- ￥100 - 1000
- Xybio
- 上海
- P0915
- 2025年07月11日
9 年钻石会员
企业认证
相关产品推荐更多 >
万千商家帮你免费找货
0 人在求购买到急需产品
- 详细信息
- 询价记录
- 文献和实验
- 技术资料
- 保存条件：
-20
- 保质期：
2年
- 英文名：
Plasmid#55800;LuxCDABE
- 库存：
100
- 供应商：
上海烜雅生物科技有限公司
- 规格：
干粉/液体
基本信息
名称：pBAV1k-T5-LUX基因质粒
别称: Plasmid#55800;LuxCDABE
启动子：T5
复制子: pUC
终止子: rnB T1 terminator
质粒分类: 基因文库质粒；其它基因质粒；其它种源质粒
质粒大小：8599bp
原核抗性: Kan
筛选标记：G418
克隆菌株: DH5a
培养条件: 30度
备注: 低拷贝
质粒属性
载体宿主：大肠杆菌
载体用途：蛋白表达
基因种属：
基因类型：ORF
原核抗性：Kan
真核抗性：
荧光蛋白：
质粒简介
Expresses the Photorhabdus luminescens lux operon in bacterial cells.The copy number of pBAV1k plasmids are not well controlled, so purification yields vary. A lacI+ E. coli strain will prevent constitutive expression of the lux operon.
质粒图谱
质粒序列
LOCUS Exported 8599 bp ds-DNA circular SYN 15-AUG-2017
DEFINITION synthetic circular DNA
FEATURES Location/Qualifiers
source 1..8599
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 26..70
/label=T5 promoter
/note="bacteriophage T5 promoter for E. coli RNA
polymerase, with embedded lac operator"
protein_bind 46..62
/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)."
protein_bind 78..94
/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 104..115
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
CDS 122..1204
/codon_start=1
/gene="luxA"
/product="LuxA luciferase subunit"
/note="from the Photorhabdus luminescens bioluminescence
operon"
/translation="MKFGNFLLTYQPPQFSQTEVMKRLVKLGRISEECGFDTVWLLEHH
FTEFGLLGNPYVAAAYLLGATKKLNVGTAAIVLPTAHPVRQLEDVNLLDQMSKGRFRFG
ICRGLYNKDFRVFGTDMNNSRALAECWYGLIKNGMTEGYMEADNEHIKFHKVKVNPAAY
SRGGAPVYVVAESASTTEWAAQFGLPMILSWIINTNEKKAQLELYNEVAQEYGHDIHNI
DHCLSYITSVDHDSIKAKEICRKFLGHWYDSYVNATTIFDDSDQTRGYDFNKGQWRDFV
LKGHKDTNRRIDYSYEINPVGTPQECIDIIQKDIDATGISNICCGFEANGTVDEIIASM
KLFQSDVMPFLKEKQRSLLY"
RBS 1216..1227
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
CDS 1234..2217
/codon_start=1
/gene="luxB"
/product="LuxB luciferase subunit"
/note="from the Photorhabdus luminescens bioluminescence
operon"
/translation="MKFGLFFLNFINSTTVQEQSIVRMQEITEYVDKLNFEQILVYENH
FSDNGVVGAPLTVSGFLLGLTEKIKIGSLNHIITTHHPVRIAEEACLLDQLSEGRFILG
FSDCEKKDEMHFFNRPVEYQQQLFEECYEIINDALTTGYCNPDNDFYSFPKISVNPHAY
TPGGPRKYVTATSHHIVEWAAKKGIPLIFKWDDSNDVRYEYAERYKAVADKYDVDLSEI
DHQLMILVNYNEDSNKAKQETRAFISDYVLEMHPNENFENKLEEIIAENAVGNYTECIT
AAKLAIEKCGAKSVLLSFEPMNDLMSQKNVINIVDDNIKKYHMEYT"
RBS 2229..2240
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
CDS 2247..3689
/codon_start=1
/gene="luxC"
/product="LuxC fatty acid reductase"
/note="from the Photorhabdus luminescens bioluminescence
operon"
/translation="MTKKISFIINGQVEIFPESDDLVQSINFGDNSVYLPILNDSHVKN
IIDCNGNNELRLHNIVNFLYTVGQRWKNEEYSRRRTYIRDLKKYMGYSEEMAKLEANWI
SMILCSKGGLYDVVENELGSRHIMDEWLPQDESYVRAFPKGKSVHLLAGNVPLSGIMSI
LRAILTKNQCIIKTSSTDPFTANALALSFIDVDPNHPITRSLSVIYWPHQGDTSLAKEI
MRHADVIVAWGGPDAINWAVEHAPSYADVIKFGSKKSLCIIDNPVDLTSAATGAAHDVC
FYDQRACFSAQNIYYMGNHYEEFKLALIEKLNLYAHILPNAKKDFDEKAAYSLVQKESL
FAGLKVEVDIHQRWMIIESNAGVEFNQPLGRCVYLHHVDNIEQILPYVQKNKTQTISIF
PWESSFKYRDALALKGAERIVEAGMNNIFRVGGSHDGMRPLQRLVTYISHERPSNYTAK
DVAVEIEQTRFLEEDKFLVFVP"
RBS 3701..3712
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
CDS 3719..4642
/codon_start=1
/gene="luxD"
/product="LuxD acyltransferase"
/note="from the Photorhabdus luminescens bioluminescence
operon"
/translation="MENESKYKTIDHVICVEGNKKIHVWETLPEENSPKRKNAIIIASG
FARRMDHFAGLAEYLSRNGFHVIRYDSLHHVGLSSGTIDEFTMSIGKQSLLAVVDWLTT
RKINNFGMLASSLSARIAYASLSEINASFLITAVGVVNLRYSLERALGFDYLSLPINEL
PDNLDFEGHKLGAEVFARDCLDFGWEDLASTINNMMYLDIPFIAFTANNDNWVKQDEVI
TLLSNIRSNRCKIYSLLGSSHDLSENLVVLRNFYQSVTKAAIAMDNDHLDIDVDITEPS
FEHLTIATVNERRMRIEIENQAISLS"
RBS 4654..4665
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
CDS 4672..5784
/codon_start=1
/gene="luxE"
/product="LuxE"
/note="from the Photorhabdus luminescens bioluminescence
operon"
/translation="MTSYVDKQEITASSEIDDLIFSSDPLVWSYDEQEKIRKKLVLDAF
RNHYKHCREYRHYCQAHKVDDNITEIDDIPVFPTSVFKFTRLLTSQENEIESWFTSSGT
NGLKSQVARDRLSIERLLGSVSYGMKYVGSWFDHQIELVNLGPDRFNAHNIWFKYVMSL
VELLYPTTFTVTEERIDFVKTLNSLERIKNQGKDLCLIGSPYFIYLLCHYMKDKKISFS
GDKSLYIITGGGWKSYEKESLKRDDFNHLLFDTFNLS-DISQIRDIFNQVELNTCFFEDE
