TABLE 1.

Primer pairs for assembly of the TGEV infectious construct^a

Primer	Sequence	Position in genome	Orientation	Purpose
TGEV 5′ T7	5′-GTCGGCCTCTTAATACGACTCACTATAGACTTTTAAAGTAAAGTGAG-3′	1–19	+	Insert T7 start TGEV A, 5′ end
TGEV A 3,642	5′-TGTTGAAGAAATCAAAGGCCTG-3′	3621–3642	−	Remove T7 stop
TGEV A 3,621	5′-CAGGCCTTTGATTTCTTCAAC-3′	3621–3641	+	Remove T7 stop, overlap RT-PCR
	T
TGEV A 6,180	5′-CTTGGAGATGTTGAAAATCAGC-3′	6180–6201	−	TGEV A, 3′ end
TGEV B1 6,134	5′-TAAAGTCTGCAGTCTGTGGC-3′	6115–6134	+	TGEV B1, 5′ end
TGEV B1 9,957	5′-TAATCCAAGTGAATGGTGTGTAC-3′	9935–9957	−	TGEV B1, 3′ end, insert BstXI site
TGEV B2 9,939	5′-ACACCATTCACTTGGATTAATCC-3′	9939–9961	+	TGEV B2, 5′ end, insert BstXI site
	C
TGEV B2 11,342	5′-GTAGCCGATGCGGCTGGAATG-3′	11342–11362	−	TGEV B2, 3′ end
TGEV C 11,345	5′-TCCAGCCGCATCGGCTACAAG-3′	11345–11365	+	TGEV C, 5′ end, insert BglI site
	T     A
TGEV C 13,605	5′-GTGCAAAGAAGAAGTGTTTTAATG-3′	13605–13628	−	Remove T7 stop
TGEV C 13,609	5′-AAAACACTTCTTCTTTGCACAGG-3′	13609–13631	+	Remove T7 stop, overlap RT-PCR
	T  T
TGEV C 16,580	5′-TGTGCCAAGAAGGCCTTGACAAC-3′	16580–16602	−	TGEV C, 3′ end
TGEV DE 16,585	5′-CAAGGCCTTCTTGGCACATAATC-3′	16585–16607	+	TGEV DE, 5′ end, insert BglI site
	T  A
TGEV DE 3,133	5′-GTCTAGCCTGCACGGCTACTGC-3′	23475–23496	−	TGEV DE, 3′ end
TGEV F 3,112	5′-GCAGTAGCCGTGCAGGCTAGAC	23475–23496	+	TGEV F, 5′ end, insert BglI site
	A  T
TGEV 3′ end	5′-NNNNNNGCGGCCGCTTTTTTTTTTTTTTTTTTTTTTTTTGGTGTATCACTATCAAAAGG-3′	3′ end, poly(A) tail	−	TGEV F 3′ end, Poly(A) tail, NotI

^aWhen a primer sequence differs from the wild-type sequence, the wild-type nucleotide is shown below the sequence. Restriction sites are shown in boldface, and the T7 start site in underlined. Sequences are from references 3, 11, 17, and 33.