Construction of pNGG. (A) Alignment of (from top to bottom): the 5' and 3' ends of pTH31 Gateway® cassette; XhoI-att R1 PCR primer; BamHI-att R2 PCR primer. Sequence identity is denoted by asterisks below the alignment. (B) The Gateway® cassette was PCR amplified using pTH31 as template, and either primer XhoI-att R1 alone (lane 1), primer BamHI-att R2 alone (lane 2), or primers XhoI-att R1 and BamHI-att R2 (lane 3). Markers size is indicated on the left in base pairs. (C) "Two-halves" making of pNGG. The plasmids are not at scale. Light grey, Gateway cassette. Black, att R recombination sites. Primer (1), XhoI-att R1. Primer (2), BamHI-mut-R. Primer (3), BamHI-mut-F. Primer (4), BamHI-att R2 (Table 3).