Part:BBa_K1351021

Monomeric Red Fluorescent Protein from Discosoma striata

Mutant of the Biobrick BBa_E1010 [[1]]. 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 [http://2012.igem.org/Team:LMU-Munich/Bacillus_BioBricks Bacillus BioBrick Box] by adding different fluorescent proteins in order to have more reporters available in B. subtilis. It still works very well in E. coli, the rather weak coloring you see on the pictures is due to the fact, that a rather homogenous and weak B. subtilis expression vector (pBS0K-Pspac) was used in E. coli

BBa_K1351021 in the B. subtilis expression vector pBS0K-Pspac in E. coli

The left eppendorf tube contains a pink bacterial pellet of E. coli containing the BioBrick BBa_K1351021 in pBS0K-pPspac, a B. subtilis expression vector. On the right, you can see E. coli with the empty vector

Design

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 [http://2014.igem.org/Team:LMU-Munich BaKillus].

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]