Coding

Part:BBa_K3505022

Designed by: Venetios Michelioudakis   Group: iGEM20_Thessaly   (2020-10-20)
Revision as of 15:05, 25 October 2020 by Venetios (Talk | contribs) (→‎Design Notes)


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

Level 0 CDS

Fig.1:RraA
Fig.2:The overhangs of this part in the Golden Braid Grammar


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 K3505007. and has overhangs compatible for Golden Braid cloning. The CDS has position B2-B5.

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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