Coding

Part:BBa_K1484103:Design

Designed by: Guy Lewy   Group: iGEM14_Cambridge-JIC   (2014-10-08)

Benzalacetone Synthase, compatible with MoClo


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 1080
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 384
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1
    Illegal BsaI.rc site found at 1169


Design Notes

Benzalacetone Synthase has been designed to be compatible with GoldenGate standards. This allows the part to be used in MoClo (ModularCloning) or GB2 (goldenbraid2) assembly method. These cloning techniques are based on based on Type IIS restriction enzymes that enable parallel assembly of multiple parts in a one-pot, one-step reaction and are becoming widely adopted across the field of Synthetic Biology.

To be in the GoldenGate format, a level 0 CDS part must: start with -the BsaI_F recognition site 

  GGTCTC

-one non-specific nucleotide 

  A

-4bp fusion sequence (for CDS this is AATG) 

  AATG

and finish after the cds with -4bp fusion sequence (for CDS this is GCTT) 

  GCTT

-one non-specific nucleotide 

  T

-the BsaI_R recognition site reversed 

  GAGACC


The BAS GoldenGate biobrick reads: Prefix_GGTCTC A AATG ___BAS(without atg)___GCTT T GAGACC_Suffix

Source

Cloned from the genome of Rheum palmatum (Chinese rhubarb) by the lab of Ikuro Abe at the Graduate School of Pharmaceutiacal Sciences, Tokyo

References