pPhsp70-HomeRNApolT7-12

This Escherichia coli mammalian cell shuttle vector enables expression of genes controlled by bacteriophage T7 promoter in mammalian cells.

This vector is a medium copy number, E.coli plasmid, 7598 bp in length.

The plasmid contains:

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

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 human heat shock promoter (E1 and E2) and SV40 transcription terminator (SV40 poly A).

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