVPDGTPGLMSYMD
ASATSYPAFIVTDDVGIISREYGKYPGVLVEILRRVNTRTQKGCALSLTEAFDS"
gene 7719..8903
/operon="luxCDABE operon"
/gene="luxE"
/label=luxE
CDS 7791..8903
/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(8938..8962)
/gene="mutant version of attB"
/label=attB2
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
misc_recomb 9018..9065
/label=FRT
/note="FRT"
protein_bind complement(9018..9065)
/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 9117..9145
/gene="intI1 (promoter lies within the coding sequence)"
/label=Pc promoter
/note="class 1 integron promoter"
CDS 9472..9708
/codon_start=1
/gene="dhfrII"
/product="DhfrII"
/label=dhfrII
/note="trimethoprim resistance protein"
/protein_id="AHA61730.1"
/translation="MGQSSDEANAPVAGQFALPLSATFGLGDRVRKKSGAAWQGQVVGW
YCTKLTPEGYAVESESHPGSVQIYPVAALERVA"
gene 9472..9708
/gene="dhfrII"
/label=dhfrII
misc_recomb 9770..9817
/label=FRT
/note="FRT"
protein_bind complement(9770..9817)
/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 9858..9955
/regulatory_class="terminator"
/note="T1"
terminator 9863..9949
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
regulatory 10052..10146
/regulatory_class="terminator"
/note="T0"
terminator 10052..10146
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
primer_bind complement(10174..10190)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 10195..10393
/label=Tn7R
/note="Tn7R"
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