Template:SBB10 ConstructionFiles RaffiH sbb33

Construction of Gentamicin resistance gene (sbb33) BglBrick Part PCR rh03F/rh05R on pLoxGen4  (576 bp, gp = A)  PCR rh04F/rh06R on pLoxGen4   (284 bp, gp = B)  PCR rh03F/rh06R on A+B      (bp 833, EcoRI/BamHI) Sub into pBjk2741-Bca1144  (EcoRI/BamHI, 2170+910, L)  Product is pBjk2741-sbb33   {Promoter.rbs.GenR} rh03F Forward EcoRI for BglBricking GenR  ctctgGAATTCatgAGATCTctagcgcgtcgacataagcc rh04F Removing the BglII site from GenR   cgcgtagtgagatAtatatctatgatc rh05R Removing the BglII site from GenR   gatcatagatataTatctcactacgcg rh06R Reverse BamHI for BglBricking GenR  gcaaaGGATCCctagcgcgtcggccgggaag



JCA Notes

 * correct

Since part isn't genR, replacing with new part: PCR ca1749/rh05R on pG80           (621bp, frag 1) PCR rh04F/ca1750 on pG80           (254bp, frag 2) - PCR ca1749/ca1750 on 1+2           (848bp, Eco/Bam) Sub into pBjk2741 - Product is pBjk2741-sbb44    {Promoter.rbs.genR} - ca1749	Forward cloning of genR from pG80	cagtaGAATTCatgAGATCTCTGGCAGTTCCCTACTCTCG ca1750	Reverse cloning of genR from pG80	ccataGGATCCCTTGAACGAATTGTTAGGTGG rh04F Removing the BglII site from GenR   cgcgtagtgagatAtatatctatgatc rh05R Removing the BglII site from GenR   gatcatagatataTatctcactacgcg