Part:BBa_K1033259:Design
Toxin-antitoxin system from lactobacillus plantarum
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 71
Illegal SpeI site found at 447 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 71
Illegal SpeI site found at 447 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 71
- 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 71
Illegal SpeI site found at 447 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 71
Illegal SpeI site found at 447 - 1000COMPATIBLE WITH RFC[1000]
Design Notes
The system has been amplified without RBS and its natural promotor. Since that promotor has not been characterized as far as we know, several promoters will probably have to be tried to find one that provides stability without adding to much cost of expressing the toxin-antitoxin. There is also a version available with the natural promoter and RBS. The part still contains two illegal restriction sites for EcorI and SpeI. However since it doesn't matter where in your construct you put the system it can still be assembled with XbaI and PstI.
Source
The Lactobacillus plantarum NC7 plasmid p256.
References
Plasmid p256 from Lactobacillus plantarum represents a new type of replicon in lactic acid bacteria, and contains a toxin–antitoxin-like plasmid maintenance system, Microbiology, Elisabeth Sorvig et al. September 30 2004