Part:BBa_K4165171

Biobrick GST - CoH2

This composite part consists of T7 promoter (BBa_K3633015), lac operator (BBa_K4165062), pGS-21a RBS (BBa_K4165016), GST tag (BBa_K4165070), CoH2 (BBa_K4165003), and T7 terminator (BBa_K731721).

Source

Synthesized

Usage and Biology

COH protein is a part of the Snitch system, which binds to the GST tag for the purification of the COH.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 1172
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 45
  Illegal SapI.rc site found at 189
  

Dry lab

Modeling

Model “peptide (GST_Coh2) modeled by Alphafold GST_Coh2_f66fb_unrelaxed_rank_3_model_2” Ranked first model from our 4 ones with score 5 out of 6 and values of C Beta deviation = 0 , clash score = 13.59 , molprobitey = 1.64 , Ramachandran favored = 98.31 , Ramachandran outlier = 0.56 , Q Mean_4 = 0.643, Q Mean_6 = 0.914

                             Figure 1. The 3D structure of COH model  Visualized by Pymol.

Mathematical modeling

Transcription rate and translation rate under T7 promotor

the mathematical modeling was based on our code for the calculation of transcription and translation (you can find it in the code section) beside with the estimated results from the wet lab.

                      Figure 1. this figure shows the results from the transcription and translation code showing the 
                        variation of mRNA and protein concentrations with time compared with the wet lab results.
 

WetLab Results

                                          Figure 2. Transformed plate of GST COH