Difference between revisions of "Part:BBa K2973002"
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===Usage and Biology=== | ===Usage and Biology=== | ||
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This part was used as a <b>positive control</b> for our <i>in vitro</i> protein synthesis experiments. | This part was used as a <b>positive control</b> for our <i>in vitro</i> protein synthesis experiments. | ||
During our experiments, one of the reporter genes that we tested was EGFP (Enhanced Green Fluorescent Protein). To test its functionality, we added its sequence downstream of Pardee's Toehold Switch 32B. The in vitro transcription/ translation reactions were done using the PURExpress® In Vitro Protein Synthesis kit. To enable translation of the protein from the toehold regulated construct, different amounts of trigger sequence were added in the reaction. Also, a reaction without a trigger sequence was included, as a negative control and a leakage measure. To reduce the cost of the reaction, we lowered the reaction volume from 25 to 7 μL. | During our experiments, one of the reporter genes that we tested was EGFP (Enhanced Green Fluorescent Protein). To test its functionality, we added its sequence downstream of Pardee's Toehold Switch 32B. The in vitro transcription/ translation reactions were done using the PURExpress® In Vitro Protein Synthesis kit. To enable translation of the protein from the toehold regulated construct, different amounts of trigger sequence were added in the reaction. Also, a reaction without a trigger sequence was included, as a negative control and a leakage measure. To reduce the cost of the reaction, we lowered the reaction volume from 25 to 7 μL. | ||
Revision as of 12:40, 19 October 2019
T7-RBS- enhanced green fluorescent protein (EGFP)
This composite part consists of T7 Promoter (BBa_J64997) and T7 Terminator(BBa_K731721), Ribosomal Binding Site(BBa_B0034) and EGFP CDS (BBa_K1094400). EGFP (Enhanced Green Fluorescent Protein) is a version of GFP(green fluorescent protein) that produces enhanced bioluminescence signal in the green zone of the noticeable spectrum and is being used widely as a protein reporter. This protein is produced by the jellyfish (Aequorea Victoria).
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 776
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]