Restriction digests of the clone give the following sizes (kb): BamHI--5.2, 1.4; PstI--6.0, 0.8; XbaI--3.6, 3.0. Insert contains the following restriction sites (approximate kb from the 5' end): BamHI--1.95, 3.26; BglII--2.19; EcoRI--1.17; EcoRV--0.69; HindIII--1.99, 2.48; KpnI--0.89, 1.18, 2.76; NcoI--0.03, 1.43; PstI--2.74; PvuII--2.30, 2.72; SacI--0.59; SmaI--0.45, 2.29; XbaI--0.05; XhoI--0.14. Insert contains the complete coding sequence for ORFs 2-5 and includes approximately 30 bp polyadenylation. Virus was propagated in Nicotiana benthamiana. |