How does the CACC genomic overhang work in a pentr plasmid?

Answer 1

The CACC genomic overhang is a method used in cloning with pENTR plasmids, which facilitate the recombination of DNA fragments into expression vectors. This technique involves adding a specific sequence of nucleotides (CACC) to the 5' end of the PCR product, enabling efficient insertion into the pENTR vector through Gateway cloning. The CACC overhang promotes directional cloning by ensuring that the insert is oriented correctly within the vector. This allows for streamlined downstream applications, such as protein expression and functional studies.