Part:BBa_K2516001
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
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 264
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
| None |