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The transformation of cDNA into plant is still not carried out directly by a universal transgenic vector, hence it is not convenient for the transgenic application of those function-important cDNAs. Herein we introduced a rapid, cost-effective PCR-based DNA synthesis method to construct a new universal plant overexpression vector for the direct inserting of target cDNA during transgenic application. The new engineered transgenic vector was based on the pCAMBIA vector backbone, which was inserted by five motifs, including a promoter, an enhancer, a Linker DNA fragment, a endoplasmic reticulum retention signal (KDEL), and a NOS-T DNA. The target cDNA can be directly inserted into the new universal vector to construct the workable transgenic plasmid according to the interesting need of transgenic users. The result reported here will promote the plant transgenic application of cDNAs.