Part:BBa_K5322003:Design
Constitutive Mfp53 Expression System
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30
Illegal NheI site found at 487
Illegal NotI site found at 451 - 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 137
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
To ensure successful expression of the animal-derived mussel foot protein Mfp53 in BL21(DE3), we conducted codon optimization of the Mfp53 sequence.
Source
The plasmid pET29a-J23119-RBS-Mfp53-T7 employs the pET29a vector, which is commonly used for high-level gene expression in Escherichia coli. The components used in this design, including the J23119 promoter, Mfp5, Mfp3, and the protein linker, are all sourced from the standard biological parts registry of iGEM (BBa_J23119, BBa_K1583002, BBa_K4854000, BBa_K5322001). The mussel foot proteins Mfp5 and Mfp3 are derived from natural marine mussels. These parts were selected for their validated functions and compatibility in synthetic biology applications, facilitating controlled expression within bacterial cells under specific conditions.
References
1. Aich P, An J, Yang B, Ko Y H, Kim J, Murray J, Cha H J, Roh J H, Park K M, Kim K. Chem. Commun., 2018, 54(89): 12642.
2. Zhang W, Yang H, Liu F H, Chen T, Hu G X, Guo D H, Hou Q F, Wu X, Su Y, Wang J B. RSC Adv., 2017, 7(52): 32518.