Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V000958 | pBAV1k-T5-LUX | In stock, 1 week for quality controls |
Buy one, get one free! |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
- Vector Name:
- pBAV1k-T5-LUX
- Antibiotic Resistance:
- Kanamycin
- Length:
- 8599 bp
- Type:
- Synthetic Biology
- Copy Number:
- Low Copy
- Promoter:
- T5
- Cloning Method:
- Restriction Enzyme
pBAV1k-T5-LUX vector Map
Plasmid 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 ℃.
6. The concentration of plasmid re-measurement sometimes differs from the nominal value, which may be due to the position of the lyophilized plasmid in the tube, the efficiency of the re-dissolution, the measurement bias, and adsorption on the wall of the tube, therefore, it is recommended to transform and extract the plasmid before using it
General Plasmid Transform Protocol
1. Take one 100μl of the competent cells and thaw it on ice for 10min, add 2μl of plasmid, then ice bath for 30min, then heat-shock it at 42℃ for 60s, do not stir, and then ice bath for 2min.
2. Add 900μl of LB liquid medium without antibiotics, and incubate at 37℃ for 45min (30℃ for 1-1.5 hours) with 180rpm shaking.
3. Centrifuge at 6000rpm for 5min, leave only 100μl of supernatant to resuspend the bacterial precipitate and spread it onto the target plasmid-resistant LB plate.
4. Invert the plate and incubate at 37℃ for 14h, or at 30℃ for 20h.
5. Pick a single colony into LB liquid medium, add the corresponding antibiotics, incubate at 220rpm for 14h, and extract the plasmid according to the experimental needs and the instructions of the plasmid extraction kit.
pBAV1k-T5-LUX vector Sequence
LOCUS Exported 8599 bp ds-DNA circular SYN 13-MAY-2021
DEFINITION Expresses the Photorhabdus luminescens lux operon in bacterial
cells.
ACCESSION .
VERSION .
KEYWORDS pBAV1k-T5-LUX
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8599)
AUTHORS Bryksin AV, Matsumura I
TITLE Rational design of a plasmid origin that replicates efficiently in
both gram-positive and gram-negative bacteria.
JOURNAL PLoS One. 2010 . 5(10):e13244.
PUBMED 20949038
REFERENCE 2 (bases 1 to 8599)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "PLoS One.
2010 . 5(10):e13244."
FEATURES Location/Qualifiers
source 1..8599
/organism="synthetic DNA construct"
/mol_type="other DNA"
protein_bind 14..30
/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)."
RBS 40..51
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
CDS 58..1140
/codon_start=1
/gene="luxA"
/product="LuxA luciferase subunit"
/label=LuxA
/note="from the Photorhabdus luminescens bioluminescence
operon"
/translation="MKFGNFLLTYQPPQFSQTEVMKRLVKLGRISEECGFDTVWLLEHH
FTEFGLLGNPYVAAAYLLGATKKLNVGTAAIVLPTAHPVRQLEDVNLLDQMSKGRFRFG
ICRGLYNKDFRVFGTDMNNSRALAECWYGLIKNGMTEGYMEADNEHIKFHKVKVNPAAY
SRGGAPVYVVAESASTTEWAAQFGLPMILSWIINTNEKKAQLELYNEVAQEYGHDIHNI
DHCLSYITSVDHDSIKAKEICRKFLGHWYDSYVNATTIFDDSDQTRGYDFNKGQWRDFV
LKGHKDTNRRIDYSYEINPVGTPQECIDIIQKDIDATGISNICCGFEANGTVDEIIASM
