Part:BBa_K1152005:Design
Expression cassette for NRPS synthesizing a Phe-Orn-Leu-Tripeptide
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 6617
Illegal BsaI.rc site found at 2745
Illegal BsaI.rc site found at 5490
Illegal SapI.rc site found at 7734
Illegal SapI.rc site found at 7971
Design Notes
BBa_K1152004 was assembled via Gibson Cloning out of three fragments. The modules that were required for the NRPS were amplified from B. parabrevis. There were no illegal restiction sites, neither in TycAdCom, nor in TycC5 and TycC6. As TycA does not need protein-protein interaction, but is covalently bound to the subsequent module, the Com-doamin was not included in the costruct.
Source
Coding regions were amplified from genome of B. parabrevis that was sent to us by the lab of Prof. Marahiel from Marburg.
References
1. Marahiel MA, Stachelhaus T, Mootz HD (1997) Modular Peptide Synthetases Involved in Nonribosomal Peptide Synthesis. Chem Rev 97:2651–2674.
2. Mootz HD, Schwarzer D, Marahiel MA (2000) Construction of hybrid peptide synthetases by module and domain fusions. Proc Natl Acad Sci USA 97: 5848–5853.
3. Mootz HD, Marahiel MA (1997) The tyrocidine biosynthesis operon of Bacillus brevis: complete nucleotide sequence and biochemical characterization of functional internal adenylation domains. JBacteriol 179: 6843–6850.
4. Finking R, Marahiel MA (2004) Biosynthesis of nonribosomal peptides. Annu Rev Microbiol 58: 453–488.