Part:BBa_K802004
Shuttle vector for E. coli and B. subtilis
The shuttle vector of 6,5kb is Ampicillin resistant in E. coli and Erythromycin resistant in B. subtilis. It is a high plasmid copy number in both B. subtilis and E.coli. It has a polylinker containing all 4 of the iGEM sites.
Usage and Biology
It can be used to transform E. coli and Bacillus strains. However, it is highly recommended to amplify the plasmid before transforming a Bacillus strain because the transformation yield is low compared to E. coli.
Functional Parameters
The tests showed that in the NM522 E. coli strain there is bacterial growth in LB media having an Ampicillin concetration up to 1,2 mg/mL. Concerning the 168 B. subtilis strain transformed with this plasmid the maximal concentration of Erythromycin at wich there is bacterial growth is ????
- 10INCOMPATIBLE WITH RFC[10]Illegal prefix found in sequence at 6506
Illegal suffix found in sequence at 1 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 6506
Illegal NheI site found at 3377
Illegal SpeI site found at 2
Illegal PstI site found at 16
Illegal NotI site found at 9
Illegal NotI site found at 6512 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 6506
- 23INCOMPATIBLE WITH RFC[23]Illegal prefix found in sequence at 6506
Illegal suffix found in sequence at 2 - 25INCOMPATIBLE WITH RFC[25]Illegal prefix found in sequence at 6506
Illegal XbaI site found at 6521
Illegal SpeI site found at 2
Illegal PstI site found at 16 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 3203
Illegal BsaI.rc site found at 5190
Characterization
Transformation yield
In the NM522 E.coli strain the average yield was 3,3XE06 cells/µg.The transformed cells were selected on LB media supplemented with Ampicillin at a concentration of 100µg/mL.
In the BS168 B.subtilis strain the maximum yield was 70 cells/µg. The transformed cells were selected on LB media supplemented with Erythromycin at a concentration of 15µg/mL.
Antibiotic resistance
Sequencing of the modified regions
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