Composite
Part:BBa_K2924052:Design
Designed by: Melanie Sbielut Group: iGEM19_Duesseldorf (2019-10-16)
Hpall + SPNprE + alpha s2 + fd terminator
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 1124
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 554
Design Notes
Expression system for the expression and secretion of the protein in B. subtilis
Source
PBSMul1-SPNprE plasmid was kindly provided by Dr. Andreas Knapp from the research center Jülich. The DNA sequence was acquired by reverse-translating the protein sequence of a-s1-casein. a-s1-casein was commercial synthesized by GenScript.
References
Brockmeier, U., et al. (2006). "Systematic screening of all signal peptides from Bacillus subtilis: a powerful strategy in optimizing heterologous protein secretion in Gram-positive bacteria." J Mol Biol 362(3): 393-402.
Knapp, A., et al. (2017). "Activity-independent screening of secreted proteins using split GFP." J Biotechnol 258: 110-116.