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pRed-FLp

产品货号 P0789
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基本信息

产品简介

Instructions Ⅰ. Plasmid dry powder (stored at minus 20 degrees, shelf life 90 days, transportation at room temperature) 1. After receiving the dry powder plasmid, please centrifuge at 5000rpm for 1min, and then add 20μl of sterile water to dissolve the plasmid; 2. Thaw the corresponding competent on an ice box, take 2 μl of plasmid and add it to 100 μl of competent for 30min; 3. 42 degrees heat shock for 60s, ice bath for 2min, add 900μl of antibiotic-free LB medium, and culture at 180rpm shaking for 45min; 4. Centrifuge at 6000rpm for 3min, leave 100μl of supernatant to resuspend, and then coat it on the corresponding resistant LB plate; 5.After inverting overnight, if there are too many colonies, the plasmid will be diluted for transformation, and if there are no colonies, 10ul plasmid will be taken for transformation; 6. Pick single colonies into the corresponding resistant LB liquid medium overnight shaking culture, and extract the plasmid as needed. Ⅱ. Liquid plasmid (minus 20 degrees storage, shelf life 90 days, ice pack transportation). 1. After receiving the liquid plasmid, please shake the preservation tube so that the liquid plasmid does not adsorb on the tube wall; 2.Take an appropriate amount of plasmid according to the experimental needs, and store the rest in a minus 20 degree freezer. Ⅲ. Glycerol strains (stored at minus 80 degrees, shelf life of 90 days, ice pack transportation). 1. After receiving the glycerol strain, please shake the preservation tube so that the bacterial solution does not adsorb on the tube wall; 2. Use the inoculation loop to mark the line in four areas on the solid plate, and add the corresponding resistance if there is resistance; 3. Inverted culture in constant temperature incubator for 12~16h; 4. Be sure to pick a single colony and inoculate it in the liquid medium, and add the corresponding resistance if there is resistance; 5. Shake the shaker and culture overnight, and culture the strains according to the needs of the experiment.

产品声明

  1. For Research Use Only
    All products available through the MiaoLing Plasmid Platform are intended for research use only. They are not intended for clinical trials or for use in humans, including oral administration, injection, or therapeutic use in vivo or ex vivo.
  2. Plasmid Sequence and Experimental Performance
    Plasmid products are provided and sold based on the corresponding reference/template plasmid. MiaoLing guarantees the accuracy of the specified gene sequence based on our sequencing results, but does not guarantee downstream experimental performance, including gene expression, transfection efficiency, protein expression, or other experimental outcomes.
  3. Sequence Verification of Certain Vector Products
    For certain vector products, even the original manufacturer may not have fully verified the complete plasmid sequence. If the sequence of the key functional regions has been confirmed to be correct, the product will be considered a correct and usable vector for research purposes.
  4. After-Sales Period for Small-Pack Plasmid Products
    The after-sales service period for small-pack plasmid products is three months from the date of receipt. Customers are advised to transform and process the plasmid promptly after receipt. After the three-month period, a new purchase may be required for further support or replacement.
  5. Research-Related Risks and Limitation of Liability
    Scientific research is inherently exploratory and involves a significant degree of uncertainty. To the maximum extent permitted by applicable law, MiaoLing shall not be liable for any additional economic losses, indirect losses, or other liabilities arising from the use of our products or from research outcomes.
  6. Citation in Chinese Publications
    For publications in Chinese, please cite the product as follows:
    产品名称（产品货号）由淼灵质粒平台提供。
  7. Citation in English Publications
    For publications in English, please cite the product as follows:
    “Product Name (Product Item Number) was obtained from MiaoLing Plasmid, China.”

