Difference between revisions of "Part:BBa K2483000"
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This part is an upgrade to the part <a href="https://parts.igem.org/Part:BBa_K515100">BBa_K515100</a> of iGEM Imperial College 2012 producing Indoleacetic acid with the enzymes IaaM (Indoleacetic Acid tryptophan monooxygenase) and IaaH (Indoleacetamide Hydrolase). Here we just fused the IAA enzymes to the RNA-binding proteins (RBPs) MS2 and PP7, respectively. | This part is an upgrade to the part <a href="https://parts.igem.org/Part:BBa_K515100">BBa_K515100</a> of iGEM Imperial College 2012 producing Indoleacetic acid with the enzymes IaaM (Indoleacetic Acid tryptophan monooxygenase) and IaaH (Indoleacetamide Hydrolase). Here we just fused the IAA enzymes to the RNA-binding proteins (RBPs) MS2 and PP7, respectively. | ||
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− | We used this part to produce Indoleacetic Acid (IAA) in E.coli and increase the efficiency of the reaction by putting them in close proximity to each other (metabolic channelling). This was done with a dCas9 (<a | + | We used this part to produce Indoleacetic Acid (IAA) in E.coli and increase the efficiency of the reaction by putting them in close proximity to each other (metabolic channelling). This was done with a dCas9 (<a href="https://parts.igem.org/Part:BBa_K2483002">BBa_K2483002</a>) scaffold where the RNA-binding proteins bind to aptamer sequences added to the 3'-end of a sgRNA and the sgRNA binds to target cassettes where several of the sgRNA recognition sequences are consecutively arranged (<a href="https://parts.igem.org/Part:BBa_K2483005">BBa_K2483005</a> and <a href="https://parts.igem.org/Part:BBa_K2483006">BBa_K2483006</a>). |
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For the complete part with sgRNA, dCas9 and the IAA-enzymes, see https://parts.igem.org/Part:BBa_K2483004. | For the complete part with sgRNA, dCas9 and the IAA-enzymes, see https://parts.igem.org/Part:BBa_K2483004. |
Revision as of 12:47, 1 November 2017
IAA producing enzymes fused to RNA-binding proteins under control of Pveg2
This part is an upgrade to the part BBa_K515100 of iGEM Imperial College 2012 producing Indoleacetic acid with the enzymes IaaM (Indoleacetic Acid tryptophan monooxygenase) and IaaH (Indoleacetamide Hydrolase). Here we just fused the IAA enzymes to the RNA-binding proteins (RBPs) MS2 and PP7, respectively.
We used this part to produce Indoleacetic Acid (IAA) in E.coli and increase the efficiency of the reaction by putting them in close proximity to each other (metabolic channelling). This was done with a dCas9 (BBa_K2483002) scaffold where the RNA-binding proteins bind to aptamer sequences added to the 3'-end of a sgRNA and the sgRNA binds to target cassettes where several of the sgRNA recognition sequences are consecutively arranged (BBa_K2483005 and BBa_K2483006).
For the complete part with sgRNA, dCas9 and the IAA-enzymes, see https://parts.igem.org/Part:BBa_K2483004.
<img src="http://2017.igem.org/File:T--Potsdam--project--abstractskizze.png" width="140px" align="right"/></a>
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 547
Illegal BamHI site found at 1492 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 254
Illegal NgoMIV site found at 3198 - 1000COMPATIBLE WITH RFC[1000]