pGL3 Luciferase Reporter Vectors Technical Manual #TM033

VIII.I. pGL3-Promoter Vector Restriction Sites (continued)Table 10. Restriction Enzymes That Cut the pGL3-Promoter Vector Between 1 and 5Times (continued).Enzyme # of Sites LocationKpnI 1 5MluI 1 15NaeI 3 1951, 2322, 4391NarI 1 313NcoI 1 278NgoMIV 3 1949, 2320, 4389NheI 1 21NotI 1 4843NspI 2 943, 2456PaeR7I 2 1867, 4458PpuMI 1 1459PshAI 1 2267Psp5II 1 1459PspAI 1 26PvuI 2 3715, 4761SacI 1 11SalI 1 2202Enzyme # of Sites LocationScaI 3 445, 3825, 4908SfiI 1 182SgrAI 1 1708SinI 3 1459, 3483, 3705SmaI 1 28SphI 1 943SrfI 1 28SspI 3 4149, 4702, 4817StuI 1 228StyI 2 229, 278VspI 1 3517XbaI 1 1934XcmI 1 1015XhoI* 1 32XmaI 1 26XmnI 1 3944*Due to the extent of supercoiling in this vector, the XhoI site has proven difficult to cut tocompletion under standard restriction digest conditions. For single XhoI digests, werecommend digesting the vector for a minimum of 2 hours using 20 units of enzyme permicrogram of DNA at 37°C to ensure complete digestion. If performing a double digestwith XhoI and another enzyme, linearize the vector using the companion enzyme prior tocarrying out the XhoI digest. Under these conditions, XhoI will cut the vector followingstandard reactions conditions.Table 11. Restriction Enzymes That Do Not Cut the pGL3-Promoter Vector.AatIIAccB7IAflIIAgeIApaIAscIBalIBbrPIBlpIBpu1102IBsp120IBssHIIBst1107IBst98IBstEIIBstXIBsu36ICspIEco72IEco81IEcoRIEcoRVI-PpoINdeINruINsiIPacIPflMIPinAIPmeIPmlIPpu10IPstIPvuIIRsrIISacIISgfISnaBISpeISplISse8387ISwaITth111IPromega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USAToll Free in USA 800-356-9526 · Phone 608-274-4330 · Fax 608-277-2516 · www.promega.comPart# TM033Printed in USA.Page 24 Revised 9/07