Coding

Part:BBa_K2524002:Design

Designed by: Holly Bowman   Group: iGEM17_Georgia_State   (2017-09-29)


Limulus Clotting Factor C-hCG fusion protein


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The basic mechanism for overlap begins with a PCR to generate two fragments (Block 1 and Block 2) that have ends modified by mispriming so that they have homologous regions. Through mixture, denaturation, and annealing of Block 1 5'-end of the top strand will anneal onto the 3'-end of the bottom strand of Block 2. Extension of the overlap product is used to form the recombinant product (the overlapping ends of the fragments are created by the forward primers and reverse complement primers to match the individual fragments). The method can be repeated to add together more than two DNA fragments (Block 3 and Block 4 + hCG-beta subunit). Together, all 5 g-blocks and a total of 10 primers would be joined via overlap extension PCR. If this joining of LFC blocks 1-4 and hCG-beta subunit block 5 is successful, the final product of Limulus Clotting Factor C + hCG-beta subunit sequence would be used as our complete detection system.

Source

http://www.ebi.ac.uk/ena/data/view/BAA14316


References