Template:SBB10 ConstructionFiles AmyK sbb22

Construction of FokI- basic part PCR ak10F/ak20R    Registry part BBa_K243001            (230 bp, gp = A) PCR ak20F/ak30R     Registry part BBa_K243001            (71 bp, gp = B) PCR ak30F/ak40R     Registry part BBa_K243001            (245 bp, gp = C) PCR ak40F/ak11R     Registry part BBa_K243001            (135 bp, gp = D) --- PCR ak10F/ak11R on A+B+C+D          (619bp, EcoRI/BamHI) Digest pBjk2741-Bca1144             (EcoRI/BamHI, 910+2170, L) Product is pBjk2741-Bak1234  {} --- ak10F Forward EcoRI for FokI- CCATAgaattcCAGagatctCAGCTGGTTaaatctgaactggaggagaa ak20F Base substitution 1  gtaaaccagACggtgccatt ak20R Base substitution 1  aatggcaccGTctggtttac ak30F Base substitution 2  cgttatcgtggAcacaaaagcg ak30R	Base substitution 2 cgcttttgtgTccacgataacgc ak40F Base substitution 3  caccaatTgcaatggggcag ak40R Base substitution 3  ctgccccattgcAattggtg ak11R Reverse BamHI for FokI- CTGATggatccaaaattgatctcgccattg



JCA Notes
Construction of FokI- basic part PCR gh1000F/gh1001R on BBa_K243001  (261 bp, gp = A) PCR gh1001F/gh1003R on BBa_K243001   (264 bp, gp = B) PCR gh1003F/ak11R     Registry part BBa_K243001            (141 bp, gp = C) --- PCR gh1000F/ak11R on A+B+C          (619bp, EcoRI/BamHI) Digest pBjk2741-Bca1144             (EcoRI/BamHI, 910+2170, L) Product is pBjk2741-sbb22   {} --- gh1000F Forward for part <FokI-! ccagtGAATTCatgAGATCTCAGCTGGTTaaatctgaactggaggag gh1001F Making internal mutation 1+2 cgttggttccccgatcgattatggcgttatcgtggAcacaaaagc gh1001R Making internal mutation 1+2 cgccataatcgatcggggaaccaacggtataaatggcaccGTctggtttac gh1003F Making internal mutation 3   ccatatcaccaatTgcaatggggcagtgctgag gh1003R Making internal mutation 3   ccattgcAattggtgatatgg ak11R Reverse BamHI for FokI- CTGATggatccaaaattgatctcgccattg
 * Assembles fine
 * Changed seq on Clotho, restriction sites were included
 * Name of part should be sbb22, changed wikilink, and corrected file
 * Due to similarity to another part, shared some oligos