Improved Parts

BBa_K2443037 - T7 RNA Polymerase

Original part: BBa_I2032

Submitted by: MIT in 2006

designed by: Bartholomew Canton

Rational behind improvements:

Encodes only for the coding region of T7 RNA polymerase. In order to improve it and incorporate it into our NextVivo system we have codon optimized it and attached a 6 X His tag for easy purification.

Part BBa_K1791002

Part BBa_K17910012 contains two modules, a low efficiency ribosome binding site (RBS) expressing MS2 coat protein and a theophylline ribozyme with a MS2 aptamer at the 3’ end. To express MS2 coat protein the part utilizes expression of c-terminal 6x His tagged MS2 coat protein under a T7 promoter and a standard Shine Dalgarno sequence followed by a double terminator (BBa_0015). Downstream of the RBS is a T7 promoter followed by a theophylline ribozyme and a MS2 aptamer at the 3’ end. Coexpression of the two modules results in MS2 binding to the 3’ end of the theophylline ribozyme construct which allows for MS2-RNA purification by nickel affinity chromatography, RNA can then be eluted from the MS2-RNA complex by addition of theophylline.