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RNA

Part:BBa_K1614016:Design

Designed by: Frieda Anna Sorgenfrei   Group: iGEM15_Heidelberg   (2015-09-16)


Hammerhead Ribozyme and Malchite Green Aptamer in BBF RFC 110 transcription cassette


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 85
    Illegal BamHI site found at 117
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 124
    Illegal NgoMIV site found at 153
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Cloned without restriction enzymes to avoid issues occurring from conserved cut sites. For more details see BBF RFC 110. The part was designed to live track the in vitro transcription of any RNA of interest (ROI). Every ROI can be cloned into this part behind the T7 promotor and in front of the hammerhead ribozyme (HHR) using cloning techniques proposed by our team in BBF RFC 110. This way it is possible so track in real-time how much ROI is transcribed. On top of that it increases the amount of correctly transcribed ROI and results in a defined 3' end as the HHR cleaves off at the defined position. This makes universal measurement device.

Source

Synthetic, Malchit Green aptamer (Grate, D., and Wilson, C. (1999). Laser-mediated, site-specific inactivation of RNA transcripts. Proceedings of the National Academy of Sciences of the United States of America 96, 6131-6136.) HHR (Prody, G.A., Bakos, J.T., Buzayan, J.M., Schneider, I.R., and Bruening, G. (1986). Autolytic processing of dimeric plant virus satellite RNA. Science 231, 1577-1580)

References