![]() ![]() ( C) the cytomegalovirus (CMV) early enhancer element, ( A) the promoter, the first exon and the first intron of chicken beta-actin gene, ( G) the splice acceptor of the rabbit beta-globin gene ĬAG promoter was constructed in the lab of Dr Jun-ichi Miyazaki from the following sequences: The 1-base overhangs produced by BmrI may be hard to ligate.The CAG promoter is a strong synthetic promoter frequently used to drive high levels of gene expression in mammalian expression vectors. Sticky ends from different BglI sites may not be compatible.Ī C T G G G ( N ) 4 N T G A C C C ( N ) 4 G C C N N N N N G G C C G G N N N N N C C G Sticky ends from different BstAPI sites may not be compatible.īsaHI is typically used at 37☌, but is even more active at 60☌.Įfficient cleavage requires at least two copies of the NarI recognition sequence.Įfficient cleavage requires at least two copies of the PluTI recognition sequence. G C A N N N N N T G C C G T N N N N N A C G Prolonged incubation with NdeI may lead to removal of additional nucleotides. Sticky ends from different PfoI sites may not be compatible. ![]()
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