Regulatory

Part:BBa_K2516001

Designed by: Valdemir Kim   Group: iGEM17_NU_Kazakhstan   (2017-10-26)
Revision as of 21:30, 1 November 2017 by Valdemirkim (Talk | contribs)


pHSP70/pRBCS2 combined promoter for C. reinhardtii

Improved part from 2016

This part is an improved version of USP_UNIFESP-Brazil 2016 iGEM team. Link: https://parts.igem.org/Part:BBa_K2136013


The combined constitutive endogenous promoter used for foreign gene expression in C. reinhardtii.

This promoter consists of 5 main parts:

1) HSP70A promoter (1-263) is the first upstream region of Heat Shock Protein 70A from C. reinhardtii. This sequence spacing-dependent and acts as an inhibitor of transcriptional silencing in C. reinhardtii. [1]

2) [Improvement] High-efficiency separator sequence between two promoters (264-272) that provides the best spacing between two promoters, resulting in an increase of transformation rate by 2.6 times relative to original separating sequence. [1]

3) RBCS2 promoter (273-454) is a promoter region of nuclear ribulose bisphosphate carboxylase/oxygenase small subunit gene from C. reinhardtii. It is the most widely utilized promoter for robust transgene expression in C. reinhardtii. [1]

4) 5' untranslated region of the nuclear RBCS2 gene (455-489) from C. reinhardtii acts as an enhancer.

5) [Improvement] The first intron of RBCS2 gene (490-634), it acts as an enhancer and increases expression by 5-6 times [2].

This promoter can be utilized by other iGEM teams as a standardized promoter sequence for constitutive gene expression in C. reinhardtii.

Reference List:

1. Schroda, M., Beck, C. F., & Vallon, O. (2002). Sequence elements within an HSP70 promoter counteract transcriptional transgene silencing in Chlamydomonas. The Plant Journal, 31(4), 445-455.

2. Lumbreras, V., Stevens, D. R., & Purton, S. (1998). Efficient foreign gene expression in Chlamydomonas reinhardtii mediated by an endogenous intron. The Plant Journal, 14(4), 441-447.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 264
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


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