Template:SBB10 ConstructionFiles PaulinaT sbb21

Construction of FokI+ cleavage domain basic part sbb21 PCR fk0001/fk0003 on FokI	(505 bp, gp = A) PCR fk0004/fk0002 on FokI	(141 bp, gp = B) - PCR fk0001/fk0002 on A+B          (620 bp, EcoRI/BamHI) Digest pBjk2741-Bca1144           (EcoRI/BamHI, 2170+910, L) Product is pBjk2741-ssb21          {} - fk0001  Forward EcoRI and BglII for FokI 	 ccaaaGAATTCgtccAGATCTCAGCTGGTTaaatctgaactggaggag fk0004  (F)Base Substitution FokI   ccataaaaccaatTgcaatggcgccg fk0003  (R)Base Substitution FokI   cggcgccattgcAattggttttatgg fk0002  Reverse BamHI for FokI	     gctagGGATCCaaaattgatctccccattgttg



JCA Notes
Construction of FokI+ cleavage domain basic part sbb21 PCR fk0001/fk0003 on K243000   (504 bp, gp = A) PCR fk0004/fk0002 on K243000    (141 bp, gp = B) - PCR fk0001/fk0002 on A+B          (619 bp, EcoRI/BamHI) Digest pBjk2741-Bca1144           (EcoRI/BamHI, 2170+910, L) Product is pBjk2741-ssb21          {} - fk0001  Forward EcoRI and BglII for FokI 	 ccaaaGAATTCatgAGATCTCAGCTGGTTaaatctgaactggaggag fk0004  (F)Base Substitution FokI   ccataaaaccaatTgcaatggcgccg fk0003  (R)Base Substitution FokI   cggcgccattgcAattggttttatgg fk0002  Reverse BamHI for FokI	     gctagGGATCCaaaattgatctccccattgttg
 * Correct assembly, prefix should be ATG not GTCC
 * Explicitly put in the name of the template plasmid, not just a description; ie, what the tube will have written on the cap