Part:BBa_K4165072
T7 promoter-GST-DocS-T7 terminator
This composite part is composed of T7 promoter (BBa_K3633015), lac operator (BBa_K4165062), RBS (BBa_K4165264), GST-tag (BBa_K4165070), DocS (BBa_K3396000), 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
- 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
Model “peptide (GST_DocS) modeled by Rosettafold full_model_407305_3” Ranked fourth model from our 4 ones with score 4 out of 6 and values of C Beta deviation = 0 , clash score = 182.78 , molprobitey = 2.72 , Ramachandran favored = 98.25 , Ramachandran outlier = 0.7 , Q Mean_4 = 0.75 , Q Mean_6 = 1.02
Figure 1. The 3D structure of GST-DOCs 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 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)
WetaLab Results
Figure 4. Transformed plate of GST Doc + pGS-21a
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