Template:SBB10 ConstructionFiles DanielS sbb10
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PCR DS001 and DS002 on pBca9523-Bca1587 (1051 bp, EcoRI/BamHI) Sub into pBjk2741 (EcoRI/BamHI, 2170+910, L) Product is pBjk2741-sb10 {<SB100x!} ---------------------------- DS001 Forward oligo for cloning of <SB100x! CCATAgaattcTCCagatctGGTAAATCTAAAGAAATCTCTCAGGAC DS002 Reverse oligo for cloning of <SB100x! ctgatGGATCCttaGTATTTGGTAGCGTTACCTT
JCA Notes
- Prefix region should be atg, not TCC
- Otherwise correct
- Need to put part on Clotho
- Changed to SOEing due to template errors
Construction of sbb10 PCR DS001/CA1600 on Bca1587 5-1 (420bp, gp=A) PCR CA1597/DS002 on Bca1587 5-6 (767bp, gp=B) ---------------------------- PCR DS001 and DS002 on A+B (1051 bp, EcoRI/BamHI) Sub into pBjk2741 (EcoRI/BamHI, 2170+910, L) Product is pBjk2741-sb10 {<SB100x!} ---------------------------- DS001 Forward oligo for cloning of <SB100x! CCATAgaattcATGagatctGGTAAATCTAAAGAAATCTCTCAGGAC CA1600 CGAAACGCAGACGAGCTTTTTTGTGACGGTTCTGGAGCAGCGGTTTTTT PCA assembly of sleepingBeauty (Bca1587) CA1597 ATGCTGGAAGAAACCGGTACCAAAGTTTCTATCTCTACCGTTAAACGTG PCA assembly of sleepingBeauty (Bca1587) DS002 Reverse oligo for cloning of <SB100x! ctgatGGATCCttaGTATTTGGTAGCGTTACCTT Part: gatctGGTAAATCTAAAGAAATCTCTCAGGACCTGCGTAAACGTATCGTTGACCTGCACAAATCTGGTTCTTCTCTGGGTGCTATCTCTAAACGTCTGGCTGTTCCGCGTTCTTCTGTTCAGACCATCGTTCGTAAATACAAACACCACGGTACCACCCAGCCGTCTTACCGTTCTGGTCGTCGTCGTGTTCTGTCTCCGCGTGACGAACGTACCCTGGTTCGTAAAGTTCAGATCAACCCGCGTACCACCGCTAAAGACCTGGTTAAAATGCTGGAAGAAACCGGTACCAAAGTTTCTATCTCTACCGTTAAACGTGTTCTGTACCGTCACAACCTGAAAGGTCACTCTGCTCGTAAAAAACCGCTGCTCCAGAACCGTCACAAAAAAGCTCGTCTGCGTTTCGCTACCGCTCACGGTGACAAAGACCGTACCTTCTGGCGTAACGTTCTGTGGTCTGACGAAACCAAAATCGAACTGTTCGGTCACAACGACCACCGTTACGTTTGGCGTAAAAAAGGTGAAGCTTGCAAACCGAAAAACACCATCCCGACCGTTAAACACGGTGGTGGTTCTATCATGCTGTGGGGTTGCTTCGCTGCTGGTGGTACCGGTGCTCTGCACAAAATCGACGGTATCATGGACGCTGTTCAGTACGTTGACATCCTGAAACAGCACCTGAAAACCTCTGTTCGTAAACTGAAACTGGGTCGTAAATGGGTTTTCCAGCACGACAACGACCCGAAACACACCTCTAAAGTTGTTGCTAAATGGCTGAAAGACAACAAAGTTAAAGTTCTGGAATGGCCGTCTCAGTCTCCGGACCTGAACCCGATCGAAAACCTGTGGGCTGAACTGAAAAAACGTGTTCGTGCTCGTCGTCCGACCAACCTGACCCAGCTGCACCAGCTGTGCCAGGAAGAATGGGCTAAAATCCACCCGACCTACTGCGAAAAACTGGTTGAAGGTTACCCGAAACGTCTGACCCAGGTTAAACAGTTCAAAGGTAACGCTACCAAATACTAAG