Invitrogen™ pENTR™/SD/D-TOPO™ Cloning Kit, with One Shot™ TOP10 Chemically Competent E. coli 

Description

• Blunt-end PCR products clone directionally into pENTR/SD/D-TOPO entry vector at greater than 90% efficiency, and include an upstream Shine-Dalgarno sequence for expression in prokaryotes
• Rapidly shuttle cloned genes between multiple vector systems
• Reliable performance
• Directional cloning vector for entry to Gateway System and optimized for prokaryotic expression
• For access to Gateway System, PCR amplify the gene of interest and add product straight to the provided topoisomerase charged pENTR/SD/D-TOPO vector, incubate 5 minutes, and transform the provided competent E. coli cells
• Resulting attL containing Gateway Entry clones are ready for efficient recombination with choice of Gateway Destination vectors
• pENTR/SD/D-TOPO vectorincludes T7 gene 10 translational enhancer and ribosome binding site (RBS) for optimal expression of native protein after recombination with prokaryotic Gateway destination vectors
• Can also use vector for expression in other hosts systems by recombination with different destination vectors
• 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 in E. coli
• Four base overhang on vector pairs with four base sequence designed into forward primer used in PCR reaction to provide directionality to the topoisomerase ligation reaction

Cloning, Gateway Cloning