Table 1.

Specific activities of AloI, PpiI, TstI, and hybrid REases

Enzyme	Recognition sequence*	Specific activity†
AloI	↓(7/12–13)GGAN6GTTC(12–13/7)↓	4,700
PpiI‡	↓(8/13–14)GAGN5GTTC(12/7)↓	18,600
TstI	↓(7/12)GGAN6GTG(13/8)↓	18,900
Alo-PpiSR	↓(8/13–14)GAGN5GTTC(12–13/7)↓	4,600
Alo-PpiTRD1	↓(8/13–14)GAGN5GTTC(12–13/7)↓	4,600
Ppi-AloSR	↓(7/12–13)GGAN6GTTC(12/7)↓	9,500
Ppi-AloTRD1	↓(7/12–13)GGAN6GTTC(12/7)↓	18,900
Tst-PpiTRD1-Gly1006Leu§	↓(8/14–15)GAGN5GTG(13–14/8)↓	<1,500

*Purification and cleavage position determination of hybrid proteins is described in Methods in http://www.pnas.org/cgi/content/full//DC1SI Text.

^†Specific activity is given as units per milligram of enzyme. One unit is defined as the amount of enzyme required to cleave 1 μg of BamHI-linearized DNA of pSEAd-7 in 1 h at optimal temperature in a reaction volume of 50 μl.

^‡Cleavage positions may vary depending on the DNA environment.

^§Detailed description of cleavage position determination is provided in Cleavage Positions of the New Specificity REase in SI Text and SI Fig. 7.