Difference between revisions of "Part:BBa K1645999"

Revision as of 00:14, 20 October 2016

Dual Color Plasmid (RFP and GFP)

This part is a composite of an RFP gene and a GFP gene from the previously made BioBricks: BBa_I20260 and BBa_J04450, respectively.

It can be used in CRISPRi experiments to demonstrate the effectiveness of sgRNA-Cas9 targeting. It can also be used for other gene expression suppression systems to provide quantitative feedback and data.

It has been characterized using flow cytometry to demonstrate the expression of both RFP and GFP proteins.

Further characterization of this part being used in a CRISPRi system can be found with the BBa_K1645998 BioBrick.

Characterization

File:Https://static.igem.org/mediawiki/parts/0/06/T--Waterloo iGEM--322-RFP-GFP 2016.jpeg

Figure 1: First characterization experiment of the KlADH4-promoter (& eGFP) in S. cerevisiae. After an overnight pre culture, the transformed yeast cells were transferred into SC-U Media with different ethanol concentration and the eGFP-fluorescence and the OD600 were measured at different times.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 7
Illegal NheI site found at 30
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal AgeI site found at 1708
Illegal AgeI site found at 1820
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI.rc site found at 706

@@ Line 12: / Line 12: @@
 ==Characterization==
-[[Image:T--Waterloo_iGEM--322-RFP-GFP_2016.jpg|400px|thumb|right|Figure 1: First characterization experiment of the KlADH4-promoter (& eGFP) in ''S. cerevisiae''. After an overnight pre culture, the transformed yeast cells were transferred into SC-U Media with different ethanol concentration and the eGFP-fluorescence and the OD600 were measured at different times.]]
+[[Image:https://static.igem.org/mediawiki/parts/0/06/T--Waterloo_iGEM--322-RFP-GFP_2016.jpeg|400px|thumb|right|Figure 1: First characterization experiment of the KlADH4-promoter (& eGFP) in ''S. cerevisiae''. After an overnight pre culture, the transformed yeast cells were transferred into SC-U Media with different ethanol concentration and the eGFP-fluorescence and the OD600 were measured at different times.]]
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