Template:SBB10 ConstructionFiles RaffiH sbb33

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 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)
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 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}
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 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
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