SIL072006-Construction of pSB1A2-J23015
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F plasmid with OriT knockout
PCR FORlamF/ForlamR on pKD3 (~1000bp, dPN1, Neb2)
Electroporate into pKD46xpOX38
Product is pSB1A2-J23015
ForlamF Forward oligo to knockout OriT:
tttatccgtaaataatttaacccactccacaaaaaggctcaacag gtgtaggctggagctgcttc
ForlamR Reverse oligo to knockout OriT:
ggaccatggctaattcccat ctctttatgatatttaaaaaagcggtgtcggcgcggctacaacaa