Part:BBa_K4165173
Biobrick GST - CoH2 - Linker - WWW
This composite part consists of T7 promoter (BBa_K3633015), lac operator (BBa_K4165062), RBS 4 (BBa_K4165264), GST (BBa_K4165070), CoH2 (BBa_K4165003), GGGGSx3 (BBa_K4165068), WWW (BBa_K4165007), and T7 terminator (BBa_K731721).
Source
Synthesized
Usage and Biology
The protac in our snitch system, it is responsible for connecting the target protein (tau) to the tTrim21 which binds to E2 conjugating enzymes and put tau and ubiquitin in proximity to each other resulting in ubiquitination of the tau at lysine residue, after ubiquitination the tau will be degraded by 26S proteosome.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 45
Illegal SapI.rc site found at 189
Dry Lab
Modeling
From 3 models out of 16 Models modeled in 4 software, 1 Model ranked best according to its Q_Mean values 4 and 6. The best Model has values of : score 5 out of 6 , C-Beta deviation = 1 , clash score = 3.08 , molprobitey = 1.35 , Ramachandran favored = 96.31, Ramachandran outlier = 0.79 , Q Mean_4 = -0.01 , Q_Mean_6 = -0.29.
Figure 1. The 3D structure of the COH-Linker-WWW 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
Comparison between the mathematical model of dry lab and the wet lab results of GST COH WWW using sonicator
Comparison between the mathematical model of dry lab and the wet lab results of GST COH WWW using chemical lysis
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