pRK603

pRK603

Description: This TEV protease expression vector is intended to be used for controlled intracellular processing of fusion proteins (TEV protease substrates) in E. coli.

Origin: A pZ derivative (Lutz and Bujard, 1997)

Replicon: p15A

Antibotic Resistance: Kanamycin

Promotor: Synthetic λP_L/tetO

Inducer: anhydrotetracycline (100 µg/liter)

Notes: Expression of the TEV protease gene on pRK603 will be constitutive unless the host strain produces Tet repressor. Regulated expression can be achieved in DH5alphaPro™ or BL21Pro™ cells (Clontech). For best results, perform intracellular processing experiments at 30 °C.

Literature Citations: Kapust, R. B. and Waugh, D. S. (2000). Controlled intracellular processing of fusion proteins by TEV protease. Prot. Expr. Purif. 19: 312-318. Lutz, R. and Bujard, H. (1997). Independent and tight regulation of transcriptional units in Escherichia coli via the LacR/O, the TetR/O and AraC/I1-I2 regulatory elements. Nucl. Acids Res. 25: 1203-10.

Nucleotide Sequence:

Description:	*This TEV protease expression vector is intended to be used for controlled intracellular processing of fusion proteins (TEV protease substrates) in E. coli.*

Origin:	A pZ derivative (Lutz and Bujard, 1997)

Replicon:	p15A

Antibotic Resistance:	Kanamycin

Promotor:	Synthetic λP_L/tetO

Inducer:	anhydrotetracycline (100 µg/liter)

Notes:	Expression of the TEV protease gene on pRK603 will be constitutive unless the host strain produces Tet repressor. Regulated expression can be achieved in DH5alphaPro™ or BL21Pro™ cells (Clontech). For best results, perform intracellular processing experiments at 30 °C.

Literature Citations:	Kapust, R. B. and Waugh, D. S. (2000). Controlled intracellular processing of fusion proteins by TEV protease. Prot. Expr. Purif. 19: 312-318. Lutz, R. and Bujard, H. (1997). Independent and tight regulation of transcriptional units in Escherichia coli via the LacR/O, the TetR/O and AraC/I1-I2 regulatory elements. Nucl. Acids Res. 25: 1203-10.

Nucleotide Sequence: