Difference between revisions of "Part:BBa K2235003"
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<partinfo>BBa_K2235003 short</partinfo> | <partinfo>BBa_K2235003 short</partinfo> | ||
| + | This composite part encodes all the necessary regulatory elements for a functional chemically inducible system, the cumate system. In particular, there is an expression cassette encoding the reporter gene, mTagBFP, <partinfo>K592100</partinfo> and there are two additional multiple cloning sites (MCS) flanking the reporter (check the design page for more information). The repressor of the system, CymR <partinfo>K415202</partinfo> , is located upstream of the expression cassette and it is constitutively expressed. The RNA spinach, <partinfo>K1330000</partinfo>, is an additional reporter protein following CymR aiming to detect its expression upon addition of its cognate fluorophore. | ||
===Usage and Biology=== | ===Usage and Biology=== | ||
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| + | The cumate system have been previously described as a versatile and tightly regulated expression system induced by 4-isopropylbenzoic acid (cumate) in various strains of E.coli (1). It can be employed for the regulation and production of heterologous proteins in a dose-dependent manner. Moreover, the induction is achieved quickly within minutes therefore, a robust and homogenous expression of any protein can be achieved upon addition of cumate. | ||
Latest revision as of 12:53, 23 October 2017
Cumate gene switch expressing BFP
This composite part encodes all the necessary regulatory elements for a functional chemically inducible system, the cumate system. In particular, there is an expression cassette encoding the reporter gene, mTagBFP, BBa_K592100 and there are two additional multiple cloning sites (MCS) flanking the reporter (check the design page for more information). The repressor of the system, CymR BBa_K415202 , is located upstream of the expression cassette and it is constitutively expressed. The RNA spinach, BBa_K1330000, is an additional reporter protein following CymR aiming to detect its expression upon addition of its cognate fluorophore.
Usage and Biology
The cumate system have been previously described as a versatile and tightly regulated expression system induced by 4-isopropylbenzoic acid (cumate) in various strains of E.coli (1). It can be employed for the regulation and production of heterologous proteins in a dose-dependent manner. Moreover, the induction is achieved quickly within minutes therefore, a robust and homogenous expression of any protein can be achieved upon addition of cumate.
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 418
Illegal XhoI site found at 199 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 101
References
1. Choi Y, Morel L, Le Francois T, Bourque D, Bourget L, Groleau D et al. Novel, Versatile, and Tightly Regulated Expression System for Escherichia coli Strains. Applied and Environmental Microbiology. 2010;76(15):5058-5066.