Describe a design of an expression cassette and plasmid suitable for expression of a mammalian IgG in Pichia pastoris and the approach you would take to generate a high producer clonal cell line suitable for biomanufacturing of a pharmaceutical grade protein. Illustrate where possible alternative strategies, discuss these and justify your choice.

The answer would describe the design of the construct: optimizing the ribosome binding site (Kozak consensus sequence), codon optimizing the coding sequence, addition of restriction sites to enable cloning at a unique site downstream of the constitutive promoter sequence in the vector. I need to include the expression cassete design as well as vector