Part:BBa_K2306011:Design
Cytosolic-abundant heat soluble protein 94205 "CAHS 94205" (T7 promoter, RBS and double terminator)
This composite BioBrick features the basic part BBa_k2306006 that codes for the production of the tardigrade intrinsically disordered protein (TDP) "CAHS 94205". This is one of the several heat soluble proteins found in the tardigrades capable of protecting other fragile biological parts upon desiccation by generating a crystal matrix. Therefore, this tardigrade protein can be combined with other perishable biological material (e.g. proteins) to increase its stability after dehydration or even enable the possibility of stabilizing material with just a previous drying step. The sequence encoding for the gene is preceded by a T7 promoter BBa_I712074 and an RBS BBa_K2306014, which allows inducible overexpression of the protein with IPTG; and followed by a double terminator consisting of a rrnB T1 terminator and T7Te terminator BBa_B0015. This was the part employed in TU Delft's project for the expression and production of CAHS 94205. More information about our project can be found on our [http://2017.igem.org/Team:TUDelft/Results#TDP results page].
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 549
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 268
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
A T7 promoter and a strong RBS were chosen to the end of maximizing the production of the protein and therefore enable the subsequent characterization experiments
Source
Registry of Standard Biological Parts, Boothby et al., 2017, Molecular Cell 65, 975–984 March 16, 2017; 2017 Elsevier Inc.
References
Boothby et al., 2017