Coding

Part:BBa_K3505022

Designed by: Venetios Michelioudakis   Group: iGEM20_Thessaly   (2020-10-20)
Revision as of 20:49, 27 October 2020 by Asteriat (Talk | contribs) (→‎Design Notes)


RraA- Regulator of ribonuclease activity A GB compatible with B2-B5

Level 0 CDS

Fig.1:RraA


Usage and Biology

The Regulator of ribonuclease activity A (RraA), a repressor of the mRNA-degrading ability of the E. coli RNase E. [1] This protein leads to better expression of non omologous membrane proteins in E. coli.

Design Notes

The coding sequence was domesticated . We removed BsmBI ,BsaI , BtgZI, BpiI sites in order to be compatible with GoldenBraid and MoClo. The sequence is cloned in pUPD2 BBa_K3505007 and has overhangs compatible for GoldenBraid cloning. The CDS has position B2-B5.

Fig.2:The overhangs of this part in the GoldenBraid Grammar

Verification of cloning

Fig.4:Level 0 RraA U2 C2 Digested with EcoRI, PstI Expected bands 2029, 564

Experimental Use and Experience

This part is used in BBa_K3505038

=Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]

References

  • [1]Gialama, D., Delivoria, D., Michou, M., Giannakopoulou, A. and Skretas, G., 2017. Functional Requirements for DjlA- and RraA-Mediated Enhancement of Recombinant Membrane Protein Production in the Engineered Escherichia coli Strains SuptoxD and SuptoxR. Journal of Molecular Biology, 429(12), pp.1800-1816.
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