Monomeric Red Fluorescent Protein from Discosoma striata
Mutant of the Biobrick BBa_E1010. In order to make it compatible with the Freiburg-Standard RFC25, two AgeI-Restriction sites were deleted.
monomeric RFP: Red Fluorescent Protein.
Excitation peak: 584 nm Emission peak: 607 nm
Usage and Biology
This part was created in order to enhance the Bacillus BioBrick Box by adding different fluorescent proteins to have more reporters available in B. subtilis.
Since the Biobrick BBa_E1010 is already evaluated very well and shows a very bright color in E. coli, the rather light coloring in our experimental setup may be due to the used expression vector pBS0K-Pspac. This vector is optimized for the use in B. subtilis and shows a rather weak and heterogenous expression in E. coli.
Picture 1: BBa_K1351021 in the B. subtilis expression vector pBS0K-Pspac, cloned into E. coli. The rather weak color is due to the vector, which shows a rather weak an heterogenous expression. On the right picture, the left eppendorf tube contains a sample of E. coli containing the Biobrick BBa_K1351021, the right eppendorf tube contains E. coli with just the empty vector.
This part was generated in a modified version of RFC25, where a strong Shine Dalgarno Sequence (SD) is included, and has the following prefix and suffix:
|prefix with EcoRI, NotI, XbaI, SD and NgoMIV:||GAATTCGCGGCCGCTTCTAGAGTAAGGAGGAGCCGGC|
|suffix with AgeI, SpeI, NotI and PstI:||ACCGGTTAATACTAGTAGCGGCCGCTGCAG|
Sites of restriction enzymes generating compatible overhangs have the same color:
EcoRI and PstI in blue, NotI in green, XbaI and SpeI in red, NgoMIV and AgeI in orange. Shine-Dalgarno sequence and stop codons are underlined.
This part is used in the 2014 LMU-Munich iGEM project BaKillus.
|Protein data table for BioBrick BBa_ automatically created by the BioBrick-AutoAnnotator version 1.0|
|Nucleotide sequence in RFC 25, so ATGGCCGGC and ACCGGT were added (in italics) to the 5' and 3' ends: (underlined part encodes the protein)|
ATGGCCGGCATGGCTTCC ... ACCTGTGCTACCGGT
ORF from nucleotide position -8 to 681 (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)|
There were no alignments for this protein in the data base. The BLAST search was initialized and should be ready in a few hours.
|Predictions (obtained from PredictProtein.org)|
|There were no predictions for this protein in the data base. The prediction was initialized and should be ready in a few hours.|
| The BioBrick-AutoAnnotator was created by TU-Munich 2013 iGEM team. For more information please see the documentation.|
If you have any questions, comments or suggestions, please leave us a comment.
Sequence and Features
- 10COMPATIBLE WITH RFC
- 12COMPATIBLE WITH RFC
- 21COMPATIBLE WITH RFC
- 23COMPATIBLE WITH RFC
- 25COMPATIBLE WITH RFC
- 1000COMPATIBLE WITH RFC