Composite
Medium GFP
Part:BBa_K2918032
Designed by: TUDelft2019 Group: iGEM19_TUDelft (2019-10-04)
Medium T7 promoter - Universal RBS - GFP - T7 terminator
A level 1 with Medium T7 promoter, a universal RBS, a GFP and a T7 terminator
Sequence and Features
Assembly Compatibility:
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 465
- 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 465
- 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 465
Illegal BamHI site found at 674 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 465
- 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 465
- 1000COMPATIBLE WITH RFC[1000]
The construct is confirmed by sequencing and there are no mutations.
Toxicity
Our Sci-Phi 29 tool is based on four components of the Φ29 bacteriophage: DNAP, TP, p5 and p6. However, overexpression of these proteins are toxic for the cell. In order to determine the optimal expression levels of the proteins in live cells, we carried out viability assays in E. coli BL21(DE3) pLysS. The results are shown in the graphs below.
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Categories
Parameters
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