Composite

Part:BBa_K3265026:Design

Designed by: Alexander Schanne   Group: iGEM19_UZurich   (2019-10-15)


EutSMN bacterial microcompartment


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1293
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1232
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 1067
    Illegal AgeI site found at 3824
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 3212


Design Notes

This part should be used with a low copy number plasmid backbone

We used this backbone: https://parts.igem.org/Part:BBa_K3265035
It is suitable for protein expression in P. chlororaphis and E.coli

We use the very weak RBS J61100 that seems to work fine with keeping base expression levels below.
Furthermore, we use the more tight araBAD promoter system instead of the LacI which is known to be leaky to get more tight control of the protein expression.

Apart from these changes we kept the part the same as in the original: BBa_K2213012
EutS alone has been shown by Manchester to not be toxic or detrimental for cell growth, we therefore did not change this RBS and left the original one in. What we did is identified the exact location and annotated it in in this part so it is easy to recognize as their part was missing the RBS in the annotation.


Source

IDT, synthesized

References