Regulatory
PcomK mut

Part:BBa_K2273013:Design

Designed by: Jonathan Hammer   Group: iGEM17_TU_Dresden   (2017-09-30)


PcomK (mutated): a ComK-dependent promoter


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]


Design

This part was submitted with RFC10 Suffix and Prefix. It encodes for the mutated (mut) comK promoter that is part of the ComX quorum response of B. subtilis and was designed for the subproject "Communication" by the [http://2017.igem.org/Team:TU_Dresden/Project/Communication iGEM Team TU Dresden 2017 (EncaBcillus - It's a trap!)]. In order to remove a SpeI restriction site one nucleotide was exchanged (nt 32, A to T). Therefore it is marked as mutated (mut).

Source

The comK promoter was amplified via PCR from the B. subtilis wild type W168 genome using the primers listed below.

PcomK mut fwd RFC10 gatcTCTAGAGCAAGTCTTATGAAAGTAAAATCGGTTTATTTCTAG

This primer only contains the XbaI restriction site of the prefix. The nucleotide marked bold was exchanged (A to T) in order to remove a SpeI restriction site.

PcomK mut rev RFC10 gatcCTGCAGCGGCCGCTACTAGTTTTTCTGCAAAATTTATACTAATAATC

References

Susanna KA, van der Werff AF, den Hengst CD, Calles B, Salas M, Venema G, Hamoen LW, Kuipers OP.(2014) Mechanism of transcription activation at the comG promoter by the competence transcription factor ComK of Bacillus subtilis. J Bacteriol. 186(4):1120-8 PubMed

Hamoen LW, Venema G, Kuipers OP (2003) Controlling competence in Bacillus subtilis: shared use of regulators. Microbiology. 149(Pt 1):9-17 PubMed