Part:BBa_K410000:Design

Periplasmic Heat Generation

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NotI site found at 394
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

Design Notes

Design considerations revolved primarily around targeting the AOX to the periplasm in order to localize AOX near its proper substrates.

Source

The part consists of three sub-parts with varying origins. The cold-shock promoter, HybB, was amplified using PCR from the biobrick BBa_J43001 in the Spring 2010 Distribution. OmpA was amplified from a commercial vector pFLAGS-CTS (Sigma), a plasmid that contains the 81 bp signal peptide. Aox1a, the alternative oxidase from Sacred Lotus, was synthesized commercially by Mr. Gene, Co. The sequence was taken from Genbank (GenBank: AB491175.1) and is from the Sacred Lotus (Nelumbo Nucifera).

References

HybB Sequence [[1]]

OmpA Sequence: http://www.sigmaaldrich.com/etc/medialib/docs/Sigma/Datasheet/e8283dat.Par.0001.File.tmp/e8283dat.pdf

AoxA: Sequence: http://www.ncbi.nlm.nih.gov/nuccore/226897260#sequence_226897260

Papers of Interest:

• Grant, N et al. “Two Cys or Not Two Cys? That is the Question; Alternative Oxidase in the Thermogenic Plant sacred Lotus.” Plant Physiology 150 (2009): 987-995.