MQRKHVPPWVYARALDPETLKPVPDGTPGLMSYMDASATSYPAFIVTDDVGIISREYGK
YPGVLVEILRRVNTRTQKGCALSLTEAFDS"
misc_feature 5788..5808
/label=BioBrick suffix
/note="universal suffix for all parts"
terminator 5859..5945
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 7489..7583
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
CDS complement(7599..8393)
/codon_start=1
/gene="aphA-3"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin"
/translation="MAKMRISPELKKLIEEYRCVKDTEGMSPAKVYKLVGENEILYLKM
TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEGYED
EQSPEKTIELYAECIRLFHSIDISDCPYTNSLDGRLAELDYLLNNDLADVDCENWEEDT
PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
ORIGIN
1 aattcgcggc cgcttctaga ggaaatcata aaaaatttat ttgctttgtg agcggataac
61 aattataata gattcaattg tgagcggata acaattacta gagaaagagg agaaatacta
121 gatgaaattt ggaaactttt tgcttacata ccaacctccc caattttctc aaacagaggt
181 aatgaaacgt ttggttaaat taggtcgcat ctctgaggag tgtggttttg ataccgtatg
241 gttactggag catcatttca cggagtttgg tttgcttggt aacccttatg tcgctgctgc
301 atatttactt ggcgcgacta aaaaattgaa tgtaggaact gccgctattg ttcttcccac
361 agcccatcca gtacgccaac ttgaagatgt gaatttattg gatcaaatgt caaaaggacg
421 atttcggttt ggtatttgcc gagggcttta caacaaggac tttcgcgtat tcggcacaga
481 tatgaataac agtcgcgcct tagcggaatg ctggtacggg ctgataaaga atggcatgac
541 agagggatat atggaagctg ataatgaaca tatcaagttc cataaggtaa aagtaaaccc
601 cgcggcgtat agcagaggtg gcgcaccggt ttatgtggtg gctgaatcag cttcgacgac
661 tgagtgggct gctcaatttg gcctaccgat gatattaagt tggattataa atactaacga
721 aaagaaagca caacttgagc tttataatga agtggctcaa gaatatgggc acgatattca
781 taatatcgac cattgcttat catatataac atctgtagat catgactcaa ttaaagcgaa
841 agagatttgc cggaaatttc tggggcattg gtatgattct tatgtgaatg ctacgactat
901 ttttgatgat tcagaccaaa caagaggtta tgatttcaat aaagggcagt ggcgtgactt
961 tgtattaaaa ggacataaag atactaatcg ccgtattgat tacagttacg aaatcaatcc
1021 cgtgggaacg ccgcaggaat gtattgacat aattcaaaaa gacattgatg ctacaggaat
1081 atcaaatatt tgttgtggat ttgaagctaa tggaacagta gacgaaatta ttgcttccat
1141 gaagctcttc cagtctgatg tcatgccatt tcttaaagaa aaacaacgtt cgctattata
1201 ttaataatac tagagaaaga ggagaaatac tagatgaaat ttggattgtt cttccttaac
1261 ttcatcaatt caacaactgt tcaagaacaa agtatagttc gcatgcagga aataacggag
1321 tatgttgata agttgaattt tgaacagatt ttagtgtatg aaaatcattt ttcagataat
1381 ggtgttgtcg gcgctcctct gactgtttct ggttttctgc tcggtttaac agagaaaatt
1441 aaaattggtt cattaaatca catcattaca actcatcatc ctgtccgcat agcggaggaa
1501 gcttgcttat tggatcagtt aagtgaaggg agatttattt tagggtttag tgattgcgaa
1561 aaaaaagatg aaatgcattt ttttaatcgc ccggttgaat atcaacagca actatttgaa
1621 gagtgttatg aaatcattaa cgatgcttta acaacaggct attgtaatcc agataacgat
1681 ttttatagct tccctaaaat atctgtaaat ccccatgctt atacgccagg cggacctcgg
1741 aaatatgtaa cagcaaccag tcatcatatt gttgagtggg cggccaaaaa aggtattcct
1801 ctcatcttta agtgggatga ttctaatgat gttagatatg aatatgctga aagatataaa
1861 gccgttgcgg ataaatatga cgttgaccta tcagagatag accatcagtt aatgatatta
1921 gttaactata acgaagatag taataaagct aaacaagaga cgcgtgcatt tattagtgat
1981 tatgttcttg aaatgcaccc taatgaaaat ttcgaaaata aacttgaaga aataattgca
2041 gaaaacgctg tcggaaatta tacggagtgt ataactgcgg ctaagttggc aattgaaaag
2101 tgtggtgcga aaagtgtatt gctgtccttt gaaccaatga atgatttgat gagccaaaaa
2161 aatgtaatca atattgttga tgataatatt aagaagtacc acatggaata tacctaataa
2221 tactagagaa agaggagaaa tactagatga ctaaaaaaat ttcattcatt attaacggcc
2281 aggttgaaat ctttcccgaa agtgatgatt tagtgcaatc cattaatttt ggtgataata
2341 gtgtttacct gccaatattg aatgactctc atgtaaaaaa cattattgat tgtaatggaa
2401 ataacgaatt acggttgcat aacattgtca attttctcta tacggtaggg caaagatgga
2461 aaaatgaaga atactcaaga cgcaggacat acattcgtga cttaaaaaaa tatatgggat
2521 attcagaaga aatggctaag ctagaggcca attggatatc tatgatttta tgttctaaag
2581 gcggccttta tgatgttgta gaaaatgaac ttggttctcg ccatatcatg gatgaatggc
2641 tacctcagga tgaaagttat gttcgggctt ttccgaaagg taaatctgta catctgttgg
2701 caggtaatgt tccattatct gggatcatgt ctatattacg cgcaatttta actaagaatc
2761 agtgtattat aaaaacatcg tcaaccgatc cttttaccgc taatgcatta gcgttaagtt
2821 ttattgatgt agaccctaat catccgataa cgcgctcttt atctgttata tattggcccc
