Primer
HE RBS

Part:BBa_K1539021:Design

Designed by: Coleen Tran   Group: iGEM14_GeorgiaTech   (2014-10-10)


HE RBS Primer


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal prefix found in sequence at 3
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 3
    Illegal NotI site found at 9
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 3
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found in sequence at 3
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal prefix found in sequence at 3
    Illegal XbaI site found at 18
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Needed to design the primer so that the annealing temperature for PCR was 60 degrees Celsius, which is the same annealing temperature as the VR primer.

The primer designed to maintain the EcoRI and XbaI sites but still insert an RBS.

The RBS was chosen from the Salis Lab RBS Library with a theoretical translation initiation rate of 0.914 compared to the Shine-Delgarno sequence.

Source

Primer ordered from IDT

References