Template:SBB10 ConstructionFiles MaziarB sbb19

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 Construction of FokI Cleavage Domain basic part sbb19
 PCR mb0001/mb0003 on BBa_K157000 (504 bp, gp = A)
 PCR mb0002/mb0004 on BBa_K157000 (143 bp, gp = B)
 ---------------------------------------------------
 PCR mb0001/mb0004 on A+B           (622 bp, EcoRI/BamHI)
 Digest pBcajk2741-Bca1144          (EcoRI/BamHI, 2170+910, L)
 Product is pBjk2741-sbb19	      {<FokI+!}
 ---------------------------------------------------
 mb0001  Forward EcoRI + BglII 
ctacgGAATTCgctAGATCTCAGCTGGTTaaatctgaactggaggag
 mb0002  Forward point mutation A-->T  cataaaaccaatTgcaatggcgccg
 mb0003  Reverse point mutation A-->T  cggcgccattgcAattggttttatg
 mb0004  Reverse BamHI  gtacgGGATCCTTAaaaattgatctccccattg

JCA Notes

  • Fixed wiki link
  • Prefix should be ATG not GCT
  • Changed template name to partName
  • Construction file is correct otherwise
  • Changing some oligos for sharing with another part
 Construction of FokI Cleavage Domain basic part sbb19
 PCR fk0001/fk0003 on BBa_K243000 (504 bp, gp = A)
 PCR fk0004/mb0002 on BBa_K243000 (144 bp, gp = B)
 ---------------------------------------------------
 PCR fk0001/mb0002 on A+B           (622 bp, EcoRI/BamHI)
 Digest pBjk2741-Bca1144          (EcoRI/BamHI, 2170+910, L)
 Product is pBjk2741-sbb19	      {<FokI+!}
 ---------------------------------------------------
fk0001   Forward EcoRI and BglII for FokI 	 
ccaaaGAATTCatgAGATCTCAGCTGGTTaaatctgaactggaggag  
fk0004   (F)Base Substitution FokI   
ccataaaaccaatTgcaatggcgccg  
fk0003   (R)Base Substitution FokI   
cggcgccattgcAattggttttatgg
mb0002  Reverse BamHI  
gtacgGGATCCTTAaaaattgatctccccattg
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