Regulatory

Part:BBa_K2017002

Designed by: Monica Victoria Gutierrez Salazar   Group: iGEM16_Valencia_UPV   (2016-10-03)


35s promoter + 5' region

Promoter of cauliflower mosaic virus (CaMV), used for constitutive expression in plants. It is fused with the 5' region of the N. benthamiana polyubiquitin gene, which has a high expression rate. It can enhance the expression of the coding region.

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 1050
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 1070
  • 1000
    COMPATIBLE WITH RFC[1000]

Usage and Biology

35s is a strong constitutive promoter derived from cauliflower mosaic virus (CaMV). It is widely used in plants to improve the level of the expression of foreign genes effectively in all tissues (1). 5’ region is chosen from the N. benthamiana polyubiquitin sequence (Accession number: Nbv5.1tr6241949). It is used due to its high expression rate in every plant tissue (2). It can help our gene to express itself and ensures that the construction is well expressed. It contains BsmbI and BsaI recognition sites with AATG in 3’ as overhang that allows us to ligate with the amplified target.

Together they work as a promoter that enhances transcription. We ligate it with a target, a linker+luciferase and a terminator to create a gRNA testing system. To check results using this promoter, please see BBa_K2017007.

References

1. Pauli S, Rothnie H, Chen G, He X, Hohn T. The Cauliflower Mosaic Virus 35S Promoter Extends into the Transcribed Region. Journal of Virology. 2004;78(22):12120-12128. 2. Benthamiana Atlas (Internet). Sefapps02.qut.edu.au. 2016 (cited 27 June 2016). Available from: http://sefapps02.qut.edu.au/atlas/tREXXX2new.php?TrID=Nbv5.1tr6241949


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