Composite
Part:BBa_K2922031
Designed by: Xingyu Chen Group: iGEM19_XMU-China (2019-10-13)
T7 promoter-RBS(B0034)_kil(Ekil)
Summary
The usage of T7 promoter and RBS is to highly express the Lysis protein for Colicin-E1.
Identification
When we wre building this circuit, we did the nucleic acid gel electrophoresis experiment to verify. After the circuit was built, we sent the plasmid to sequence, and got the correct sequencing. After new molecular cloning experiments, we did Enzyme-Cut identification to certify the plasmid was correct. We used the EcoRI and PstI to cut the plasmid, then we got the target separate fragment-284bp. (Fig.1)
The plasmid was transformed into BL21(DE3).After growing the single colonies,SDS-PAGE protein gel electrophoresis was used to certify the expression of Ekil,then we got the target molecular weight-4.8kDa.(Fig.2)
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 176
- 1000COMPATIBLE WITH RFC[1000]
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Categories
Parameters
None |