Part:BBa_K2100021
pENTR EGSH
This entry vector contains the EGSH promoter. pEGSH is part of the ecdysone inducible expression system. The synthetic receptor fusion VgEcR forms a complex with RXR which binds as a heterodimer to the E/GRE recognition sequences on the EGSH promoter, transcribing the gene of choice downstream of the promoter. This system can be induced with the small molecule PonA, and is otherwise tightly controlled by corepressors.[1]
We constructed a pEGSH vector so we could use it to test how an inducible promoter can modulate the activity of recombinases, ideally keeping basal expression to a minimum. pEGSH was selected as our inducible promoter of choice because it is a steroid-inducible promoter which, according to the literature, exhibits greater control over downstream expression in comparison to other common inducible promoters such as TET. As a steroid-inducible promoter, pEGSH is an ideal analog to pERE, the estrogen responsive promoter that will be used in our final circuit. When consulting with members and advisors of the BU Wetlab 2016 iGEM team, we were told that the inducible promoter TET had shown little success when used in combination with recombinases because of high basal expression.
[1] https://www.agilent.com/cs/library/usermanuals/Public/217564.pdf
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 6
Illegal EcoRI site found at 671
Illegal PstI site found at 485 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 6
Illegal EcoRI site found at 671
Illegal PstI site found at 485 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 6
Illegal EcoRI site found at 671
Illegal BglII site found at 115
Illegal BamHI site found at 663 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 6
Illegal EcoRI site found at 671
Illegal PstI site found at 485 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 6
Illegal EcoRI site found at 671
Illegal PstI site found at 485 - 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 646
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