Part:BBa_K1768005:Design
Invertible promoter with RFP reporter
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 115
Illegal NheI site found at 138 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 844
Illegal AgeI site found at 956 - 1000COMPATIBLE WITH RFC[1000]
Design Notes
The inverter works by the Cre/LoxP recombinase pathway isolated from the P1 bacteriophage.
The wild type LoxP site is a 34 base pair recognition element and consists of two 13 bp palindromic sequences which flank an 8bp spacer region. The products of Cre-mediated recombination at loxP sites are dependent upon the location and relative orientation (conferred by the spacer region) of the lox sites. The orientation of the Lox elements determines whether a DNA region is excised or inverted to an antisense orientation. Excision occurs when a DNA region is flanked by the Lox sites that are in the same orientation, while inversion occurs if the elements face opposite directions.
The Lox66 and Lox71 recognition elements are derived from the wild type LoxP but contain mutations in one of each of their palindromic arms. After recombination occurs between these sites a double mutated site is created which is no longer recognised by the Cre enzyme and is thus results in a stable genetic change.
Source
BBa_K1768001 BBa_B0034 BBa_E1010
This part was synthesised by IDT
References
Gilbertson, L. (2003). Cre–lox recombination: Cre-ative tools for plant biotechnology. Trends in biotechnology, 21(12), 550-555.
Ghosh, K., & Van Duyne, G. D. (2002). Cre–loxP biochemistry. Methods, 28(3), 374-383.
Hoess, R. H., & Abremski, K. (1985). Mechanism of strand cleavage and exchange in the Cre-lox site-specific recombination system. Journal of molecular biology, 181(3), 351-362.