Difference between revisions of "Part:BBa K802003"
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| − | The shuttle vector is Ampicillin resistant in E. coli and Erythromycin resistant in B. subtilis. It is a low plasmid copy number in B. subtilis and a high copy number in E.coli. | + | The shuttle vector of 6,5kb is Ampicillin resistant in E. coli and Erythromycin resistant in B. subtilis. It is a low plasmid copy number in B. subtilis and a high copy number in E.coli. It has a polylinker containing all 4 of the iGEM sites. |
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Revision as of 15:59, 24 September 2012
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 low plasmid copy number in B. subtilis and a high copy number in E.coli. It has a polylinker containing all 4 of the iGEM sites.
Sequence and Features
Assembly Compatibility:
- 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 3,7XE06 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 81 cells/µg. The transformed cells were selected on LB media supplemented with Erythromycin at a concentration of 15µg/mL.