Coding

Part:BBa_K1539002:Design

Designed by: Coleen Tran   Group: iGEM14_GeorgiaTech   (2014-10-09)
Revision as of 20:58, 1 November 2014 by Stassoulas3 (Talk | contribs) (→‎Design Notes)


LPromoter->LRBS->mCherry


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

mCherry from BBa_J0650 was amplified with BBa_K1539034. This product were Dpn1 digested and then PCR cleaned up. The product was then PCR re-amplified with BBa_K1539089, digested with XbaI and PstI and ligated into a similarly digested pSB1c3 backbone. Amplification of each step was confirmed by gel electrophresis producing bands approximately 900-950 bp (~700 bp for mCherry, 30 bp for RBS, 60 bp for promoter, and ~150 in extra sequence added by codons beyond the suffix to the reverse sequencing primer).

Source

mCherry from BBa_J06504 RBS and promoter from created primers

References