2881 accaaggtga tacatcactc gcaaaagaaa ttatgcgaca tgcggatgtt attgtcgctt
2941 ggggagggcc agatgcgatt aattgggcgg tagagcatgc gccatcttat gctgatgtga
3001 ttaaatttgg ttctaaaaag agtctttgca ttatcgataa tcctgttgat ttgacgtccg
3061 cagcgacagg tgcggctcat gatgtttgtt tttacgatca gcgagcttgt ttttctgccc
3121 aaaacatata ttacatggga aatcattatg aggaatttaa gttagcgttg atagaaaaac
3181 ttaatctata tgcgcatata ttaccgaatg ccaaaaaaga ttttgatgaa aaggcggcct
3241 attctttagt tcaaaaagaa agcttgtttg ctggattaaa agtagaggtg gatattcatc
3301 aacgttggat gattattgag tcaaatgcag gtgtggaatt taatcaacca cttggcagat
3361 gtgtgtacct tcatcacgtc gataatattg agcaaatatt gccttatgtt caaaaaaata
3421 agacgcaaac catatctatt tttccttggg agtcatcatt taaatatcga gatgcgttag
3481 cattaaaagg tgcggaaagg attgtagaag caggaatgaa taacatattt cgagttggtg
3541 gatctcatga cggaatgaga ccgttgcaac gattagtgac atatatttct catgaaaggc
3601 catctaacta tacggctaag gatgttgcgg ttgaaataga acagactcga ttcctggaag
3661 aagataagtt ccttgtattt gtcccataat aatactagag aaagaggaga aatactagat
3721 ggaaaatgaa tcaaaatata aaaccatcga ccacgttatt tgtgttgaag gaaataaaaa
3781 aattcatgtt tgggaaacgc tgccagaaga aaacagccca aagagaaaga atgccattat
3841 tattgcgtct ggttttgccc gcaggatgga tcattttgct ggtctggcgg aatatttatc
3901 gcggaatgga tttcatgtga tccgctatga ttcgcttcac cacgttggat tgagttcagg
3961 gacaattgat gaatttacaa tgtctatagg aaagcagagc ttgttagcag tggttgattg
4021 gttaactaca cgaaaaataa ataacttcgg tatgttggct tcaagcttat ctgcgcggat
4081 agcttatgca agcctatctg aaatcaatgc ttcgttttta atcaccgcag tcggtgttgt
4141 taacttaaga tattctcttg aaagagcttt agggtttgat tatctcagtc tacccattaa
4201 tgaattgccg gataatctgg attttgaagg ccataaattg ggtgctgaag tctttgcgag
4261 agattgtctt gattttggtt gggaagattt agcttctaca attaataaca tgatgtatct
4321 tgatataccg tttattgctt ttactgcaaa taacgataat tgggtcaagc aagatgaagt
4381 tatcacattg ttatcaaata ttcgtagtaa tcgatgcaag atatattctt tgttaggaag
4441 ttcgcatgac ttgagtgaaa atttagtggt cctgcgcaat ttttatcaat cggttacgaa
4501 agccgctatc gcgatggata atgatcatct ggatattgat gttgatatta ctgaaccgtc
4561 atttgaacat ttaactattg cgacagtcaa tgaacgccga atgagaattg agattgaaaa
4621 tcaagcaatt tctctgtctt aataatacta gagaaagagg agaaatacta gatgacttca
4681 tatgttgata aacaagaaat tacagcaagc tcagaaattg atgatttgat tttttcgagc
4741 gatccattag tgtggtctta cgacgagcag gaaaaaatca gaaagaaact tgtgcttgat
4801 gcatttcgta atcattataa acattgtcga gaatatcgtc actactgtca ggcacacaaa
4861 gtagatgaca atattacgga aattgatgac atacctgtat tcccaacatc ggtttttaag
4921 tttactcgct tattaacttc tcaggaaaac gagattgaaa gttggtttac cagtagcggc
4981 acgaatggtt taaaaagtca ggtggcgcgt gacagattaa gtattgagag actcttaggc
5041 tctgtgagtt atggcatgaa atatgttggt agttggtttg atcatcaaat agaattagtc
5101 aatttgggac cagatagatt taatgctcat aatatttggt ttaaatatgt tatgagtttg
5161 gtggaattgt tatatcctac gacatttacc gtaacagaag aacgaataga ttttgttaaa
5221 acattgaata gtcttgaacg aataaaaaat caagggaaag atctttgtct tattggttcg
5281 ccatacttta tttatttact ctgccattat atgaaagata aaaaaatctc attttctgga
5341 gataaaagcc tttatatcat aaccggaggc ggctggaaaa gttacgaaaa agaatctctg
5401 aaacgtgatg atttcaatca tcttttattt gatactttca atctcagtga tattagtcag
5461 atccgagata tatttaatca agttgaactc aacacttgtt tctttgagga tgaaatgcag
5521 cgtaaacatg ttccgccgtg ggtatatgcg cgagcgcttg atcctgaaac gttgaaacct
5581 gtacctgatg gaacgccggg gttgatgagt tatatggatg cgtcagcaac cagttatcca
5641 gcatttattg ttaccgatga tgtcgggata attagcagag aatatggtaa gtatcccggc
5701 gtgctcgttg aaattttacg tcgcgtcaat acgaggacgc agaaagggtg tgctttaagc
5761 ttaaccgaag cgtttgatag ttaataatac tagtagcggc cgctgcaggc ctcagggccc
5821 gatcgatgcc gccgcttaat taattaatcc agaggcatca aataaaacga aaggctcagt
5881 cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc ctgagtagga
5941 caaatccgcc gccctagacc tagtgtcatt ttatttcccc cgtttcagca tcaagaacct
6001 ttgcataact tgctctatat ccacactgat aattgccctc aaaccataat ctaaaggcgc
6061 tagagtttgt tgaaacaata tcttttacat cattcgtatt taaaattcca aactccgctc
6121 ccctaaggcg aataaaagcc attaaatctt ttgtatttac caaattatag tcatccacta
6181 tatctaagag taaattcttc aattctcttt tttggctttc atcaagtgtt atatagcggt
6241 caatatcaaa atcattaatg ttcaaaatat cttttttgtc gtatatatgt ttattcttag
