Thermo Fisher, K253520, pENTR™/TEV/D-TOPO™ Cloning Kit, with One Shot™ Mach1™-T1^R 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 with choice of vectors
• pENTR/TEV/D-TOPO vector includes Tobacco Etch Virus (TEV) recognition site for protease-dependent cleavage of an N-terminal tag from recombinant protein
• Vector has M13 and T7 primer sequencing sites and attL recombination sites flanking PCR product insertion site, allowing clones to be easily sequence verified and recombined into choice of attR containing Gateway destination 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