Difference between revisions of "Part:BBa K2938018:Design"

 
Line 7: Line 7:
  
 
===Design Notes===
 
===Design Notes===
−
Hadar writes
+
You can cut out any subunit using an upstream RBS and downstream of the Subunit restrictions sites.
 +
 
 +
The order of the subunits downstream of the promoter was determined by the importance of the subunits to the overall toxicity of the plasmid. Due to the usage of one promoter, the amount of mRNA of the last subunits that will form will decrease.
 +
 
 +
The last gene in the polycistronic plasmid is deGFP without a tag.
 +
 
 +
The presence of the GFP signal proves the expression of the previous subunits on the plasmid and it allows the detection of the transgenic bacteria easily (in mosquitoes)
  
  

Revision as of 20:30, 9 October 2019


Polycistronic BTI Toxin


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1846
    Illegal EcoRI site found at 2142
    Illegal EcoRI site found at 6316
    Illegal EcoRI site found at 6627
    Illegal XbaI site found at 3820
    Illegal XbaI site found at 5510
    Illegal SpeI site found at 958
    Illegal SpeI site found at 4509
    Illegal PstI site found at 2324
    Illegal PstI site found at 5976
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1846
    Illegal EcoRI site found at 2142
    Illegal EcoRI site found at 6316
    Illegal EcoRI site found at 6627
    Illegal NheI site found at 56
    Illegal NheI site found at 105
    Illegal NheI site found at 2093
    Illegal NheI site found at 2208
    Illegal SpeI site found at 958
    Illegal SpeI site found at 4509
    Illegal PstI site found at 2324
    Illegal PstI site found at 5976
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1846
    Illegal EcoRI site found at 2142
    Illegal EcoRI site found at 6316
    Illegal EcoRI site found at 6627
    Illegal BglII site found at 6274
    Illegal BamHI site found at 5653
    Illegal XhoI site found at 7655
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1846
    Illegal EcoRI site found at 2142
    Illegal EcoRI site found at 6316
    Illegal EcoRI site found at 6627
    Illegal XbaI site found at 3820
    Illegal XbaI site found at 5510
    Illegal SpeI site found at 958
    Illegal SpeI site found at 4509
    Illegal PstI site found at 2324
    Illegal PstI site found at 5976
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1846
    Illegal EcoRI site found at 2142
    Illegal EcoRI site found at 6316
    Illegal EcoRI site found at 6627
    Illegal XbaI site found at 3820
    Illegal XbaI site found at 5510
    Illegal SpeI site found at 958
    Illegal SpeI site found at 4509
    Illegal PstI site found at 2324
    Illegal PstI site found at 5976
    Illegal NgoMIV site found at 4632
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 2614


Design Notes

You can cut out any subunit using an upstream RBS and downstream of the Subunit restrictions sites.

The order of the subunits downstream of the promoter was determined by the importance of the subunits to the overall toxicity of the plasmid. Due to the usage of one promoter, the amount of mRNA of the last subunits that will form will decrease.

The last gene in the polycistronic plasmid is deGFP without a tag.

The presence of the GFP signal proves the expression of the previous subunits on the plasmid and it allows the detection of the transgenic bacteria easily (in mosquitoes)


Source

Gibson Assembly

References