I want to over-express one operon within a bacterium. The operon has about 10 genes, each 1 kb in size. How can I design the expression system?
The following paper describes a technique for capturing and expressing tens of kb gene clusters. You could try this technique.
http://www.pnas.org/content/111/5/1957.long
Otherwise you could try traditional PCR based cloning and sequencing to ensure no errors, but you are on the upper limit for this technique.
You can also try yeast recombination approach to assemble the operon, which requires very minimum amounts of overlaps to capture by yeast and assembled with a digested vector.
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