Thermo Fisher, K252520, pENTR™/TEV/D-TOPO™ Cloning Kit, with One Shot™ TOP10 Chemically Competent <i>E. coli</i>

Quantity: 20 Reactions
Unit Size: Each
• Blunt-end PCR products clone directionally into pENTR/TEV/D-TOPO entry vector at greater than 90% efficiency, and include TEV protease-dependent cleavage site for removal of N-terminal tags from expressed proteins
• Rapidly shuttle cloned genes between multiple vector systems
• Reliable performance
• For access to Gateway System, PCR amplify gene of interest and add product straight to provided topoisomerase charged pENTR/SD/D-TOPO vector, incubate 5 minutes, and transform provided competentE. colicells
• Resulting attL containing Gateway Entry clones are ready for efficient recombination
• Optimized pENTR/TEV/D-TOPO Vector includes Tobacco Etch Virus (TEV) recognition site for protease-dependent cleavage of N-terminal tag from recombinant protein
• Vector has M13 and T7 primer sequencing sites and attL recombination sites flanking the PCR product insertion site, allowing clones to be easily sequence-verified and recombined into choice of vectors
• Kanamycin resistance gene and pUC origin are used for selection and high-copy propagation inE. coli
• Four base overhang on vector pairs with four base sequence designed into forward primer used in PCR reaction to provide directionality to topoisomerase ligation reactionCloning, Gateway Cloning