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Revision as of 12:04, 14 October 2016


Homology-atpA promoter/5UTR/NTAG

A level-0 Phytobrick coded as a promoter/5UTR with a Chlamydomonas reinhardtii chloroplast homology region upstream of the promoter sequence to allow for homologous recombination. The homology region is left from the trnE2-psbH integenic region in the Chlamydomonas reinhardtii chloroplast, which is a region commonly used by experts in the field for high yields of the gene of interest.

Usage and Biology

This part can be used with another level-0 Phytobrick to create a transcriptional unit, which can undergo homologous recombination in Chlamydomonas reinhardtii choloroplast.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 45
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 1339
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 45
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 45
  • 1000
    COMPATIBLE WITH RFC[1000]


Verification

This part was verified in the backbone through a restriction digest which confirmed the desired part size. See gel photograph below, part labelled as HPA1 and the molecular ladder used is a Bioline 1kB ladder.


Result of gel electropheresis

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.