Patent ID: 9644211
Date: 2017-05-09
CPC Classifications: A61K,C12N

Claim:
1. A plasmid comprising the following units: (a) an endonuclease restriction site, (b) a promotorless endonuclease expression cassette, (c) a promoter that is not operably linked to the endonuclease expression cassette, (d) a sequence coding for an enzyme that catalyzes site-specific recombination, (e) at least two recognition sequences for the enzyme catalyzing site-specific recombination, (f) a sequence of interest between the at least two recognition sequences, and (g) an element preventing the expression of the endonuclease positioned between the at least two recognition sequences, wherein the promoter is operably linked to the endonuclease expression cassette after site-specific recombination occurs at the recognition sequences, and wherein, upon expression of the sequence coding for the enzyme that catalyzes site-specific recombination, the plasmid is divided into (i) a minicircle comprising the sequence of interest and the element preventing the expression of the endonuclease and (ii) a miniplasmid comprising the endonuclease restriction site, the endonuclease expression cassette, the promoter, and the sequence coding for an enzyme that catalyzes site-specific recombination.