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. 2020 Dec 18;11:610032. doi: 10.3389/fpls.2020.610032

FIGURE 1.

FIGURE 1

Schematic illustration of the cloned target vectors and the final GNN and CCSB constructs used for transfection. (A) pJET1.2-based assembly vector with a designed multiple cloning site for generation of the GNE, NANS and NANP-containing GNN construct with homologous flanks for integration into the adenine phosphoribosyltransferase (APT) locus. (B) pTOPO-based assembly vector with a designed multiple cloning site for generation of the CMAS, CSAT, ST, and BSD-containing CCSB construct with homologous flanks for integration into the prolyl-4-hydroxylase 1 (P4H1) locus. LguI or NotI were used to linearize GNN (C) and CCSB (D) constructs, respectively, before sequential transformation of Physcomitrella. 5′ HR, 5′ homologous region; 3′ HR, 3′ homologous region; 35S P long, long CaMV 35S promoter; 35S P, CaMV 35S promoter; nosT, nos terminator; 35ST, CaMV 35S terminator.