6301 caatagcgtc ctttgattca tgagtcaaat attcatatga acctttgata taatcaagta
6361 tctcaacatg agcaactgaa ctattcccca attttcgctt aatcttgttc ctaacgcttt
6421 ctattgttac aggatttcgt gcaatatata taacgtgata gtgtggtttt ttatagtgct
6481 ttccatttcg tataacatca ctactattcc atgtatcttt atcttttttt tcgtccatat
6541 cgtgtaaagg actgacagcc atagatacgc ccaaactctc taatttttcc ttccaatcat
6601 taggaattga gtcaggatat aataaaaatc caaaatttct agctttagta tttttaatag
6661 ccatgatata attaccttat caaaaacaag tagcgaaaac tcgtatcctt ctaaaaacgc
6721 gagctttcgc ttattttttt tgttctgatt cctttcttgc atattcttct atagctaacg
6781 ccgcaaccgc agattttgaa aaaccttttt gtttcgccat atctgttaat tttttatctt
6841 gctcttttgt cagagaaatc ataactcttt ttttcgattc tgaaatcacc atttaaaaaa
6901 ctccaatcaa ataattttat aaagttagtg tatcactttg taatcataaa aacaacaata
6961 aagctactta aatatagatt tataaaaaac gttggcgaaa acgttggcga ttcgttggcg
7021 attgaaaaac cccttaaacc cttgagccag ttgggataga gcgtttttgg cacaaaaatt
7081 ggcactcggc acttaatggg gggtcgtagt acggaagcaa aattcgcttc ctttcccccc
7141 atttttttcc aaattccaaa tttttttcaa aaattttcca gcgctaccgc tcggcaaaat
7201 tgcaagcaat ttttaaaatc aaacccatga gggaatttca ttccctcata ctcccttgag
7261 cctcctccaa ccgaaataga agggcgctgc gcttattatt tcattcagtc atcggctttc
7321 ataatctaac agacaacatc ttcgctgcaa agccacgcta cgctcaaggg cttttacgct
7381 acgataacgc ctgttttaac gattatgccg ataactaaac gaaataaacg ctaaaacgtc
7441 tcagaaacga ttttgagacg ttttaataaa aaatcgccta gtgcttggat tctcaccaat
7501 aaaaaacgcc cggcggcaac cgagcgttct gaacaaatcc agatggagtt ctgaggtcat
7561 tactggatct atcaacagga gtccaagcga gctcggtact aaaacaattc atccagtaaa
7621 atataatatt ttattttctc ccaatcaggc ttgatcccca gtaagtcaaa aaatagctcg
7681 acatactgtt cttccccgat atcctccctg atcgaccgga cgcagaaggc aatgtcatac
7741 cacttgtccg ccctgccgct tctcccaaga tcaataaagc cacttacttt gccatctttc
7801 acaaagatgt tgctgtctcc caggtcgccg tgggaaaaga caagttcctc ttcgggcttt
7861 tccgtcttta aaaaatcata cagctcgcgc ggatctttaa atggagtgtc ttcttcccag
7921 ttttcgcaat ccacatcggc cagatcgtta ttcagtaagt aatccaattc ggctaagcgg
7981 ccgtctaagc tattcgtata gggacaatcc gatatgtcga tggagtgaaa gagcctgatg
8041 cactccgcat acagctcgat agtcttttca gggctttgtt catcttcata cccttccgag
8101 caaaggacgc catcggcctc actcatgagc agattgctcc agccatcatg ccgttcaaag
8161 tgcaggacct ttggaacagg cagctttcct tccagccata gcatcatgtc cttttcccgt
8221 tccacatcat aggtggtccc tttataccgg ctgtccgtca tttttaaata taggatttca
8281 ttttctccca ccagcttata taccttagca ggagacattc cttccgtatc ttttacgcag
8341 cggtattctt cgatcagttt tttcaattcc ggtgatattc tcattttagc catttattat
8401 ttccttcctc ttttctacag tatttaaaga taccccaaga agctaattat aacaagacga
8461 actccaattc actgttcctt gcattctaaa accttaaata cagaaaacag ccttttcaaa
8521 gttgttttca aagttggcgt ataacatagt atcgacggag ccgattttga aaccacaatt
8581 atgatagaat ttgacgtcg
//
质粒菌株产品操作说明书
一、扩增流程
收到产品后，请先根据产品管壁标签来判断产品形式，并在扩增前准确查找该质粒菌株的抗性、感受态和培养温度。
1、质粒干粉（常温运输，存于-20度，90天保质期，请务必转化挑单克隆培养，不要直接使用和测序）
①收到质粒干粉后请先5000rpm离心1min，再加入20μl ddH2O去离子水溶解质粒；
②取1支100μl 感受态于冰上解冻10min，加入2μl质粒，再冰浴30min后，42℃热激60s，不要搅动，再冰浴2min；（从第二步开始均要在超净工作台中无菌操作）
③加入900μl无抗的LB液体培养基，180rpm震荡37℃培养45min；
④6000rpm离心5min，仅留100μl上清液重悬细菌沉淀，并涂布至目标质粒抗性的LB平板上；（可使用本平台的平板涂布专用玻璃珠进行涂布，可以比传统涂布方法获得更多转化子）
⑤将平板正向培养1h，再倒置37℃培养14h。如果要求是30度则培养20h；
（菌落过多则将质粒稀释后再转化。没有菌落则加入10μl质粒转化。另不要直接转表达感受态，要先转克隆感受态，重提质粒后再导入表达感受态）
⑥挑取单菌落至LB液体培养基中，加入对应抗生素，220rpm震荡培养14h，根据实验需要和质粒提取试剂盒说明书提取质粒。
2、甘油菌种（冰袋运输，存于-80℃，保质期90天，请务必划线挑单克隆培养）
四区划线后挑单菌落培养，酵母菌需要先液体复苏再四区划线，再挑单菌落液体培养。
3、穿刺菌种（冰袋运输，存于4℃，保质期7天）
穿刺接种，液体培养后四区划线，再挑单菌落液体培养。
4、菌落平板（冰袋运输，存于4℃，保质期7天）
直接挑取单菌落至液体培养基中。
5、液体质粒（冰袋运输，存于-20℃，保质期90天）
单独提取的液体质粒收到后可直接使用。
6、滤纸质粒（常温运输，存于-20度，90天保质期，请务必转化挑单克隆培养，不要直接使用和测序）
收到货后将滤纸画圈部分剪下放入EP管中，加100ul无菌水将滤纸浸湿并浸泡5min，吸取5ul质粒转化，离心全涂。
二、转化图片
名称：pBAV1k-T5-LUX基因质粒
别称: Plasmid#55800;LuxCDABE
启动子：T5
复制子: pUC
终止子: rnB T1 terminator
质粒分类: 基因文库质粒；其它基因质粒；其它种源质粒
质粒大小：8599bp
原核抗性: Kan
筛选标记：G418
克隆菌株: DH5a
培养条件: 30度
备注: 低拷贝
质粒属性
载体宿主：大肠杆菌
载体用途：蛋白表达
基因种属：
基因类型：ORF
原核抗性：Kan
真核抗性：
荧光蛋白：
质粒简介
Expresses the Photorhabdus luminescens lux operon in bacterial cells.The copy number of pBAV1k plasmids are not well controlled, so purification yields vary. A lacI+ E. coli strain will prevent constitutive expression of the lux operon.