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产品文件

P0789_Sequencing report.zip

质粒元件

f1 ori 23..478 /direction=RIGHT /note="f1噬菌体复制起点；箭头表示（+）链合成的方向" AmpR promoter 504..608 /gene="bla" AmpR 609..1469 /codon_start=1 /gene="bla" /product="beta-lactamase" /note="对氨苄青霉素、羧苄青霉素和相关抗生素产生耐药性" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGECGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" TetR 1477..2100 /codon_start=1 /gene="tetR" /product="tetracycline repressor TetR" /note="TetR与四环素算子tetO结合以抑制转录。这种抑制可以通过添加四环素或强力霉素来缓解。" /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS" CDS complement(2143..3093) /codon_start=1 /gene="rep101" /product="RepA protein needed for replication with the pSC101 origin" /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE RTVSFTYNQYVQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI" pSC101 ori 3141..3363 /note="需要Rep101蛋白的低拷贝复制起点" Feature 30 3922..3923 tetR/tetA promoters 3929..3984 /gene="tetR" /note="细菌tetR和tetA的重叠启动子" tet operator 3935..3953 /bound_moiety="tetracycline repressor TetR" /note="tetR和tetA基因的细菌操作员O1" tet operator 3965..3983 /gene="tetO" /bound_moiety="tetracycline repressor TetR" /note="tetR和tetA基因的细菌操作员O2" FLP 4032..5303 /codon_start=1 /product="site-specific recombinase" /note="FLP是一种来自酿酒酵母的位点特异性重组酶。重组发生在FRT序列上。" /translation="MPQFDILCKTPPKVLVRQFVERFERPSGEKIALCAAELTYLCWMI THNGTAIKRATFMSYNTIISNSLSFDIVNKSLQFKYKTQKATILEASLKKLIPAWEFTI IPYYGQKHQSDITDIVSSLQLQFESSEEADKGNSHSKKMLKALLSEGESIWEITEKILN SFEYTSRFTKTKTLYQFLFLATFINCGRFSDIKNVDPKSFKLVQNKYLGVIIQCLVTET KTSVSRHIYFFSARGRIDPLVYLDEFLRNSEPVLKRVNRTGNSSSNKQEYQLLKDNLVR SYNKALKKNAPYSIFAIKNGPKSHIGRHLMTSFLSMKGLTELTNVVGNWSDKRASAVAR TTYTHQITAIPDHYFALVSRYYAYDPISKEMIALKDETNPIEEWQHIEQLKGSAEGSIR YPAWNGIISQEVLDYLSSYINRRI" TpR complement(5465..5701) /codon_start=1 /gene="dhfrIIb from plasmid R388" /product="E. coli plasmid-associated dihydrofolate reductase" /note="赋予对甲氧苄啶的抗性" /translation="MGQSSDEANAPVAGQFALPLSATFGLGDRVRKKSGAAWQGQVVGW YCTKLTPEGYAVESESHPGSVQIYPVAALERVA" Pc promoter complement(6028..6056) /gene="intI1 (promoter lies within the coding sequence)" /note="1类整合子启动子" rhaR complement(6087..6935) /codon_start=1 /gene="E. coli rhaR" /product="transcriptional activator of rhaR and rhaS" /note="鼠李糖依赖性激活鼠李糖B启动子所需" /translation="MAHQLKLLKDDFFASDQQAVAVADRYPQDVFAEHTHDFCELVIVW RGNGLHVLNDRPYRITRGDLFYIHADDKHSYASVNDLVLQNIIYCPERLKLNLDWQGAI