Table 1. hNTAN1 constructs and their relevant expression characteristics in BL21 (DE3).

The sequences of the sense and antisense primers used for the modified termini and RBS are found in Supplementary Table 1. The resultant amino acid sequences are shown where residues following the ellipsis are indicative of the C-terminus of the sequence. The purity of hNTAN1 was evaluated by SDS-PAGE, where band intensities were estimated by densitometry imaging (Quantity 1, BioRad).

Plasmid	RBS	Amino Acid Sequence	hNTAN1 Purity
pHisNTAN*	AAGGAGA	MGGSSHHHHHHSSGPLLV…SGPS	30 %
pNTANHis	AAGGAGA	MGPLLV…SGPSGGSSHHHHHH	30 %
pR1NTANSII	AAGGAGA	MGPLLV…SPGSSAWSHPQFEK	30 %
pR2NTANSII	AACAATAATAAGGAGATAAGAA	MGPLLV…SPGSSAWSHPQFEK	70 %
pNTANFLAGSII	AACAATAATAAGGAGATAAGAA	MGPLLV…SPGSGGGSDYKDDDDKSAWSHPQFEK	> 98%
pNTAN2FLAGSII	AACAATAATAAGGAGATAAGAA	MPLLV…SPGSGGGSDYKDDDDKSAWSHPQFEK	> 98%

^*Trace activity detected for His₆-hNTAN1 through addition of final purified product (10μg total protein) to 1mM N¹-AII for 90 min at 37°C in 50mM Tris-HCl, 150mM NaCl, pH 7.5. Reactions were quenched with 12% TCA (w/v) and then evaluated by HPLC for the presence of a peak corresponding to the expected product AII. For all other plasmids, the presence of hNTAN1 activity could be detected through the addition of final purified product (10–100ng total protein) to 100μM N¹-AII for 1 min at 37°C in the same buffer and evaluated similarly.