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pNLCoI3[luc2-P2A-NlucP/CMV/Hygro] Vector

Instructions for Use of Product(s)

Literature # 9PIN148

The pNLCoI3[luc2-P2A-NlucP/CMV/Hygro] Vector is a second generation coincidence reporter vector (CoI) that allows expression of both firefly luciferase (Fluc, luc2 gene) from Photinus pyralis and NanoLuc® Luciferase fused to a PEST destabilization domain (NanoLuc®-PEST, NlucP gene ) from the same mRNA transcript. The stoichiometric expression of both luciferases is achieved by use of the P2A sequence from porcine teschovirus-1, which promotes a ribosomal skip and expression of the two unfused enzymes with different compound interaction profiles. When used in highthroughput screening, true hits will show a similar response from both reporters, helping to reduce the number of false hits and the workload associated with follow-up screens.

The luc2 and NlucP genes are codon optimized for mammalian expression, and all pNL vectors have minimal consensus transcription factor-binding sites to reduce anomalous expression. The pNLCoI3[luc2-P2A-NlucP/CMV/Hygro] Vector contains a CMV promoter for high-level expression of both reporters, allowing its use as a constitutive expression control.

Printed in USA. Revised 9/16