green fluorescent protein derived from jellyfish Aequeora victoria wild-type GFP (SwissProt: P42212
GFP (mut3b) [note that this part does not have a barcode]
Usage and Biology
The original citation for GFPmut3b is as follows:
Cormack, B.P., Valdivia, R.H., and S. Falkow. FACS-optimized mutants of green fluorescent protein (GFP). Gene 173: 33-38 (1996).
Here's the link: http://www.sciencedirect.com/science/article/pii/0378111995006850
Cormack et al. report the following excitation and emission data for GFPmut3 -
- Excitation max - 501nm
- Emission max - 511nm
Cormack et al. report detectable fluorescence within 8 mins. Please add maturation time data for E0040 here.
Part Characteristics in Cell-Free Chassis
|Parameter||Value and Description|
|Calibration||A conversion factor of 79.429 from Au to concentraion in nM|
|Half-life||33 hours in the cell-free chassis, with a degradation constant of 0.0210 (in hours)|
The fluorescence of purified GFPmut3B was calibrated in the cell-free chassis. The derived calilbration curveallows the determination of the concentration of GFPmut3b in the cell-free chassis. Detailed protocolsfor generating the calibration curve are available. Other calibration curves for are also available on the results page.
The degradation of GFPmut3B in the cell-free chassis was also characterized. Purified GFPmut3B was allowed to degrade in the cell-free chassis and the fluorescence was measured over time. Detailed protocolsand resultsare attached.
From the semi-log plot, the degradation constant (in minutes) was derived to be 0.0003501, which is equivalent to GFPmut3b having a half-life of 33 hours in the cell-free chassis.
|Protein data table for BioBrick BBa_E0040 automatically created by the BioBrick-AutoAnnotator version 1.0|
|Nucleotide sequence in RFC 10: (underlined part encodes the protein)|
ATGCGTAAA ... CTATACAAATAATAA
ORF from nucleotide position 1 to 714 (excluding stop-codon)
|Amino acid sequence: (RFC 25 scars in shown in bold, other sequence features underlined; both given below)|
|Sequence features: (with their position in the amino acid sequence, see the list of supported features)|
|Amino acid composition:|
|Amino acid counting|
|Plot for hydrophobicity, charge, predicted secondary structure, solvent accessability, transmembrane helices and disulfid bridges|
|Alignments (obtained from PredictProtein.org)|
|Predictions (obtained from PredictProtein.org)|
|Subcellular Localization (reliability in brackets)|
|Gene Ontology (reliability in brackets)|
| The BioBrick-AutoAnnotator was created by TU-Munich 2013 iGEM team. For more information please see the documentation.|
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