Translational_Unit

Part:BBa_K3071023:Design

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


Co-activator construct for activating synthetic clp-regulated pspA promoter with sequencing part


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 953
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 956
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 953
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 953
    Illegal NgoMIV site found at 1031
    Illegal AgeI site found at 136
    Illegal AgeI site found at 164
    Illegal AgeI site found at 302
    Illegal AgeI site found at 2509
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Condon optimization for E. coli expression; 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.