The pET-46 Ek/LIC vector is prepared for rapid, directional cloning of PCR-amplified DNA for highlevel expression of polypeptides. Using specifically designed primers for amplification and the pET-46 Ek/LIC Cloning Kit (Cat. No. 71335-3), inserts can be efficiently cloned without the need for restriction digestion or ligation. Fusion proteins contain an N-terminal cleavable His?Tag® sequence. Fusion to an optional C-terminal S?TagTM sequence can also be created for detection, purification and quantification of fusion proteins. Unique sites are shown on the circle map. Note that the sequence is numbered by the pBR322 convention, so the T7 expression region is reversed on the circle map. The f1 origin is oriented so that infection with helper phage will produce virions containing single stranded DNA that corresponds to the coding strand. Therefore, single stranded sequencing should be performed using the T7 Terminator Primer (Cat. No. 69337-3).