Part:BBa_K1021016

PT7+crtE

This part is the crtE gene (the first component of the carotenoid biosynthesis pathway) downstream of the T7 promoter. It was designed for use in fungi.

Adapted from KEGG reference pathways

crtE was originally from the 2011 Edinburgh team's biobrick (BBa_K523022) that is a assembly of carotenoid genes crtE, crtB, and crtI, all originating from Pantoea ananatis. crtE encodes geranylgeranyl pyrophosphate synthase, part of the carotenoid biosynthesis pathway, which converts farnesyl pyrophosphate to geranylgeranyl pyrophosphate (Misawa et al., 1990).

The T7 promoter part was originally from the 2007 Ljubljana team (BBa_I712074). It was used in order to assemble multiple genes into a single vector. Fungal promoters are quite long and therefore difficult to assemble, and E. coli will often recombine over and remove direct repeats within DNA sequences. Because the T7 promoter is a short sequence, each of the carotenoid genes could be put behind the T7 promoter and assembled together for maximum simultaneous transcription of the carotenoid genes.

1. Misawa, N., Nakagawa, N., Kobayashi, K., Yamano, S., Nakamura, K., and Harashima, K. 1990. Elucidation of the Erwinia uredovora carotenoid biosynthetic pathway by functional analysis of gene products expressed in Escherichia coli. Journal of Bacteriology 172, 6704-6712. 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
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 776
  
• 1000
  COMPATIBLE WITH RFC[1000]