Vectors

pEGAD Binary expression vector for creating C-terminal fusions to GFP.
BASTA plant selection.
12.5 KB, low copy origin of replication.
pEGAD was created for directional cloning of cDNA libraries at the C-terminus of GFP using the EcoR1 and HindII sites (see Cutler et al, 2000). Unique sites also flank the promoter for easy replacment of sequences upstream of GFP. A polyalanine sequence at the C-terminus of GFP provides a flexible linker arm between GFP and the added peptide sequence.
pEGAD is now available at the stock center.
Binary expression vectors for creating N or C-terminal fusions to GFP.
BASTA plant selection.
9.5 KB, intermediate copy origin of replication.
pEZT
The pEZT vectors are designed for the construction and in planta expression of translational fusions to either the N or C terminus of EGFP. In the "L" series, a polyalanine linker sequence is inserted between GFP and the MCS to create a flexible arm between GFP and the cloned gene. Both vectors have also been made without the added polyalanine sequence. Three identical cloning sites are provided upstream or downstream of EGFP to simplify the design of cloning strategies and primers for making both N and C terminal fusions. Expression is driven from the califlower mosaic virus 35S promoter. The expression cassette is contained within T-DNA ends, and in planta selection for the presence of the T-DNA sequence is conferred by Basta resistence. The plasmid backbone is derived from pBBR, a medium copy plasmid isolated from Bordatella pertussis that has greater stability than conventional braod host range plasmids based on the pRK2 origin of replication. The "NL" vector does not express GFP well without adding coding seqeunce to the 5' end of the open reading frame.
Vectors for creating N or C-terminal fusions to GFP on Not1 cassettes
Good for transient expression experiments
5.7 KB, high copy origin of replication
pEZS
The pEZS vectors consist of the same expession cassettes present in the "T" series, without the plant selection nd T-DNA ends. These plasmids are smaller and are higher copy, making them good tools for producing DNA for transient expression studies. The expression cassettes are flanked by Not1 sites for easy subcloning into other plasmid backgrounds. The "NL" vector does not express GFP well without adding coding seqeunce to the 5' end of the open reading frame.

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