Cloning from genomic DNA

Design your primers with 18-19 bases of overlap with sequence to be cloned. Then add restriction enzyme sites and any extra residues wanted (eg His-tag) to this.

Check that your gene will be in-frame after inclusion into your vector using the restriction sites you have chosen.

Check the vector has a ribosome binding site - you may need to use the intrinsic RBS from the gene (if an E. coli gene, otherwise design a suitable one). The RBS is a tract of 5x purine bases ~50bps upstream of the start methionine. You can find the biological RBS for any E. coli gene by searching for the gene in NCBI database and then manually changing the boundaries until you find the RBS.

PCR up your fragment and gel extract