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pTCK303

Catalog Number P0273
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Transfection-Grade Plasmid QC Standards View Plasmid Manual
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Product Overview

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.

Product Disclaimer

  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:
    Product names and IDs are provided by the MiaoLing Plasmid Platform.
  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.”

Related Publications

No Literature Available

Product Documents

P0273_Sequencing report.zip

Plasmid Elements

intron 10..199 /note="蓖麻豆过氧化氢酶内含子，修改型" 6xHis 2024..2041 /codon_start=1 /product="6xHis affinity tag" /translation="HHHHHH" terminator 2076..2328 /note="农杆菌合成酶终止子和polyA信号" RB T-DNA repeat 2350..2374 /note="来自胭脂碱C58 T-DNA的右侧边界重复序列" CDS 3674..4303 /codon_start=1 /product="stability protein from the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI" bom 6408..6548 /note="源自pBR322的移动区域基础" ori complement(6734..7322) /direction=LEFT /note="高拷贝数 ColE1/pMB1/pBR322/pUC 复制起点" KanR complement(7409..8203) /codon_start=1 /gene="aphA-3" /product="aminoglycoside phosphotransferase" /note="赋予对卡那霉素的抗性" /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF" LB T-DNA repeat 8628..8652 /note="左端重复序列来自胭脂碱C58 T-DNA" CaMV poly(A) signal 8730..8904 /note="菜花花叶病毒聚腺苷酸信号" HygR complement(8944..9969) /codon_start=1 /gene="aph(4)-Ia" /product="aminoglycoside phosphotransferase from E. coli" /note="赋予对潮霉素的抗性" /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK K" CaMV 35S promoter (enhanced) complement(10036..10713) /note="菜花花叶病毒35S启动子，带有重复的增强子区域。" CAP binding site 10905..10926 /bound_moiety="E. coli catabolite activator protein" /note="CAP结合激活了cAMP存在的条件下的转录。" lac promoter 10941..10971 /note="大肠杆菌乳糖操纵子的启动子" lac operator 10979..10995 /bound_moiety="lac repressor encoded by lacI" /note="lac阻遏蛋白与lac操纵子结合以抑制大肠杆菌中的转录。这种抑制可以通过添加乳糖或异丙基-β-D-硫代半乳糖苷（IPTG）来解除。" primer_bind 11003..11019 /note="通用测序引物，多个类似变体之一" terminator 11036..11288 /note="农杆菌根瘤合成酶终止子和polyA信号" Ubi promoter complement(11853..13845) /note="maize polyubiquitin gene promoter" primer_bind complement(13861..13877) /note="common sequencing primer, one of multiple similar variants" CaMV 35S promoter 14254..14599 /note="strong constitutive promoter from cauliflower mosaic virus" GUS join(14616..14621,1..9,200..2017) /codon_start=1 /product="beta-glucuronidase" /note="这个版本的gusA基因具有一个带有过氧化氢酶内含子的5'延伸，以确保在植物中而不是细菌中表达。" /translation="MVDLRNRRLVRPVETPTREIKKLDGLWAFSLDRENCGIDQRWWES ALQESRAIAVPGSFNDQFADADIRNYAGNVWYQREVFIPKGWAGQRIVLRFDAVTHYGK VWVNNQEVMEHQGGYTPFEADVTPYVIAGKSVRITVCVNNELNWQTIPPGMVITDENGK KKQSYFHDFFNYAGIHRSVMLYTTPNTWVDDITVVTHVAQDCNHASVDWQVVANGDVSV ELRDADQQVVATGQGTSGTLQVVNPHLWQPGEGYLYELEVTAKSQTESDIYPLRVGIRS VAVKGQQFLINHKPFYFTGFGRHEDADLRGKGFDNVLMVHDHALMDWIGANSYRTSHYP YAEEMLDWADEHGIVVIDETAAVGFQLSLGIGFEAGNKPKELYSEEAVNGETQQAHLQA IKELIARDKNHPSVVMWSIANEPDTRPQGAREYFAPLAEATRKLDPTRPITCVNVMFCD AHTDTISDLFDVLCLNRYYGWYVQSGDLETAEKVLEKELLAWQEKLHQPIIITEYGVDT LAGLHSMYTDMWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNFADFATSQGILRVGGNKK GIFTRDRKPKSAAFLLQKRWTGMNFGEKPQQGGKQ"
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