质粒图谱
质粒序列
LOCUS Exported 8599 bp ds-DNA circular SYN 15-AUG-2017
DEFINITION synthetic circular DNA
FEATURES Location/Qualifiers
source 1..8599
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 26..70
/label=T5 promoter
/note="bacteriophage T5 promoter for E. coli RNA
polymerase, with embedded lac operator"
protein_bind 46..62
/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)."
protein_bind 78..94
/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 104..115
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
CDS 122..1204
/codon_start=1
/gene="luxA"
/product="LuxA luciferase subunit"
/note="from the Photorhabdus luminescens bioluminescence
operon"
/translation="MKFGNFLLTYQPPQFSQTEVMKRLVKLGRISEECGFDTVWLLEHH
FTEFGLLGNPYVAAAYLLGATKKLNVGTAAIVLPTAHPVRQLEDVNLLDQMSKGRFRFG
ICRGLYNKDFRVFGTDMNNSRALAECWYGLIKNGMTEGYMEADNEHIKFHKVKVNPAAY
SRGGAPVYVVAESASTTEWAAQFGLPMILSWIINTNEKKAQLELYNEVAQEYGHDIHNI
DHCLSYITSVDHDSIKAKEICRKFLGHWYDSYVNATTIFDDSDQTRGYDFNKGQWRDFV
LKGHKDTNRRIDYSYEINPVGTPQECIDIIQKDIDATGISNICCGFEANGTVDEIIASM
KLFQSDVMPFLKEKQRSLLY"
RBS 1216..1227
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
CDS 1234..2217
/codon_start=1
/gene="luxB"
/product="LuxB luciferase subunit"
/note="from the Photorhabdus luminescens bioluminescence
operon"
/translation="MKFGLFFLNFINSTTVQEQSIVRMQEITEYVDKLNFEQILVYENH
FSDNGVVGAPLTVSGFLLGLTEKIKIGSLNHIITTHHPVRIAEEACLLDQLSEGRFILG
FSDCEKKDEMHFFNRPVEYQQQLFEECYEIINDALTTGYCNPDNDFYSFPKISVNPHAY
TPGGPRKYVTATSHHIVEWAAKKGIPLIFKWDDSNDVRYEYAERYKAVADKYDVDLSEI
DHQLMILVNYNEDSNKAKQETRAFISDYVLEMHPNENFENKLEEIIAENAVGNYTECIT
AAKLAIEKCGAKSVLLSFEPMNDLMSQKNVINIVDDNIKKYHMEYT"
RBS 2229..2240
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
CDS 2247..3689
/codon_start=1
/gene="luxC"
/product="LuxC fatty acid reductase"
/note="from the Photorhabdus luminescens bioluminescence
operon"
/translation="MTKKISFIINGQVEIFPESDDLVQSINFGDNSVYLPILNDSHVKN
IIDCNGNNELRLHNIVNFLYTVGQRWKNEEYSRRRTYIRDLKKYMGYSEEMAKLEANWI
SMILCSKGGLYDVVENELGSRHIMDEWLPQDESYVRAFPKGKSVHLLAGNVPLSGIMSI
LRAILTKNQCIIKTSSTDPFTANALALSFIDVDPNHPITRSLSVIYWPHQGDTSLAKEI
MRHADVIVAWGGPDAINWAVEHAPSYADVIKFGSKKSLCIIDNPVDLTSAATGAAHDVC
FYDQRACFSAQNIYYMGNHYEEFKLALIEKLNLYAHILPNAKKDFDEKAAYSLVQKESL
FAGLKVEVDIHQRWMIIESNAGVEFNQPLGRCVYLHHVDNIEQILPYVQKNKTQTISIF
PWESSFKYRDALALKGAERIVEAGMNNIFRVGGSHDGMRPLQRLVTYISHERPSNYTAK
DVAVEIEQTRFLEEDKFLVFVP"
RBS 3701..3712
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
CDS 3719..4642
/codon_start=1
/gene="luxD"
/product="LuxD acyltransferase"
/note="from the Photorhabdus luminescens bioluminescence
operon"
/translation="MENESKYKTIDHVICVEGNKKIHVWETLPEENSPKRKNAIIIASG
FARRMDHFAGLAEYLSRNGFHVIRYDSLHHVGLSSGTIDEFTMSIGKQSLLAVVDWLTT
RKINNFGMLASSLSARIAYASLSEINASFLITAVGVVNLRYSLERALGFDYLSLPINEL
PDNLDFEGHKLGAEVFARDCLDFGWEDLASTINNMMYLDIPFIAFTANNDNWVKQDEVI
TLLSNIRSNRCKIYSLLGSSHDLSENLVVLRNFYQSVTKAAIAMDNDHLDIDVDITEPS
FEHLTIATVNERRMRIEIENQAISLS"
RBS 4654..4665
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
CDS 4672..5784
/codon_start=1
/gene="luxE"
/product="LuxE"
/note="from the Photorhabdus luminescens bioluminescence
operon"
/translation="MTSYVDKQEITASSEIDDLIFSSDPLVWSYDEQEKIRKKLVLDAF
RNHYKHCREYRHYCQAHKVDDNITEIDDIPVFPTSVFKFTRLLTSQENEIESWFTSSGT
NGLKSQVARDRLSIERLLGSVSYGMKYVGSWFDHQIELVNLGPDRFNAHNIWFKYVMSL
VELLYPTTFTVTEERIDFVKTLNSLERIKNQGKDLCLIGSPYFIYLLCHYMKDKKISFS
GDKSLYIITGGGWKSYEKESLKRDDFNHLLFDTFNLS-DISQIRDIFNQVELNTCFFEDE
MQRKHVPPWVYARALDPETLKPVPDGTPGLMSYMDASATSYPAFIVTDDVGIISREYGK
YPGVLVEILRRVNTRTQKGCALSLTEAFDS"
misc_feature 5788..5808
/label=BioBrick suffix
/note="universal suffix for all parts"
terminator 5859..5945
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 7489..7583
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
CDS complement(7599..8393)
/codon_start=1
/gene="aphA-3"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin"
/translation="MAKMRISPELKKLIEEYRCVKDTEGMSPAKVYKLVGENEILYLKM
TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEGYED
EQSPEKTIELYAECIRLFHSIDISDCPYTNSLDGRLAELDYLLNNDLADVDCENWEEDT
PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
ORIGIN
1 aattcgcggc cgcttctaga ggaaatcata aaaaatttat ttgctttgtg agcggataac
