Please give a detailed
Here is a map for a typical DNA expression vector: CMV promoter: bases 232-819 T7 promoter/priming site: bases 863-882 Multiple cloning site: bases 895-1010 pcDNA3.1/BGH reverse priming site: bases 1022-1039 BGH polyadenylation sequence: bases 1028-1252 fl origin: bases 1298-1726 SV40 early promoter and origin: bases 1731-2074 Neomycin resistance gene (ORF): bases 2136-2930 SV40 early polyadenylation signal: bases 3104-3234 pUC origin: bases 3617-4287 (complementary strand) Ampicillin resistance gene (bla): bases 4432-5428 (complementary strand) ORF: bases 4432-5292 (complementary strand) Ribosome binding site: bases 5300-5304 (complementary strand) bla promoter (P3): bases 5327-5333 (complementary strand) a. You bought this vector from Invitrogen and will transform competent bacteria to grow up a stock of the plasmid. After bacterial transformation, what will you use to select for bacteria harboring the plasmid? b. Now that you have the purified plasmid, you want to linearize the plasmid and run it on an agarose gel to verify the size. Which restriction enzyme could you use to linearize the plasmid? c. You have cloned an insert into this vector and would like to make stable transfectants in your human immortalized fibroblast cell line. After transfection with this plasmid, what will you use to select for stable transfectants in the fibroblast culture?