Plasmid
Part:BBa_K4404010:Design
Designed by: Julia Fricke Group: iGEM22_Goettingen (2022-10-09)
acsB2 from C. ljungdahlii under the control of a PbgaL promoter from C. perfringens
Assembly Compatibility:
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 2623
Illegal EcoRI site found at 6098
Illegal XbaI site found at 6125
Illegal SpeI site found at 1777
Illegal PstI site found at 542
Illegal PstI site found at 5988
Illegal PstI site found at 6164 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 2623
Illegal EcoRI site found at 6098
Illegal NheI site found at 2230
Illegal NheI site found at 6343
Illegal SpeI site found at 1777
Illegal PstI site found at 542
Illegal PstI site found at 5988
Illegal PstI site found at 6164
Illegal NotI site found at 6067 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 2623
Illegal EcoRI site found at 6098
Illegal BglII site found at 6152
Illegal BamHI site found at 2214
Illegal BamHI site found at 6119
Illegal XhoI site found at 97
Illegal XhoI site found at 6156 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 2623
Illegal EcoRI site found at 6098
Illegal XbaI site found at 6125
Illegal SpeI site found at 1777
Illegal PstI site found at 542
Illegal PstI site found at 5988
Illegal PstI site found at 6164 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 2623
Illegal EcoRI site found at 6098
Illegal XbaI site found at 6125
Illegal SpeI site found at 1777
Illegal PstI site found at 542
Illegal PstI site found at 5988
Illegal PstI site found at 6164
Illegal NgoMIV site found at 3537
Illegal NgoMIV site found at 5444
Illegal NgoMIV site found at 5454
Illegal NgoMIV site found at 5580 - 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 5196
Illegal SapI.rc site found at 5551
Illegal SapI.rc site found at 5922
Design Notes
It is important to produce inserts with specific restriction enzymes in order to secure a robust implementation into a vector system.
Source
The constitutive promoter originated from C. perfringens, while acsB2 was amplified from genomic DNA of Clostridium ljungdahlii with specific primers by PCR. The pMTL83151 based on the ClosTron plasmid system and was modified by Prof. Peter Dürre (department of microbiology and biotechnology).