Coding

Part:BBa_K3505021

Designed by: Venetios Michelioudakis   Group: iGEM20_Thessaly   (2020-10-20)


DjlA- DnaJ-like protein A GB compatible with B2-B5

Level 0 CDS

Fig.1:DjlA


Usage and Biology

DnaJ-like protein A (DjiA) is a membrane-bound DnaK co-chaperone that has antitoxic capacities for big protein expression in bacteria.[1]


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 Golden Braid cloning. The CDS has position B2-B5.

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

Verification of cloning

Fig.3:Level 0 DjlA U4 C4 Digested with EcoRI, PstI Expected bands 2029, 894

Experimental Use and Experience

This part is used in BBa_K3505039

Source

Georgios Skretas [1]

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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Parameters
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