Minicircles are Next Generation Gene Vectors consisting almost exclusively of the gene of interest (GOI) and a very short sequence which inevitably remains in the Minicircle (SCAR). During Minicircle production intramolecular recombination in E. coli yields two circular DNA structures – the Minicircle and a miniplasmid. In comparison to typical plasmid DNA with the general portion known as the bacterial backbone, comprising the genetic marker for selection (e.g. by antibiotics) and the so-called origin of replication (ori; for the replication of the plasmids in E. coli cells), Minicircles are extremely small – down to half the size of the smallest plasmids. The dissemination of antibiotic resistance genes and the uncontrolled expression of backbone sequences may have a strongly negative influence on gene expression and could elicit immune response of the target cells. In addition, it was shown that unmethylated CpG motifs contribute to the silencing of episomal genes. In vector development, it is, therefore, an important objective to produce supercoiled (ccc) Minicircle DNA without bacterial sequences. The used production method leading to a highly homogeneous Minicircle product is proprietary to PlasmidFactory which produces both standard Minicircle (In Stock products) as well as customized Minicircle DNA.