61 aattataata gattcaattg tgagcggata acaattacta gagaaagagg agaaatacta
121 gatgaaattt ggaaactttt tgcttacata ccaacctccc caattttctc aaacagaggt
181 aatgaaacgt ttggttaaat taggtcgcat ctctgaggag tgtggttttg ataccgtatg
241 gttactggag catcatttca cggagtttgg tttgcttggt aacccttatg tcgctgctgc
301 atatttactt ggcgcgacta aaaaattgaa tgtaggaact gccgctattg ttcttcccac
361 agcccatcca gtacgccaac ttgaagatgt gaatttattg gatcaaatgt caaaaggacg
421 atttcggttt ggtatttgcc gagggcttta caacaaggac tttcgcgtat tcggcacaga
481 tatgaataac agtcgcgcct tagcggaatg ctggtacggg ctgataaaga atggcatgac
541 agagggatat atggaagctg ataatgaaca tatcaagttc cataaggtaa aagtaaaccc
601 cgcggcgtat agcagaggtg gcgcaccggt ttatgtggtg gctgaatcag cttcgacgac
661 tgagtgggct gctcaatttg gcctaccgat gatattaagt tggattataa atactaacga
721 aaagaaagca caacttgagc tttataatga agtggctcaa gaatatgggc acgatattca
781 taatatcgac cattgcttat catatataac atctgtagat catgactcaa ttaaagcgaa
841 agagatttgc cggaaatttc tggggcattg gtatgattct tatgtgaatg ctacgactat
901 ttttgatgat tcagaccaaa caagaggtta tgatttcaat aaagggcagt ggcgtgactt
961 tgtattaaaa ggacataaag atactaatcg ccgtattgat tacagttacg aaatcaatcc
1021 cgtgggaacg ccgcaggaat gtattgacat aattcaaaaa gacattgatg ctacaggaat
1081 atcaaatatt tgttgtggat ttgaagctaa tggaacagta gacgaaatta ttgcttccat
1141 gaagctcttc cagtctgatg tcatgccatt tcttaaagaa aaacaacgtt cgctattata
1201 ttaataatac tagagaaaga ggagaaatac tagatgaaat ttggattgtt cttccttaac
1261 ttcatcaatt caacaactgt tcaagaacaa agtatagttc gcatgcagga aataacggag
1321 tatgttgata agttgaattt tgaacagatt ttagtgtatg aaaatcattt ttcagataat
1381 ggtgttgtcg gcgctcctct gactgtttct ggttttctgc tcggtttaac agagaaaatt
1441 aaaattggtt cattaaatca catcattaca actcatcatc ctgtccgcat agcggaggaa
1501 gcttgcttat tggatcagtt aagtgaaggg agatttattt tagggtttag tgattgcgaa
1561 aaaaaagatg aaatgcattt ttttaatcgc ccggttgaat atcaacagca actatttgaa
1621 gagtgttatg aaatcattaa cgatgcttta acaacaggct attgtaatcc agataacgat
1681 ttttatagct tccctaaaat atctgtaaat ccccatgctt atacgccagg cggacctcgg
1741 aaatatgtaa cagcaaccag tcatcatatt gttgagtggg cggccaaaaa aggtattcct
1801 ctcatcttta agtgggatga ttctaatgat gttagatatg aatatgctga aagatataaa
1861 gccgttgcgg ataaatatga cgttgaccta tcagagatag accatcagtt aatgatatta
1921 gttaactata acgaagatag taataaagct aaacaagaga cgcgtgcatt tattagtgat
1981 tatgttcttg aaatgcaccc taatgaaaat ttcgaaaata aacttgaaga aataattgca
2041 gaaaacgctg tcggaaatta tacggagtgt ataactgcgg ctaagttggc aattgaaaag
2101 tgtggtgcga aaagtgtatt gctgtccttt gaaccaatga atgatttgat gagccaaaaa
2161 aatgtaatca atattgttga tgataatatt aagaagtacc acatggaata tacctaataa
2221 tactagagaa agaggagaaa tactagatga ctaaaaaaat ttcattcatt attaacggcc
2281 aggttgaaat ctttcccgaa agtgatgatt tagtgcaatc cattaatttt ggtgataata
2341 gtgtttacct gccaatattg aatgactctc atgtaaaaaa cattattgat tgtaatggaa
2401 ataacgaatt acggttgcat aacattgtca attttctcta tacggtaggg caaagatgga
2461 aaaatgaaga atactcaaga cgcaggacat acattcgtga cttaaaaaaa tatatgggat
2521 attcagaaga aatggctaag ctagaggcca attggatatc tatgatttta tgttctaaag
2581 gcggccttta tgatgttgta gaaaatgaac ttggttctcg ccatatcatg gatgaatggc
2641 tacctcagga tgaaagttat gttcgggctt ttccgaaagg taaatctgta catctgttgg
2701 caggtaatgt tccattatct gggatcatgt ctatattacg cgcaatttta actaagaatc
2761 agtgtattat aaaaacatcg tcaaccgatc cttttaccgc taatgcatta gcgttaagtt
2821 ttattgatgt agaccctaat catccgataa cgcgctcttt atctgttata tattggcccc
2881 accaaggtga tacatcactc gcaaaagaaa ttatgcgaca tgcggatgtt attgtcgctt
2941 ggggagggcc agatgcgatt aattgggcgg tagagcatgc gccatcttat gctgatgtga
3001 ttaaatttgg ttctaaaaag agtctttgca ttatcgataa tcctgttgat ttgacgtccg
3061 cagcgacagg tgcggctcat gatgtttgtt tttacgatca gcgagcttgt ttttctgccc
3121 aaaacatata ttacatggga aatcattatg aggaatttaa gttagcgttg atagaaaaac
3181 ttaatctata tgcgcatata ttaccgaatg ccaaaaaaga ttttgatgaa aaggcggcct
3241 attctttagt tcaaaaagaa agcttgtttg ctggattaaa agtagaggtg gatattcatc
3301 aacgttggat gattattgag tcaaatgcag gtgtggaatt taatcaacca cttggcagat
3361 gtgtgtacct tcatcacgtc gataatattg agcaaatatt gccttatgtt caaaaaaata
3421 agacgcaaac catatctatt tttccttggg agtcatcatt taaatatcga gatgcgttag
