Part:BBa_K1655000

Propane 1 codes three of the ten enzymes necessary to produce propane in Escherichia coli.

Propane 1 includes 1)acyl-CoA thioester hydrolase (YciA) from Haemophilus influenzae, which catalyzes the reaction: acyl-CoA + H2O <-> CoA + a carboxylate, 2) Carboxylic acid reductase (CAR) from Mycobacterium marinum (strain ATCC BAA-535 / M), which catalyzes the reaction: a carboxylate + reduced acceptor <-> an aldehyde + acceptor + H2O, and 3) maturation factor phosphopantetheinyl transferase (sfp) from Bacillus subtilis, which activates CAR by catalyzing the transfer of a prosthetic phosphopanteine group to CAR.

All coding regions are assembled as an operon after a T7 promoter and and RBS from common cloning vectors (such as pCDF-Duet1 by Novagen). Terminator sequence after the last coding sequence, CAR is also derived from common cloning vectors (e.g. pCDF-Duet1 by Novagen uses this terminator).

All coding sequences have been codon optimized twice for E. coli. First with Thermo Fisher's codon optimizer and then with IDT's codon optimization software.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 2902
  Illegal BamHI site found at 573
  Illegal BamHI site found at 1271
  Illegal BamHI site found at 3791
  Illegal BamHI site found at 3929
  Illegal XhoI site found at 3690
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 1379
  Illegal NgoMIV site found at 3608
  
• 1000
  COMPATIBLE WITH RFC[1000]