Diagrammatic representation of all-PCR cloning methodology.

The vector (pRS-NHS, -CHS, or -NOHS) contains two SmaI sites between ori (green box) and bla (brown box). Upon digestion with SmaI, two fragments are obtained that are joined together by PCR by means of complementary sequences (shown in red and blue) present both in the vector and in the gene (yellow box). In the control reaction, these fragments will not come together and join since insert is absent.