Table 1.

Gene-specific primers used for generating hSMVT truncations

Construct (Amino Acid)	Forward and Reverse Primers (5′–3′)	Positions, bp	Fragment, bp
hSMVT-GFP	CCGGAATTCATGAGTGTAGGGGTGAGC;	1-1905	1905
	ACGCGTCGACCAGGGAGGTCTCCTGGAG
hSMVT[1-632]-GFP	CCGGAATTCATGAGTGTAGGGGTGAGC;	1-1896	1896
	ACGCGTCGACCTCCTGGAGGATGCAGGT
hSMVT[1-624]-GFP	CCGGAATTCATGAGTGTAGGGGTGAGC;	1-1872	1872
	ACGCGTCGACGCTCCCCTGATAGGCTGTG
hSMVT[1-620]-GFP	CCGGAATTCATGAGTGTAGGGGTGAGC;	1-1860	1860
	ACGCGTCGACGGCTGCGGCAGCCAGGG
hSMVT[1-616]-GFP	CCGGAATTCATGAGTGTAGGGGTGAGC;	1-1848	1848
	ACGCGTCGACCGCGGCCATGGCCTCCT
hSMVT[1-612]-GFP	CCGGAATTCATGAGTGTAGGGGTGAGC;	1-1836	1836
	ACGCGTCGACCTCCTTGTCTCTGCTGTCCC
hSMVT[1-600]-GFP	CCGGAATTCATGAGTGTAGGGGTGAGC;	1-1800	1800
	ACGCGTCGACCGGCTTCTCAGGAAACAGGC
hSMVT[1-584]-GFP	CCGGAATTCATGAGTGTAGGGGTGAGC;	1-1752	1752
	ACGCGTCGACCCTGCAGTGGAGCCGCT
hSMVT[1-575]-GFP	CCGGAATTCATGAGTGTAGGGGTGAGC;	1-1725	1725
	ACGCGTCGACCAACGGAAGGAGGGACA
hSMVT[1-570]-GFP	CCGGAATTCATGAGTGTAGGGGTGAGC;	1-1710	1710
	ACGCGTCGACCAGGAGCTTTGGCAACACTG
hSMVT[1-567]-GFP	CCGGAATTCATGAGTGTAGGGGTGAGC;	1-1701	1701
	ACGCGTCGACTGGCAACACTGGGTAAAGG
hSMVT[ΔC]-GFP	CCGGAATTCATGAGTGTAGGGGTGAGC;	1-1656	1656
	ACGCGTCGACCATTCTCCCAGTGAGTAGACT

Shown is the combination of primer sequence used to generate full-length and truncated human sodium-dependent multivitamin transporter (hSMVT) constructs by PCR. Restriction sites for EcoRI (underlined) and Sal I (boldface) were added to the hSMVT primers to allow subsequent subcloning into the enhanced green fluorescent protein (GFP-N3) vector. hSMVT[ΔC]-truncation of COOH terminus of hSMVT.