pCRE-HomeRNApolT7

This vector is a medium copy number, E.coli plasmid, 6547 bp in length.

The plasmid contains:

1) the origin of replicon (pUC ori) responsible for plasmid replication in E. coli cells;

2) the sequence of simian virus (SV40 ori) that allows autonomous (as an episome) replication in mammalian cells expressing the SV40 large T-antigen, such as COS-7 cells;

3) the chimera gene (Km(R)) encoding resistance to kanamicin whose transcription can be initiated both in prokaryotic (Pamp) and mammalian (SV40 early promoter) cells; in mammalian cells its transcription is terminated by HSV TK poly A;

4) the sequence coding for the derivative of bacteriophage T7 RNA polymerase fused with nuclear localization signal (HomeRNApolT7); transcription of this sequence is controlled by the CRE-binding sequence fused to a TATA-like promoter sequence (CRE) and viral transcription terminator signal (SV40 poly A); due to the CRE-binding sequence, production of RNA polymerase derivative can be induced by addition of forskolin into culture media of mammalian cells.

The plasmid is fully compatible with those carrying the p15A replicon.