Figure 2

Analysis of EIAV plasmid (A) Blast search revealed for sequences above 2.5 kb the presence of a CAT (Chloramphenicolacetyltransferase). For the longest sequence UN_TR272_len_4326 a second bacterial resistance (AmpR-) conferring a resistance to ß-Lactam antibiotics such as Ampicillin. (B) Plasmid map of the predicted Omniscript RT Kit expression plasmid which was identified as the source of the EIAV pol. Qiagen confirmed that such a plasmid is used for their Omniscript product. The EIAV pol sequence is in-frame with a histidin-tag, flanked by a BamHI and a HindIII restriction site and followed by a lambda t0 terminator. Further downstream a inactive CmR resistance followed by a rrnB T1 Terminator. Further upstream a AmpR promoter together with a ß-lactamase can be found. In front of the Insert is a Ribosomal Binding Site (RBS) with a T5 promoter to ensure strong transcription. The system is induced by a lac operator. The backbone of the plasmid seems to be pDS56/RBSII and therefore the origin of replication may be pBR332. The whole plasmid with the name p6EIAV-RT was created by Dr. Stuart J LeGrice in 1991.