| 载体基本信息 |
| 载体名称 |
pBAV1k-T5-LUX |
载体抗性 |
Kanamycin |
| 载体长度 |
8599 bp |
载体类型 |
Synthetic Biology |
| 拷贝数 |
Low Copy |
启动子 |
T5 |
| 克隆方法 |
Restriction Enzyme |
pBAV1k-T5-LUX 载体载体图谱
pBAV1k-T5-LUX 载体载体序列
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)."