KLFQSDVMPFLKEKQRSLLY"
RBS 1152..1163
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
CDS 1170..2153
/codon_start=1
/gene="luxB"
/product="LuxB luciferase subunit"
/label=LuxB
/note="from the Photorhabdus luminescens bioluminescence
operon"
/translation="MKFGLFFLNFINSTTVQEQSIVRMQEITEYVDKLNFEQILVYENH
FSDNGVVGAPLTVSGFLLGLTEKIKIGSLNHIITTHHPVRIAEEACLLDQLSEGRFILG
FSDCEKKDEMHFFNRPVEYQQQLFEECYEIINDALTTGYCNPDNDFYSFPKISVNPHAY
TPGGPRKYVTATSHHIVEWAAKKGIPLIFKWDDSNDVRYEYAERYKAVADKYDVDLSEI
DHQLMILVNYNEDSNKAKQETRAFISDYVLEMHPNENFENKLEEIIAENAVGNYTECIT
AAKLAIEKCGAKSVLLSFEPMNDLMSQKNVINIVDDNIKKYHMEYT"
RBS 2165..2176
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
CDS 2183..3625
/codon_start=1
/gene="luxC"
/product="LuxC fatty acid reductase"
/label=LuxC
/note="from the Photorhabdus luminescens bioluminescence
operon"
/translation="MTKKISFIINGQVEIFPESDDLVQSINFGDNSVYLPILNDSHVKN
IIDCNGNNELRLHNIVNFLYTVGQRWKNEEYSRRRTYIRDLKKYMGYSEEMAKLEANWI
SMILCSKGGLYDVVENELGSRHIMDEWLPQDESYVRAFPKGKSVHLLAGNVPLSGIMSI
LRAILTKNQCIIKTSSTDPFTANALALSFIDVDPNHPITRSLSVIYWPHQGDTSLAKEI
MRHADVIVAWGGPDAINWAVEHAPSYADVIKFGSKKSLCIIDNPVDLTSAATGAAHDVC
FYDQRACFSAQNIYYMGNHYEEFKLALIEKLNLYAHILPNAKKDFDEKAAYSLVQKESL
FAGLKVEVDIHQRWMIIESNAGVEFNQPLGRCVYLHHVDNIEQILPYVQKNKTQTISIF
PWESSFKYRDALALKGAERIVEAGMNNIFRVGGSHDGMRPLQRLVTYISHERPSNYTAK
DVAVEIEQTRFLEEDKFLVFVP"
RBS 3637..3648
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
CDS 3655..4578
/codon_start=1
/gene="luxD"
/product="LuxD acyltransferase"
/label=LuxD
/note="from the Photorhabdus luminescens bioluminescence
operon"
/translation="MENESKYKTIDHVICVEGNKKIHVWETLPEENSPKRKNAIIIASG
FARRMDHFAGLAEYLSRNGFHVIRYDSLHHVGLSSGTIDEFTMSIGKQSLLAVVDWLTT
RKINNFGMLASSLSARIAYASLSEINASFLITAVGVVNLRYSLERALGFDYLSLPINEL
PDNLDFEGHKLGAEVFARDCLDFGWEDLASTINNMMYLDIPFIAFTANNDNWVKQDEVI
TLLSNIRSNRCKIYSLLGSSHDLSENLVVLRNFYQSVTKAAIAMDNDHLDIDVDITEPS
FEHLTIATVNERRMRIEIENQAISLS"
RBS 4590..4601
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
CDS 4608..5720
/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_feature 5724..5744
/label=BioBrick suffix
/note="universal suffix for all parts"
terminator 5795..5881
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
CDS 7212..7223
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
terminator 7425..7519
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
CDS complement(7535..8329)
/codon_start=1
/gene="aphA-3"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin"
/translation="MAKMRISPELKKLIEEYRCVKDTEGMSPAKVYKLVGENEILYLKM
TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEGYED
EQSPEKTIELYAECIRLFHSIDISDCPYTNSLDGRLAELDYLLNNDLADVDCENWEEDT
PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
misc_feature 8535..8556
/label=BioBrick prefix
/note="BioBrick prefix for parts that do not start with
""ATG"""
promoter join(8561..8599,1..6)
/label=T5 promoter
/note="bacteriophage T5 promoter for E. coli RNA
polymerase, with embedded lac operator"
protein_bind 8581..8597
/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)."