Difference between revisions of "Part:BBa K2938014: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)
  
 
===Source===
 
===Source===

Latest revision as of 20:30, 9 October 2019


Cry4Ba (HA tag) + deGFP Device


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 1859
    Illegal XbaI site found at 3597
    Illegal SpeI site found at 2548
    Illegal PstI site found at 363
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 56
    Illegal NheI site found at 105
    Illegal NheI site found at 247
    Illegal SpeI site found at 2548
    Illegal PstI site found at 363
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 4330
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 1859
    Illegal XbaI site found at 3597
    Illegal SpeI site found at 2548
    Illegal PstI site found at 363
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 1859
    Illegal XbaI site found at 3597
    Illegal SpeI site found at 2548
    Illegal PstI site found at 363
    Illegal NgoMIV site found at 2671
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 653


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