Part:BBa_K4165072

T7 promoter-GST-DocS-T7 terminator

This composite part is composed of T7 promoter (BBa_K3633015), lac operator (BBa_K4165062), RBS 4 (BBa_K4165264), GST-tag (BBa_K4165070), DocS (BBa_K3396000), and T7 terminator (BBa_K731721).

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
  COMPATIBLE WITH RFC[12]
• 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
  

Modeling

Model “peptide (GST_DocS) modeled by Rosettafold full_model_407305_3” Ranked the fourth model from our 4 ones with a score 4 out of 6 and values of:

cbeta_deviations	clashscore	molprobity	ramachandran_favored	ramachandran_outliers	Qmean_4	Qmean_6
0	182.78	2.72	98.25	0.7	0.75	1.02

                               Figure 1. The 3D structure of the GST-DOCs model Displayed on Pymol.

Mathematical Modeling

Transcription rate and translation rate under T7 promoter

The mathematical modeling was based on our code for the calculation of transcription and translation according to thermodynamic laws and ODE (You can find it on our wiki) and compared the results with the wet lab.

              Figure 2. 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 (1mM IPTG concentration)

              Figure 3. 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 (2mM IPTG concentration)