Template:SBB10 ConstructionFiles JimH sbb13

2) sbb13: PhiC31 Integrase

PCR OJIMH011/OJIMH012 on pBca9523-Bca1623	(949 bp, gp = A) PCR OJIMH013/OJIMH014 on pBca9523-Bca1659	(938 bp, gp = B) - PCR PJIMH011/OJIMH014 on A+B	(1843 bp, EcoRI/BamHI) Sub in pBjk2741-Bca1144	(EcoRI/BamHI, 910+2170, L) Product is pBjk2741-sbb13	{} - OJIMH011	Forward PCR of Part 1 of PhiC31	ccataGAATTCatgAGATCTGACACGTACGCGGGTGCTTA OJIMH012	SOEing of Part 1 (R) of PhiC31	AAGAAGCCGGACGGCACGCCGACCacgaagattgagggttaccg OJIMH013	SOEing of Part 2 (F) of PhiC31	cggtaaccctcaatcttcgtGGTCGGCGTGCCGTCCGGCTTCTT OJIMH014	Reverse PCR of Part 2 of PhiC31	atcagGGATCCCGCCGCTACGTCTTCCGT



JCA Notes

 * Oligos 12 and 13 are switched
 * Otherwise correct

sbb13: PhiC31 Integrase

PCR OJIMH011/OJIMH013 on Bca1623 6-9           (949bp, gp=A) PCR OJIMH012/CA1674 on Bca1559 7-7             (514bp, gp=B) PCR CA1675/OJIMH014 on Bca1559 7-1	       (444bp, gp = C) - PCR PJIMH011/OJIMH014 on A+B+C	(1843 bp, EcoRI/BamHI) Sub in pBjk2741-Bca1144	(EcoRI/BamHI, 910+2170, L) Product is pBjk2741-sbb13	{} - OJIMH011	Forward PCR of Part 1 of PhiC31 ccataGAATTCatgAGATCTGACACGTACGCGGGTGCTTA OJIMH012	SOEing of Part 1 (R) of PhiC31 AAGAAGCCGGACGGCACGCCGACCacgaagattgagggttaccg OJIMH013	SOEing of Part 2 (F) of PhiC31 cggtaaccctcaatcttcgtGGTCGGCGTGCCGTCCGGCTTCTT CA1674	GGGCGTCGGCGCGCTCCGCAACAAGGTTCGCCCGTTCGCCGCTCTTCTCAG PCA assembly of phiCthreeprime (Bca1659) CA1675	TGCGGAGCGCGCCGACGCCCTGAACGCCCTTGAAGAGCTGTACGAAGACCG PCA assembly of phiCthreeprime (Bca1659) OJIMH014	Reverse PCR of Part 2 of PhiC31 atcagGGATCCCGCCGCTACGTCTTCCGT