Template:SBB10 ConstructionFiles DanielS sbb10

PCR DS001 and DS002 on pBca9523-Bca1587 (1051 bp, EcoRI/BamHI) Sub into pBjk2741           (EcoRI/BamHI, 2170+910, L) Product is pBjk2741-sb10         {

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