PGFNASAGQPHWRLGSMGMAQARQVIGQLEHESSQHVPFANEMAELLFGQLVMLLNRHR YTSDSLPPTSSETLLDKLITRLAASLKSPFALDKFCDEASCSERVLRQQFRQQTGMTIN QYLRQVRVCHAQYLLQHSRLLISDISTECGFEDSNYFSVVFTRETGMTPSQWRHLNSQK D" rhaS complement(7009..7845) /codon_start=1 /gene="E. coli rhaS" /product="positive regulator of the rhaB promoter " /note="L-鼠李糖激活" /translation="MTVLHSVDFFPSGNASVAIEPRLPQADFPEHHHDFHEIVIVEHGT GIHVFNGQPYTITGGTVCFVRDHDRHLYEHTDNLCLTNVLYRSPDRFQFLAGLNQLLPQ ELDGQYPSHWRVNHSVLQQVRQLVAQMEQQEGENDLPSTASREILFMQLLLLLRKSSLQ ENLENSASRLNLLLAWLEDHFADEVNWDAVADQFSLSLRTLHRQLKQQTGLTPQRYLNR LRLMKARHLLRHSEASVTDIAYRCGFSDSNHFSTLFRREFNWSPRDIRQGRDGFLQ" rhaB promoter 7991..8109 /note="大肠杆菌rhaBAD操纵子的启动子，在RhaR和RhaS存在的情况下，与L-鼠李糖紧密诱导，与D-葡萄糖抑制（Giacalone等人，2006）" CDS 8133..8549 /codon_start=1 /gene="gam" /product="inhibitor of the host RecBCD nuclease in the lambda Red system" /note="保护双链DNA底物进行重组" /translation="MDINTETEIKQKHSLTPFPVFLISPAFRGRYFHSYFRSSAMNAYY IQDRLEAQSWARHYQQLAREEKEAELADDMEKGLPQHLFESLCIDHLQRHGASKKSITR AFDDDVEFQERMAEHIRYMVETIAHHQVDIDSEV" CDS 8555..9340 /codon_start=1 /gene="bet" /product="single-stranded DNA binding recombinase in the lambda Red system" /note="介导单链DNA与互补靶标的配对" /translation="MSTALATLAGKLAERVGMDSVDPQELITTLRQTAFKGDASDAQFI ALLIVANQYGLNPWTKEIYAFPDKQNGIVPVVGVDGWSRIINENQQFDGMDFEQDNESC TCRIYRKDRNHPICVTEWMDECRREPFKTREGREITGPWQSHPKRMLRHKAMIQCARLA FGFAGIYDKDEAERIVENTAYTAERQPERDITPVNDETMQEINTLLIALDKTWDDDLLP LCSQIFRRDIRASSELTQAEAVKALGFLKQKAAEQKVAA" CDS 9337..10017 /codon_start=1 /gene="exo" /product="5' to 3' double-stranded DNA exonuclease in the lambda Red system" /note="产生3'单链DNA尾部" /translation="MTPDIILQRTGIDVRAVEQGDDAWHKLRLGVITASEVHNVIAKPR SGKKWPDMKMSYFHTLLAEVCTGVAPEVNAKALAWGKQYENDARTLFEFTSGVNVTESP IIYRDESMRTACSPDGLCSDGNGLELKCPFTSRDFMKFRLGGFEAIKSAYMAQVQYSMW VTRKNAWYFANYDPRMKREGLHYVVIERDEKYMASFDEIVPEFIEKMDEALAEIGFVFG EQWR" RecA 10041..11102 /codon_start=1 /gene="recA" /product="DNA repair protein from E. coli" /translation="MAIDENKQKALAAALGQIEKQFGKGSIMRLGEDRSMDVETISTGS LSLDIALGAGGLPMGRIVEIYGPESSGKTTLTLQVIAAAQREGKTCAFIDAEHALDPIY ARKLGVDIDNLLCSQPDTGEQALEICDALARSGAVDVIVVDSVAALTPKAEIEGEIGDS HMGLAARMMSQAMRKLAGNLKQSNTLLIFINQIRMKIGVMFGNPETTTGGNALKFYASV RLDIRRIGAVKEGENVVGSETRVKVVKNKIAAPFKQAEFQILYGEGINFYGELVDLGVK EKLIEKAGAWYSYKGEKIGQGKANATAWLKDNPETAKEIEKKVRELLLSNPNSTPDFSV DDSEGVAETNEDF"
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