DNA

Part:BBa_K1791001

Designed by: Graeme Glaister   Group: iGEM15_Lethbridge   (2015-09-17)


Standard RBS expressing MS2 Coat protein ribozyme affinity purification (RAP)

Standard RBS expressing MS2 Coat protein ribozyme affinity purification (RAP). This part utilizes the expression of C-terminal 6X His tagged fusion MS2 coat protein under a T7 promoter and a standard shine dalgarno sequence for expression of MS2. Downstream there is a T7 promoter followed by a theophylline ribozyme and a MS2 apatamer at the 3' end. Co-expression of the two modules results in MS2 binding to the 3' end of the theophylline ribozyme transcript allowing for the MS2-RNA purification by nickel affinity chromatography. The RNA can be eluted from the complex by the addition of theophylline.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 603
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 95
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
Parameters
None