Translational_Unit

Part:BBa_K3071022:Design

Designed by: Leung Hei Man   Group: iGEM19_Hong_Kong-CUHK   (2019-10-14)


Recombinant Two-component regulatory system RpfG/RpfC construct with sequencing part


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 349
    Illegal NgoMIV site found at 2647
    Illegal AgeI site found at 1181
    Illegal AgeI site found at 1660
    Illegal AgeI site found at 1744
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 398
    Illegal BsaI.rc site found at 262


Design Notes

Condon optimization for E. coli expression; Correction of the stop codon in the original reported sequence; Two sequencing primers flanked the whole construct for clone screening


Source

-

References

1. Chen, C. H., Lin, N. T., Hsiao, Y. M., Yang, C. Y., & Tseng, Y. H. (2010). Two non-consensus Clp binding sites are involved in upregulation of the gum operon involved in xanthan polysaccharide synthesis in Xanthomonas campestris pv. campestris. Research in microbiology, 161(7), 583-589.

2. Dworkin, J., Jovanovic, G., & Model, P. (1997). Role of upstream activation sequences and integration host factor in transcriptional activation by the constitutively active prokaryotic enhancer-binding protein PspF. Journal of molecular biology, 273(2), 377-388.

3. Rappas, M., Schumacher, J., Beuron, F., Niwa, H., Bordes, P., Wigneshweraraj, S., ... & Zhang, X. (2005). Structural insights into the activity of enhancer-binding proteins. Science, 307(5717), 1972-1975.

4. Ryan, R. P., & Dow, J. M. (2011). Communication with a growing family: diffusible signal factor (DSF) signaling in bacteria. Trends in microbiology, 19(3), 145-152.

5. Transcription Initiation by σ54. (n.d.). Retrieved from http://www.cchem.berkeley.edu/wemmer/research/sigma54.html.

6. Wigneshweraraj, S., Bose, D., Burrows, P. C., Joly, N., Schumacher, J., Rappas, M., ... & Buck, M. (2008). Modus operandi of the bacterial RNA polymerase containing the σ54 promoter‐specificity factor. Molecular microbiology, 68(3), 538-546.

7. Zhou, Y., Asahara, H., Schneider, N., Dranchak, P., Inglese, J., & Chong, S. (2014). Engineering bacterial transcription regulation to create a synthetic in vitro two-hybrid system for protein interaction assays. Journal of the American Chemical Society, 136(40), 14031-14038.

8. Transcription Initiation by σ54. (n.d.). Retrieved from http://www.cchem.berkeley.edu/wemmer/research/sigma54.html.