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. 2021 Oct 28;22(21):11641. doi: 10.3390/ijms222111641

Figure 1.

Figure 1

Construction of SARS-CoV-2 subreplicon RNA expression vector. (A) Schematic structure of SARS-CoV-2 genomic RNA and SARS-CoV-2 subreplicon RNA. SARS-CoV-2 subreplicon RNA expression vector contains 5′-UTR (1–265), a part of ORF1a (266–475, green box), nucleocapsid (N, 28277–29530, blue box), and ORF10 (open box) and 3′-UTR (29531–29870), and the firefly luciferase sequence was inserted between ORF1a and nucleocapsid. The secondary structure of SARS-CoV-2 genomic RNA presented by Wacker et al. is schematically shown [23], and the protein translated regions of ORF1a, luciferase, and nucleocapsid are boxed and colored with green, yellow, and blue, respectively. Spike (S), membrane (M), and nucleocapsid (N) regions are boxed and marked S, M, and N, respectively. SARS-CoV-2 subreplicon RNA is numbered based on the Gene Bank MN994467 SARS-CoV-2 genomic RNA sequence. (B) SARS-CoV-2 subreplicon cDNA was inserted into the CMV promoter (orange arrow) for construction of the expression vector. Lengths of SARS-CoV-2 subreplicon cDNA regions are numbered. SARS-CoV-2 subreplicon RNA is expressed with poly-A tail (purple box). A fusion protein of part of ORF1a, firefly luciferase, and nucleocapsid is translated from the mRNA.