Difference between revisions of "Part:BBa J100205"

Revision as of 19:17, 2 August 2017

repClone Red

We designed repClone Red to test the function of repressors. Internal BsaI sites allow the insertion of the Ptet promoter and removal of the backwards P2 promoter pointed towards GFP. Successful cloning of the Ptet promoter will change colonies from green to not green. Variations on the Ptet promoter, or introduction of anhydrotetrocycline (aTc), will affect the activity of the repressor, as indicated by the fluorescence of colonies containing the plasmid. This can be used in synthetic biology laboratories or for undergraduate introductory biology lab modules.
We found that JM109 spontaneously deleted the transcriptional terminator (TT). This part was more stable, the cells grew better and mini preps yielded more plasmid. Therefore we recommend people use J100324 instead.

Variations from the registry sequence for TetR and GFP were sequence verified, and functionality was experimentally verified.

Sequence and Features

Assembly Compatibility:

10
INCOMPATIBLE WITH RFC[10]
Illegal SpeI site found at 9
12
INCOMPATIBLE WITH RFC[12]
Illegal SpeI site found at 9
21
COMPATIBLE WITH RFC[21]
23
INCOMPATIBLE WITH RFC[23]
Illegal SpeI site found at 9
25
INCOMPATIBLE WITH RFC[25]
Illegal SpeI site found at 9
Illegal AgeI site found at 2288
Illegal AgeI site found at 2400
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI site found at 1638
Illegal BsaI.rc site found at 1517

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 We designed repClone Red to test the function of repressors. Internal BsaI sites allow the insertion of the Ptet promoter and removal of the backwards P2 promoter pointed towards GFP. Successful cloning of the Ptet promoter will change colonies from green to not green. Variations on the Ptet promoter, or introduction of anhydrotetrocycline (aTc), will affect the activity of the repressor, as indicated by the fluorescence of colonies containing the plasmid. This can be used in synthetic biology laboratories or for undergraduate introductory biology lab modules. <br>
-We found that JM109 spontaneously deleted the transcriptional terminator (TT). This part was more stable, the cells grew better and mini preps yielded more plasmid. Therefore we recommend people use J100324 instead.
+We found that JM109 spontaneously deleted the transcriptional terminator (TT). This part was more stable, the cells grew better and mini preps yielded more plasmid. Therefore we recommend people use [https://parts.igem.org/Part:BBa_J100324 J100324] instead.
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