The pET-28a-c(+) vectors carry an N-terminal His•Tag®/thrombin/T7•Tag® configuration plus an optional C-terminal His•Tag sequence. 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 circular 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).
pET-28a(+) sequence landmarks T7 promoter 370-386 T7 transcription start 369 His Tag coding sequence 270-287 T7•Tag coding sequence 207-239 Multiple cloning sites (BamH I – Xho I) 158-203
His Tag coding sequence 140-157 T7 terminator 26-72 lacI coding sequence 773-1852 pBR322 origin 3286 Kan coding sequence 3995-4807 f1 origin 4903-5358 The maps for pET-28b(+) and pET-28c(+) are the same as pET-28a(+) (shown) with the following exceptions:
pET-28b(+) is a 5368bp plasmid; subtract 1bp from each site beyond BamH I at 198. pET-28c(+) is a 5367bp plasmid; subtract 2bp from each site beyond BamH I at 198.