Hello Sandeep, the Kozak sequence is the first few nucleotides preceding the start codon (ATG) and the first nucleotide after the start codon (Kozak, 1999). It (A/GCCAUGG) enhances translation from the correct initiation codon. It is to the mammalian cell what the Shine Dalgarno sequence is to prokaryotes. I suggest you include it in your cloning primer.

Hello, Sandeep Kumar,

Kozak consensus sequence ( gcc)gccRccAUGG) is important for eukaryotic translation process (mRNA -> protein) initiation. It is analogous to Shine-Dalgarno sequence in Prokaryotes.

May find this link to other RG discussion useful:

https://www.researchgate.net/post/Is_Kozak_sequence_necessary_for_protein_expression

Hope this help

Martin

Hello Sandeep, the Kozak sequence is the first few nucleotides preceding the start codon (ATG) and the first nucleotide after the start codon (Kozak, 1999). It (A/GCCAUGG) enhances translation from the correct initiation codon. It is to the mammalian cell what the Shine Dalgarno sequence is to prokaryotes. I suggest you include it in your cloning primer.

It require for translation initiation in prokaryotes, so it has to be in your construct.