The pET-38b(+) vector (Cat. No. 70137-3) is designed for expression of C-terminal CBD cex fusion proteins. Target sequences inserted between the N-terminal signal sequence and C-terminal CBD cex •Tag™ and His•Tag ® sequences allow for potential export of the expressed protein. Sequences inserted at the Nde I site will be expressed in the cytoplasm. Note that inserts must allow read-through in the proper reading frame (shown below) to produce the desired polypeptide fusions. A variety of cloning sites and strategies are available. 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 cloning/expression region of the coding strand transcribed by T7 RNA polymerase is shown below. 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).
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