Plasmid

Part:BBa_K1391013:Design

Designed by: Shinjini Saha   Group: iGEM14_MIT   (2014-10-17)


pENTR_PirB_TCS_Gal4VP16


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 244
    Illegal EcoRI site found at 1735
    Illegal PstI site found at 1324
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 244
    Illegal EcoRI site found at 1735
    Illegal PstI site found at 1324
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 244
    Illegal EcoRI site found at 1735
    Illegal BamHI site found at 53
    Illegal XhoI site found at 3005
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 244
    Illegal EcoRI site found at 1735
    Illegal PstI site found at 1324
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 244
    Illegal EcoRI site found at 1735
    Illegal PstI site found at 1324
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 2924
    Illegal SapI.rc site found at 2062


Design Notes

This part was created using scarless golden gate assembly. This basic part is flanked by L1 and L2 sites and can be easily cloned into an entry vector using an LR reaction. A promoter can be easily inserted in front of this part in a one pot LR reaction with a promoter flanked by L4 and R1 sites cloned into a backbone that has a negative selection marker between R4 and R2 sites. This part adheres to RFC 65 for recombination based cloning of mammalian parts.

Source

Composite: Mouse, tobacco etch virus

References