Difference between revisions of "Part:BBa K422014"
 
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mCyPet has a mutation in A206K preventing it from forming dimers (Ohashi ''et al.,'' 20007).
 
mCyPet has a mutation in A206K preventing it from forming dimers (Ohashi ''et al.,'' 20007).
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===Characterization===
 
===Characterization===
 
[[Image:mCyPet.png|thumb|200px|'''Figure 1:''' Fluorescence spectra of mCyPet.]]
 
[[Image:mCyPet.png|thumb|200px|'''Figure 1:''' Fluorescence spectra of mCyPet.]]
 
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 shows the relative fluorescene units of mCyPet over a wavelenght range of 460 to 560 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 shows the relative fluorescene units of mCyPet over a wavelenght range of 460 to 560 nm.
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===Cloning strategy===
 
===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].
 
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].
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Latest revision as of 11:14, 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

Figure 1: Fluorescence spectra of mCyPet.

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 shows the relative fluorescene units of mCyPet over a wavelenght range of 460 to 560 nm.

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


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal SpeI site found at 733
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal SpeI site found at 733
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal SpeI site found at 733
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal SpeI site found at 733
    Illegal NgoMIV site found at 697
  • 1000
    COMPATIBLE WITH RFC[1000]