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Part:BBa_K1442118:Design

Designed by: Iva Burova   Group: iGEM14_Warwick   (2014-10-16)
Revision as of 22:29, 16 October 2014 by Registry (Talk | contribs)

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5' RdRP Promoter


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 292
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 198
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 365


Design Notes

Ribozyme The use of a ribozyme is necessitated due to the complicated binding process between the RNA template and the RdRP. In order to optimise the process and avoid any risk of unfavourable secondary structures or obstruction by unneeded nucleotides, a self-cleaving ribozyme was put after the RNA promoter. The rationale being that after transcription (either in vitro for human cell tests or in vivo for e-coli tests), any added bases as a result of the T7 polymerase would be removed from the main template strand. The chosen ribozyme taken from the Hepatitis Delta virus was investigated at length by the team of J. Doudna and is reported to be the fasted naturally occurring, independent and resistant to denaturants. Its close genetic origin also contribute to a better working and compatible system.



Source

Hepatitis C Virus

References