Part:BBa_K424017:Design
Test plataform for rhamnosyltransferase BioBrick (Rh1AB_BB) expression in E. coli
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 220
Illegal BamHI site found at 780
Illegal XhoI site found at 956
Illegal XhoI site found at 2242 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 1135
Illegal NgoMIV site found at 1856
Illegal NgoMIV site found at 1969 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 445
Illegal BsaI site found at 1485
Illegal BsaI.rc site found at 629
Illegal BsaI.rc site found at 3135
Design Notes
We were able to ligate successfully the promotor with the RBS into a plasmid backbone and the reporter with the terminator; and we get the rhamnosiltransferase 1 gene enzyme mutated to be compatible with the Assembly Standard 10. We need to ligate all the parts to test our plataform for rhamnosyltransferase expression.
Source
The promoter, RBS, GFP reporter and the terminator were given to us as a genetic tool kit by iGEM. The Standard Rh1AB part was designed by Team Panama 2010. The rhamnosyltransferase 1 complex (Rh1AB) was isolated from a Pseudomonas aeruginosa and mutated by a mutagenesis protocol from Stratagene to be compatible with the Assembly Standard Protocol 10.
References
Assembly Standard Protocol 10, www.parts.igem.org
QuikChange Lightning Multi Site-Directed Mutagenesis Kit Instruction manual, catalog #210515 from Stratagene.