Vector Maps and Sequences

Basic Science Research


Vector Tool box:

Vector maps and sequences are available for free download as PDF files or Vector NTI files by clicking on the relevant link below for each vector.

Description Vector Name NTI File PDF
Dual promoter reporter vector Lenti pHAGE CMV-dsRed-UBC-GFP-W Download map/sequence as an NTI file Download map/sequence as a PDF file
Dual promoter alpha-1 antitrypsin (AAT-GFP) Lenti pHAGE CMV-AAT-UBC-GFP-W Download map/sequence as an NTI file Download map/sequence as a PDF file
AAT-ZsGreen Lenti pHAGE UBC-AAT-IRES-ZsGreen-W Download map/sequence as an NTI file Download map/sequence as a PDF file
Single Promoter AAT Lenti pHAGE CMV-AAT-W Download map/sequence as an NTI file Download map/sequence as a PDF file

The lentiviral system we employ is based on an HIV-1-based backbone, named ‘pHAGE’ (standing for plasmid HIV-1 Alex Gustavo George Enhanced) originally developed in the laboratory of Dr. Richard C. Mulligan at Harvard Medical School. Dr. Kotton worked with the original pHAGE backbone during his postdoctoral fellowship in the Mulligan laboratory and the backbone has since been adapted to express the transgenes indicated on this page.

We have previously published a detailed description of the lentiviral backbone and technique for packaging high titer virus for in vivo studies, and several of these protocols may be downloaded here by clicking on protocols.

To summarize, the third generation lentiviral vectors employed in our proposal are typically pseudotyped with a VSV-G envelope that allows stability and tropism for cells of multiple species including mice. The engineered virus is replication incompetent and has a self-inactivating viral promoter (deleted U3). This ensures that, after integration in the target cell genome, only the internal promoter of interest is active and no further viral replication or infection occurs. A woodchuck hepatitis virus post-transcriptional regulatory element (WPRE) and cPPT is included in each vector to increase gene expression levels in transduced cells.