From OpenWetWare
2) sbb14: PhiC31 Integrase
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PCR AS009-F/AS010-R on pBca9523-Bca1623 (980 bp, gp = A)
PCR AS011-F/AS012-R on pBca9523-Bca1659 (938 bp, gp = B)
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PCR AS009-F/AS010-R on A+B (1874 bp, EcoRI/BamHI)
Sub in pBjk2741-Bca1144 (EcoRI/BamHI, 910+2170, L)
Product is pBjk2741-sbb14 {rbs.phiC31>}
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AS009-F Forward PCR of Part 1 of PhiC31 with RBS ccataGAATTCatgAGATCTACATACCGAGATCATATATAAGGAGGTACATatgGACACGTACGCGGGTGCTTACG
AS010-R SOEing of Part 1 of PhiC31 cggtaaccctcaatcttcgtGGTCGGCGTGCCGTCCGGCTTCTT
AS011-F SOEing of Part 2 of PhiC31 AAGAAGCCGGACGGCACGCCGACCacgaagattgagggttaccg
AS012-R Reverse PCR of Part 2 of PhiC31 atcagGGATCCCGCCGCTACGTCTTCCGT
JCA Notes
- BglII site missing from the first oligo
- Otherwise correct
- Need to put part into Clotho
sbb14: PhiC31 Integrase
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PCR AS009-F/AS010-R on Bca1623 6-9 (976bp, gp=A)
PCR AS011-F/CA1674 on Bca1559 7-7 (514bp, gp=B)
PCR CA1675/AS012-R on Bca1559 7-1 (444bp, gp=C)
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PCR AS009-F/AS012-R on A+B+C (1868 bp, EcoRI/BamHI)
Sub in pBjk2741-Bca1144 (EcoRI/BamHI, 910+2170, L)
Product is pBjk2741-sbb14 {rbs.phiC31>}
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AS009-F Forward PCR of Part 1 of PhiC31 with RBS
ccataGAATTCatgAGATCTCAGATCATATATAAGGAGGTACATatgGACACGTACGCGGGTGCTTACG
AS010-R SOEing of Part 1 of PhiC31
cggtaaccctcaatcttcgtGGTCGGCGTGCCGTCCGGCTTCTT
AS011-F SOEing of Part 2 of PhiC31
AAGAAGCCGGACGGCACGCCGACCacgaagattgagggttaccg
CA1674 GGGCGTCGGCGCGCTCCGCAACAAGGTTCGCCCGTTCGCCGCTCTTCTCAG
PCA assembly of phiCthreeprime (Bca1659)
CA1675 TGCGGAGCGCGCCGACGCCCTGAACGCCCTTGAAGAGCTGTACGAAGACCG
PCA assembly of phiCthreeprime (Bca1659)
AS012-R Reverse PCR of Part 2 of PhiC31
atcagGGATCCCGCCGCTACGTCTTCCGT
Part: GATCTCAGATCATATATAAGGAGGTACATatgGACACGTACGCGGGTGCTTACGACCGTCAGTCGCGCGAGCGCGAAAATTCGAGCGCAGCAAGCCCAGCGACACAGCGTAGCGCCAACGAAGACAAGGCGGCCGACCTCCAGCGCGAAGTCGAGCGCGACGGGGGCCGGTTCAGGTTCGTCGGGCATTTCAGCGAAGCGCCGGGCACGTCGGCGTTCGGGACGGCGGAGCGCCCGGAGTTCGAACGCATCCTGAACGAATGCCGCGCCGGGCGGCTCAACATGATCATTGTCTATGACGTGTCGCGCTTCTCGCGCCTCAAGGTCATGGACGCGATTCCGATTGTCTCGGAATTGCTCGCCCTGGGCGTGACGATTGTTTCCACTCAGGAAGGCGTCTTCCGGCAGGGAAACGTCATGGACCTGATTCACCTGATTATGCGGCTCGACGCGTCGCACAAAGAATCTTCGCTCAAGTCGGCGAAGATTCTCGACACGAAGAACCTCCAGCGCGAATTGGGCGGGTACGTCGGCGGGAAGGCGCCTTACGGCTTCGAGCTTGTTTCGGAGACGAAGGAGATCACGCGCAACGGCCGAATGGTCAATGTCGTCATCAACAAGCTTGCGCACTCGACCACTCCCCTTACCGGACCCTTCGAGTTCGAGCCCGACGTAATCCGGTGGTGGTGGCGTGAGATCAAGACGCACAAACACCTTCCCTTCAAGCCGGGCAGTCAAGCCGCCATTCACCCGGGCAGCATCACGGGGCTTTGTAAGCGCATGGACGCTGACGCCGTGCCGACCCGGGGCGAGACGATTGGGAAGAAGACCGCTTCAAGCGCCTGGGACCCGGCAACCGTTATGCGAATCCTTCGGGACCCGCGTATTGCGGGCTTCGCCGCTGAGGTGATCTACAAGAAGAAGCCGGACGGCACGCCGACCacgaagattgagggttaccgCATTCAGCGCGACCCGATCACGCTCCGGCCGGTCGAGCTTGATTGCGGACCGATCATCGAGCCCGCTGAGTGGTATGAGCTCCAGGCGTGGTTGGACGGCAGGGGGCGCGGCAAGGGGCTTTCCCGGGGGCAAGCCATTCTGTCCGCCATGGACAAGCTGTACTGCGAGTGTGGCGCCGTCATGACTTCGAAGCGCGGGGAAGAATCGATCAAGGACTCTTACCGCTGCCGTCGCCGGAAGGTGGTCGACCCGTCCGCACCTGGGCAGCACGAAGGCACGTGCAACGTCAGCATGGCGGCACTCGACAAGTTCGTTGCGGAACGCATCTTCAACAAGATCAGGCACGCCGAAGGCGACGAAGAGACGTTGGCGCTTCTGTGGGAAGCCGCCCGACGCTTCGGCAAGCTCACTGAGGCGCCTGAGAAGAGCGGCGAACGGGCGAACCTTGTTGCGGAGCGCGCCGACGCCCTGAACGCCCTTGAAGAGCTGTACGAAGACCGCGCGGCAGGCGCGTACGACGGACCCGTTGGCAGGAAGCACTTCCGGAAGCAACAGGCAGCGCTGACGCTCCGGCAGCAAGGGGCGGAAGAGCGGCTTGCCGAACTTGAAGCCGCCGAAGCCCCGAAGCTTCCCCTTGACCAATGGTTCCCCGAAGACGCCGACGCTGACCCGACCGGCCCTAAGTCGTGGTGGGGGCGCGCGTCAGTAGACGACAAGCGCGTGTTCGTCGGGCTCTTCGTAGACAAGATCGTTGTCACGAAGTCGACTACGGGCAGGGGGCAGGGAACGCCCATCGAGAAGCGCGCTTCGATCACGTGGGCGAAGCCGCCGACCGACGACGACGAAGACGACGCCCAGGACGGCACGGAAGACGTAGCGGCGG
