Difference between revisions of "Part:BBa K3229303"
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This part is constructed to certify the bidirectional transcription from McyD promoter. Normally microcystin toxin-producing genes (McyABC and McyDEFGHIJ) are located next to the McyD promoter in both directions. When microcystin toxin is present, it induces the synthesis of the composite GFP parts due to the phenomenon called autoinduction. We replaced the toxin-producing genes with a composite GFP, which consists of two parts. We put one part in the 3' direction and the other one in the 5' direction from the McyD promoter. When the two parts are synthesized at the same time they connect together to form an active GFP that glows. Thus The transformed bacteria that has the insert can only glow under UV light if the bidirectional transcription works. This construct consists of the McyD promoter, an RBS, GFP1 and a terminator sequence in the 3' direction from the promoter and a reverse RBS, a reverse GFP2 and a reverse terminator sequence in the 5' direction from the promoter. Our intention is to transform Microcystis aeruginosa beside E. coli. A shuttle plasmid that can replicate in both species is required for this procedure. In order to insert this construct into the shuttle plasmid we put a non-coding ATATATATAT sequence and a BamHI restriction site at the end of our construct. | This part is constructed to certify the bidirectional transcription from McyD promoter. Normally microcystin toxin-producing genes (McyABC and McyDEFGHIJ) are located next to the McyD promoter in both directions. When microcystin toxin is present, it induces the synthesis of the composite GFP parts due to the phenomenon called autoinduction. We replaced the toxin-producing genes with a composite GFP, which consists of two parts. We put one part in the 3' direction and the other one in the 5' direction from the McyD promoter. When the two parts are synthesized at the same time they connect together to form an active GFP that glows. Thus The transformed bacteria that has the insert can only glow under UV light if the bidirectional transcription works. This construct consists of the McyD promoter, an RBS, GFP1 and a terminator sequence in the 3' direction from the promoter and a reverse RBS, a reverse GFP2 and a reverse terminator sequence in the 5' direction from the promoter. Our intention is to transform Microcystis aeruginosa beside E. coli. A shuttle plasmid that can replicate in both species is required for this procedure. In order to insert this construct into the shuttle plasmid we put a non-coding ATATATATAT sequence and a BamHI restriction site at the end of our construct. | ||
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Revision as of 16:29, 21 October 2019
McyA/D bidirectional promoter 1
This part is constructed to certify the bidirectional transcription from McyD promoter. Normally microcystin toxin-producing genes (McyABC and McyDEFGHIJ) are located next to the McyD promoter in both directions. When microcystin toxin is present, it induces the synthesis of the composite GFP parts due to the phenomenon called autoinduction. We replaced the toxin-producing genes with a composite GFP, which consists of two parts. We put one part in the 3' direction and the other one in the 5' direction from the McyD promoter. When the two parts are synthesized at the same time they connect together to form an active GFP that glows. Thus The transformed bacteria that has the insert can only glow under UV light if the bidirectional transcription works. This construct consists of the McyD promoter, an RBS, GFP1 and a terminator sequence in the 3' direction from the promoter and a reverse RBS, a reverse GFP2 and a reverse terminator sequence in the 5' direction from the promoter. Our intention is to transform Microcystis aeruginosa beside E. coli. A shuttle plasmid that can replicate in both species is required for this procedure. In order to insert this construct into the shuttle plasmid we put a non-coding ATATATATAT sequence and a BamHI restriction site at the end of our construct.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 883
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 113