Part:BBa_K2273113:Design

Short version of the Promoter P_penP found in B. subtilis

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 Notes

This Promoter was designed to be cloned in front of the luxABCDE operon serving a a reporter gene in the pBS3C vector backbone. Therefore, the RFC10 Standard Restriction sites have been added as prefix and suffix via PCR.

RFC10 prefix and suffix sequences:

Prefix with	EcoRI, NotI, XbaI	GAATTCGCGGCCGCTTCTAGA
Suffix with	SpeI, NotI and PstI	ACTAGTAGCGGCCGCTGCAGA

Sites of restriction enzymes generating compatible overhangs are indicated by sharing one color. (EcoRI and PstI are marked in blue, NotI in green, XbaI and SpeI in red

The promoter sequence was amplified from the B.subtilis W168 genome using primers featuring the RFC10 standard restriction sites:

Forward Primer Sequence containing RFC10 prefix: 5`-gatcGGAATTCGCGGCCGCTTCTAGAAATCACAATTGATAAAGCTTTCTAA-3`

Revers Primer Sequence containing RFC10 suffix: 5`-gatcCTGCAGCGGCCGCTACTAGTTCAAATGATTTTGTATTACCTTTG-3`

Source

This part sequence was derived from the Bacillus subtilis W168 genome sequence.

References

[http://www.subtiwiki.uni-goettingen.de/v3/gene/view/713BAB7190E1F86C55103049B29072F00E0DFFB3]

[http://www.uniprot.org/uniprot/P39824]

Marta Toth et al. (2015) Class D Beta-Lactamases Do Exist in Gram-Positive Bacteria. Nature Chemical Biology (12): 9-14.