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The Kozak consensus sequence was originally defined as ACCAUGG following an analysis of the effects of single mutations surrounding the initiation codon (AUG) on translation of the preproinsulin gene (1)
. Subsequent mutagenesis studies and a survey of 699 vertebrate mRNAs extended the consensus sequence for translation initiation to GCCGCCACCAUGG, where the A in the underlined AUG start codon is coordinate 1 and the A at position –3 could also be a G (2)
. Functional studies on preproinsulin and alpha-globin translation in cells indicated that a purine (usually A) in position –3 is crucial for efficient initiation of translation, and in its absence, a G at position +4 is essential (4)
. RNA templates with CCACCAUGG were translated well in a study using the Rabbit Reticulocyte Lysate System (Cat.# L4960) in vitro translation reaction (5)
. In the in vitro translation reaction using the TnT® T7 Quick Coupled Transcription/Translation System (Cat.# L1170, L1171, L5540), PCR products can be used as templates (6)
. The upstream primer for generating the PCR template was designed to contain the T7 promoter for transcription by T7 RNA Polymerase (Cat.# P2075) and the Kozak consensus sequence (CCACCAUG or CCACCAUGG) for efficient translation (6)
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