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) (3) . 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) .