Difference between revisions of "Part:BBa K802003"
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===Sequencing of the modified regions ===
 
===Sequencing of the modified regions ===
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===Usage and Biology===
 
  
  
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===Usage and Biology===
  
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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. 
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===Functional Parameters===
 
===Functional Parameters===
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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 ????
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Revision as of 22:45, 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:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal prefix found in sequence at 6506
    Illegal suffix found in sequence at 1
  • 12
    INCOMPATIBLE 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
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 6506
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found in sequence at 6506
    Illegal suffix found in sequence at 2
  • 25
    INCOMPATIBLE 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
  • 1000
    INCOMPATIBLE 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.

Antibiotic resistance

Sequencing of the modified regions

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 ????