Difference between revisions of "Part:BBa K422014"
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[[Image:mCyPet.png|thumb|200px|'''Figure 1:''' Fluorescence spectra of mCyPet measured at an excitation wavelength of 425 nm.]] | [[Image:mCyPet.png|thumb|200px|'''Figure 1:''' Fluorescence spectra of mCyPet measured at an excitation wavelength of 425 nm.]] | ||
mCyPet has a distinct fluorescence spectra. Measured at an excitation wavelength of 425 nm with a Fluorescein High Precision Monochromator, maximum emission is detected at 482 nm (Figure 1). | mCyPet has a distinct fluorescence spectra. Measured at an excitation wavelength of 425 nm with a Fluorescein High Precision Monochromator, maximum emission is detected at 482 nm (Figure 1). | ||
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===Cloning strategy=== | ===Cloning strategy=== | ||
Revision as of 10:31, 27 October 2010
mCyPet (AarI A-part)
Biological Background
CyPet is an evolutionary optimized cyan fluorescent protein CFP developed in 2005 by Nguyen and Daugherty. They showed the FRET pair CyPet-YPet to have a 7-fold higher ratiometric FRET signal change than their parental pair.
mCyPet has a mutation in A206K preventing it from forming dimers (Ohashi et al., 20007).
Characterization
mCyPet has a distinct fluorescence spectra. Measured at an excitation wavelength of 425 nm with a Fluorescein High Precision Monochromator, maximum emission is detected at 482 nm (Figure 1).
Cloning strategy
BBF RFC28: A method for combinatorial multi-part assembly based on the Type IIs restriction enzyme AarI. mcyPet-A is an A-part compatible for N-Terminal fusion. For more information see [http://2010.igem.org/Team:ETHZ_Basel/Biology/Cloning].
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal SpeI site found at 733
- 12INCOMPATIBLE WITH RFC[12]Illegal SpeI site found at 733
- 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal SpeI site found at 733
- 25INCOMPATIBLE WITH RFC[25]Illegal SpeI site found at 733
Illegal NgoMIV site found at 697 - 1000COMPATIBLE WITH RFC[1000]