3481 cattaaaagg tgcggaaagg attgtagaag caggaatgaa taacatattt cgagttggtg
3541 gatctcatga cggaatgaga ccgttgcaac gattagtgac atatatttct catgaaaggc
3601 catctaacta tacggctaag gatgttgcgg ttgaaataga acagactcga ttcctggaag
3661 aagataagtt ccttgtattt gtcccataat aatactagag aaagaggaga aatactagat
3721 ggaaaatgaa tcaaaatata aaaccatcga ccacgttatt tgtgttgaag gaaataaaaa
3781 aattcatgtt tgggaaacgc tgccagaaga aaacagccca aagagaaaga atgccattat
3841 tattgcgtct ggttttgccc gcaggatgga tcattttgct ggtctggcgg aatatttatc
3901 gcggaatgga tttcatgtga tccgctatga ttcgcttcac cacgttggat tgagttcagg
3961 gacaattgat gaatttacaa tgtctatagg aaagcagagc ttgttagcag tggttgattg
4021 gttaactaca cgaaaaataa ataacttcgg tatgttggct tcaagcttat ctgcgcggat
4081 agcttatgca agcctatctg aaatcaatgc ttcgttttta atcaccgcag tcggtgttgt
4141 taacttaaga tattctcttg aaagagcttt agggtttgat tatctcagtc tacccattaa
4201 tgaattgccg gataatctgg attttgaagg ccataaattg ggtgctgaag tctttgcgag
4261 agattgtctt gattttggtt gggaagattt agcttctaca attaataaca tgatgtatct
4321 tgatataccg tttattgctt ttactgcaaa taacgataat tgggtcaagc aagatgaagt
4381 tatcacattg ttatcaaata ttcgtagtaa tcgatgcaag atatattctt tgttaggaag
4441 ttcgcatgac ttgagtgaaa atttagtggt cctgcgcaat ttttatcaat cggttacgaa
4501 agccgctatc gcgatggata atgatcatct ggatattgat gttgatatta ctgaaccgtc
4561 atttgaacat ttaactattg cgacagtcaa tgaacgccga atgagaattg agattgaaaa
4621 tcaagcaatt tctctgtctt aataatacta gagaaagagg agaaatacta gatgacttca
4681 tatgttgata aacaagaaat tacagcaagc tcagaaattg atgatttgat tttttcgagc
4741 gatccattag tgtggtctta cgacgagcag gaaaaaatca gaaagaaact tgtgcttgat
4801 gcatttcgta atcattataa acattgtcga gaatatcgtc actactgtca ggcacacaaa
4861 gtagatgaca atattacgga aattgatgac atacctgtat tcccaacatc ggtttttaag
4921 tttactcgct tattaacttc tcaggaaaac gagattgaaa gttggtttac cagtagcggc
4981 acgaatggtt taaaaagtca ggtggcgcgt gacagattaa gtattgagag actcttaggc
5041 tctgtgagtt atggcatgaa atatgttggt agttggtttg atcatcaaat agaattagtc
5101 aatttgggac cagatagatt taatgctcat aatatttggt ttaaatatgt tatgagtttg
5161 gtggaattgt tatatcctac gacatttacc gtaacagaag aacgaataga ttttgttaaa
5221 acattgaata gtcttgaacg aataaaaaat caagggaaag atctttgtct tattggttcg
5281 ccatacttta tttatttact ctgccattat atgaaagata aaaaaatctc attttctgga
5341 gataaaagcc tttatatcat aaccggaggc ggctggaaaa gttacgaaaa agaatctctg
5401 aaacgtgatg atttcaatca tcttttattt gatactttca atctcagtga tattagtcag
5461 atccgagata tatttaatca agttgaactc aacacttgtt tctttgagga tgaaatgcag
5521 cgtaaacatg ttccgccgtg ggtatatgcg cgagcgcttg atcctgaaac gttgaaacct
5581 gtacctgatg gaacgccggg gttgatgagt tatatggatg cgtcagcaac cagttatcca
5641 gcatttattg ttaccgatga tgtcgggata attagcagag aatatggtaa gtatcccggc
5701 gtgctcgttg aaattttacg tcgcgtcaat acgaggacgc agaaagggtg tgctttaagc
5761 ttaaccgaag cgtttgatag ttaataatac tagtagcggc cgctgcaggc ctcagggccc
5821 gatcgatgcc gccgcttaat taattaatcc agaggcatca aataaaacga aaggctcagt
5881 cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc ctgagtagga
5941 caaatccgcc gccctagacc tagtgtcatt ttatttcccc cgtttcagca tcaagaacct
6001 ttgcataact tgctctatat ccacactgat aattgccctc aaaccataat ctaaaggcgc
6061 tagagtttgt tgaaacaata tcttttacat cattcgtatt taaaattcca aactccgctc
6121 ccctaaggcg aataaaagcc attaaatctt ttgtatttac caaattatag tcatccacta
6181 tatctaagag taaattcttc aattctcttt tttggctttc atcaagtgtt atatagcggt
6241 caatatcaaa atcattaatg ttcaaaatat cttttttgtc gtatatatgt ttattcttag
6301 caatagcgtc ctttgattca tgagtcaaat attcatatga acctttgata taatcaagta
6361 tctcaacatg agcaactgaa ctattcccca attttcgctt aatcttgttc ctaacgcttt
6421 ctattgttac aggatttcgt gcaatatata taacgtgata gtgtggtttt ttatagtgct
6481 ttccatttcg tataacatca ctactattcc atgtatcttt atcttttttt tcgtccatat
6541 cgtgtaaagg actgacagcc atagatacgc ccaaactctc taatttttcc ttccaatcat
6601 taggaattga gtcaggatat aataaaaatc caaaatttct agctttagta tttttaatag
6661 ccatgatata attaccttat caaaaacaag tagcgaaaac tcgtatcctt ctaaaaacgc
6721 gagctttcgc ttattttttt tgttctgatt cctttcttgc atattcttct atagctaacg
6781 ccgcaaccgc agattttgaa aaaccttttt gtttcgccat atctgttaat tttttatctt
6841 gctcttttgt cagagaaatc ataactcttt ttttcgattc tgaaatcacc atttaaaaaa
6901 ctccaatcaa ataattttat aaagttagtg tatcactttg taatcataaa aacaacaata
