Device

Part:BBa_K2938016:Design

Designed by: Assaf Vital   Group: iGEM19_BGU_Israel   (2019-10-09)


Cry11Aa (Strep tag) + P20 (His tag) + deGFP Device


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1870
    Illegal EcoRI site found at 2426
    Illegal XbaI site found at 2116
    Illegal XbaI site found at 2761
    Illegal SpeI site found at 982
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1870
    Illegal EcoRI site found at 2426
    Illegal NheI site found at 56
    Illegal NheI site found at 105
    Illegal SpeI site found at 982
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1870
    Illegal EcoRI site found at 2426
    Illegal XhoI site found at 3494
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1870
    Illegal EcoRI site found at 2426
    Illegal XbaI site found at 2116
    Illegal XbaI site found at 2761
    Illegal SpeI site found at 982
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1870
    Illegal EcoRI site found at 2426
    Illegal XbaI site found at 2116
    Illegal XbaI site found at 2761
    Illegal SpeI site found at 982
  • 1000
    COMPATIBLE WITH RFC[1000]


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