Part:BBa_K2148017

C.reinhardtii chloroplast rbcL 3UTR w/ Homologies

This part contains the 3'UTR for the rbcL gene found in Chlamydomonas reinhardtii chloroplast genome and has the homology regions right from the trnE2-psbH integenic region with psaA promoter. This is coded as a level-0 3UTR/TERM Phytobrick, which together with other level-0 Phytobrick can form a desired transcriptional unit for transformation, which would also be available for Homologous Recombination.

Usage and Biology

This is a level-0 3UTR/TERM Phytobrick conforming to the Phytobrick Standard. Along with other level-0 Phytobrick, it can be used to create a desired transcriptional unit to transform Chlamydomonas reinhardtii choloroplast.

Caution: In chloroplast rbcL 3'UTR don't serve as transcription terminators as in bacteria, but as elements for transcript maturation/stabilisation. atpB 3'UTR can be used as an alternative. (on advice from Prof. Saul Purton - UCL)

Sequence and Features

Assembly Compatibility:

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

  Illegal NheI site found at 515
  Illegal NheI site found at 1944
  Illegal NheI site found at 2265
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 77
  Illegal BsaI.rc site found at 2670
  

References

Wannathong T, Waterhouse JC, Young REB, Economou CK, Purton S. New tools for chloroplast genetic engineering allow the synthesis of human growth hormone in the green alga Chlamydomonas reinhardtii. Applied Microbiology and Biotechnology. 2016;100:5467-5477. doi:10.1007/s00253-016-7354-6.

Michel Goldschmidt-Clermont, Miche`le Rahire and Jean-David Rochaix. Redundant cis-acting determinants of 3¢ processingand RNA stability in the chloroplast rbcL mRNA ofChlamydomonas. The Plant Journal (2008) 53, 566–577. doi: 10.1111/j.1365-313X.2007.03365.x