6961 aagctactta aatatagatt tataaaaaac gttggcgaaa acgttggcga ttcgttggcg
7021 attgaaaaac cccttaaacc cttgagccag ttgggataga gcgtttttgg cacaaaaatt
7081 ggcactcggc acttaatggg gggtcgtagt acggaagcaa aattcgcttc ctttcccccc
7141 atttttttcc aaattccaaa tttttttcaa aaattttcca gcgctaccgc tcggcaaaat
7201 tgcaagcaat ttttaaaatc aaacccatga gggaatttca ttccctcata ctcccttgag
7261 cctcctccaa ccgaaataga agggcgctgc gcttattatt tcattcagtc atcggctttc
7321 ataatctaac agacaacatc ttcgctgcaa agccacgcta cgctcaaggg cttttacgct
7381 acgataacgc ctgttttaac gattatgccg ataactaaac gaaataaacg ctaaaacgtc
7441 tcagaaacga ttttgagacg ttttaataaa aaatcgccta gtgcttggat tctcaccaat
7501 aaaaaacgcc cggcggcaac cgagcgttct gaacaaatcc agatggagtt ctgaggtcat
7561 tactggatct atcaacagga gtccaagcga gctcggtact aaaacaattc atccagtaaa
7621 atataatatt ttattttctc ccaatcaggc ttgatcccca gtaagtcaaa aaatagctcg
7681 acatactgtt cttccccgat atcctccctg atcgaccgga cgcagaaggc aatgtcatac
7741 cacttgtccg ccctgccgct tctcccaaga tcaataaagc cacttacttt gccatctttc
7801 acaaagatgt tgctgtctcc caggtcgccg tgggaaaaga caagttcctc ttcgggcttt
7861 tccgtcttta aaaaatcata cagctcgcgc ggatctttaa atggagtgtc ttcttcccag
7921 ttttcgcaat ccacatcggc cagatcgtta ttcagtaagt aatccaattc ggctaagcgg
7981 ccgtctaagc tattcgtata gggacaatcc gatatgtcga tggagtgaaa gagcctgatg
8041 cactccgcat acagctcgat agtcttttca gggctttgtt catcttcata cccttccgag
8101 caaaggacgc catcggcctc actcatgagc agattgctcc agccatcatg ccgttcaaag
8161 tgcaggacct ttggaacagg cagctttcct tccagccata gcatcatgtc cttttcccgt
8221 tccacatcat aggtggtccc tttataccgg ctgtccgtca tttttaaata taggatttca
8281 ttttctccca ccagcttata taccttagca ggagacattc cttccgtatc ttttacgcag
8341 cggtattctt cgatcagttt tttcaattcc ggtgatattc tcattttagc catttattat
8401 ttccttcctc ttttctacag tatttaaaga taccccaaga agctaattat aacaagacga
8461 actccaattc actgttcctt gcattctaaa accttaaata cagaaaacag ccttttcaaa
8521 gttgttttca aagttggcgt ataacatagt atcgacggag ccgattttga aaccacaatt
8581 atgatagaat ttgacgtcg
//
质粒菌株产品操作说明书
一、扩增流程
收到产品后，请先根据产品管壁标签来判断产品形式，并在扩增前准确查找该质粒菌株的抗性、感受态和培养温度。
1、质粒干粉（常温运输，存于-20度，90天保质期，请务必转化挑单克隆培养，不要直接使用和测序）
①收到质粒干粉后请先5000rpm离心1min，再加入20μl ddH2O去离子水溶解质粒；
②取1支100μl 感受态于冰上解冻10min，加入2μl质粒，再冰浴30min后，42℃热激60s，不要搅动，再冰浴2min；（从第二步开始均要在超净工作台中无菌操作）
③加入900μl无抗的LB液体培养基，180rpm震荡37℃培养45min；
④6000rpm离心5min，仅留100μl上清液重悬细菌沉淀，并涂布至目标质粒抗性的LB平板上；（可使用本平台的平板涂布专用玻璃珠进行涂布，可以比传统涂布方法获得更多转化子）
⑤将平板正向培养1h，再倒置37℃培养14h。如果要求是30度则培养20h；
（菌落过多则将质粒稀释后再转化。没有菌落则加入10μl质粒转化。另不要直接转表达感受态，要先转克隆感受态，重提质粒后再导入表达感受态）
⑥挑取单菌落至LB液体培养基中，加入对应抗生素，220rpm震荡培养14h，根据实验需要和质粒提取试剂盒说明书提取质粒。
2、甘油菌种（冰袋运输，存于-80℃，保质期90天，请务必划线挑单克隆培养）
四区划线后挑单菌落培养，酵母菌需要先液体复苏再四区划线，再挑单菌落液体培养。
3、穿刺菌种（冰袋运输，存于4℃，保质期7天）
穿刺接种，液体培养后四区划线，再挑单菌落液体培养。
4、菌落平板（冰袋运输，存于4℃，保质期7天）
直接挑取单菌落至液体培养基中。
5、液体质粒（冰袋运输，存于-20℃，保质期90天）
单独提取的液体质粒收到后可直接使用。
6、滤纸质粒（常温运输，存于-20度，90天保质期，请务必转化挑单克隆培养，不要直接使用和测序）
收到货后将滤纸画圈部分剪下放入EP管中，加100ul无菌水将滤纸浸湿并浸泡5min，吸取5ul质粒转化，离心全涂。
二、转化图片
| P3821/pCMV-SPORT6-FAM184A人源基因质粒 |
| P3822/pCMV-SPORT6-GALNT15（1同义突变） |
| P3823/pCMV-SPORT6-ZC2HC1A人源基因质粒 |
| P0637/pCR4-MME人源基因质粒 |
| P0645/pCR4-IL12A人源基因质粒 |
| P0646/pCR4-PKD2人源基因质粒 |
| P4957/pCR-XL-TOPO-EFCAB4B人源基因质粒 |
| P0647/pDNR-Dual-IL12B人源基因质粒 |
| P4822/pDNR-LIB-PABPN1（2点突变）人源基因质粒 |
风险提示：丁香通仅作为第三方平台，为商家信息发布提供平台空间。用户咨询产品时请注意保护个人信息及财产安全，合理判断，谨慎选购商品，商家和用户对交易行为负责。对于医疗器械类产品，请先查证核实企业经营资质和医疗器械产品注册证情况。
- 作者
- 内容
- 询问日期
文献和实验相关实验
），绿色荧光蛋白（GFP），荧光素酶（Lux或Luc）以及b-半乳糖苷酶（b-gal）。可以使用简单的非同位素方法检测b-gal的表达以测定转染效率和活性。pCMV SPORT- bgal质粒包含CMV启动子调控下的LacZ基因，转染入真核细胞内后可以直接表达bgal。结合简单的检测步骤，可以做为监测转染条件的一种方便灵敏的方法。 09稳定转染细胞系的筛选 用载体中所含的选择标志进行筛选是建立稳定转染细胞系最常用的方法。抗生素抗性基因可以与目的基因在同一个质粒上，也可以在不同的质粒上。如果两个不同
表达不同于其它一些实验，比如：提取质粒、PCR、电镜切片，这些人为控制的因素比较多，出问题相对来说也比较好分析。表达呢，你把质粒克隆好啦，交给细胞，然后有些事情就不全是你要怎样就怎样了。原核表达在表达当中来说还是比较简单，细菌培养条件简单、生长速度快，需要的仪器和培养基都比较便宜。当然，它也存在一些缺乏高级修饰、细胞内部还原性过高等缺点。原核表达从一开始的设计就非常重要，所谓好的开始是成功的一半。做足准备功夫，可是省去很多将来后悔的事情。首先，我们要根据是否要求可溶将载体分成两大类，如果希望
2）。 图2 使用BLOCK-iT™ Complete Dicer RNAi Kit得到＞80％的靶定基因阻断 经Dicer酶切的，来源于β-半乳糖苷酶或者荧光素酶的dsRNA产生siRNA（d-siRNA）库，用来确定d-siRNA特异阻断基因活性的效果。GripTite™ 293 MSR 细胞共转染pcDNA™1.2/V5-GW/lacZ阳性对照质粒和pcDNA™5/FRT/luc，分别独自表达β-gal或者荧光素酶报告子，或者50ng luc 或者lacZ d-siRNA。对于每一种报告子
pBAV1k-T5-LUX基因质粒
￥100 - 1000
