Difference between revisions of "Part:BBa K4061200"
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This is a device prepared for future teams that seek to enhance protein-protein interactions. In this device, we have compiled parts for the modular domains- SH3 binding domain (BBa_K4061110) and the Leucine Zipper domain (BBa_K4061115). Teams must use these modular protein sequences along with their protein CDS (without stop codon) and express these two interacting engineered proteins under a promoter of interest. Here, they are shown to be constitutively expressed. A scaffold protein is required for fortifying the interaction. This scaffold (BBa_K4061015) could be expressed under any desired promoter. In this kit, it is regulated by an inducible promoter- induced by IPTG addition.
 
This is a device prepared for future teams that seek to enhance protein-protein interactions. In this device, we have compiled parts for the modular domains- SH3 binding domain (BBa_K4061110) and the Leucine Zipper domain (BBa_K4061115). Teams must use these modular protein sequences along with their protein CDS (without stop codon) and express these two interacting engineered proteins under a promoter of interest. Here, they are shown to be constitutively expressed. A scaffold protein is required for fortifying the interaction. This scaffold (BBa_K4061015) could be expressed under any desired promoter. In this kit, it is regulated by an inducible promoter- induced by IPTG addition.
  
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<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K4061200 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K4061200 SequenceAndFeatures</partinfo>
  

Revision as of 08:32, 21 October 2021

Scaffold gene kit

This is a device prepared for future teams that seek to enhance protein-protein interactions. In this device, we have compiled parts for the modular domains- SH3 binding domain (BBa_K4061110) and the Leucine Zipper domain (BBa_K4061115). Teams must use these modular protein sequences along with their protein CDS (without stop codon) and express these two interacting engineered proteins under a promoter of interest. Here, they are shown to be constitutively expressed. A scaffold protein is required for fortifying the interaction. This scaffold (BBa_K4061015) could be expressed under any desired promoter. In this kit, it is regulated by an inducible promoter- induced by IPTG addition.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
    Illegal NotI site found at 64
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 214
    Illegal SapI.rc site found at 374


Design Notes

The design considerations for each part involved could be seen at their respective pages. This device compiles these parts. We used constitutive promoter to drive the expression of both engineered proteins (proteins with modified domains) while kept the scaffold under an inducible promoter. Teams however needed to visit each part (protein domain) and attach the domain themselves next to their protein CDS while designing their parts.


Source

Whitaker, W. R. et al. "Engineering Robust Control Of Two-Component System Phosphotransfer Using Modular Scaffolds". Proceedings Of The National Academy Of Sciences, vol 109, no. 44, 2012, pp. 18090-18095. Proceedings Of The National Academy Of Sciences, https://doi.org/10.1073/pnas.1209230109.