Part:BBa_J100069

Superfolder GFP

This part is the coding sequence for superfolder GFP with a BsaI restriction site at the beginning for use with the Golden Gate Assembly method. The BsaI site cuts forward cleaving the sequence right before the start codon. The sticky word made is ATGC (the start codon and the first base pair of the sequence).

The superfolder GFP coding sequence has been mostly codon optimized for E. coli. We made this part using PCR. The template DNA was a fully optimized superfolder GFP coding sequence (Part: Bba_J100070), but the primers were were made to use with un-optimized superfolder GFP (Part:Bba_I746916). For this reason the 4th, 6th and 8th codons (coding for Glycine, Glutamic acid, and Phenylalanine respectively) as well as the first stop codon are not optimized for E. coli. There was also one A to G point mutation in the 34th codon (GAA to GAG), but it does not change the amino acid sequence (the 34th amino acid is still Glutamic acid).

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal prefix found in sequence at 1
  Illegal suffix found in sequence at 750
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 1
  Illegal SpeI site found at 751
  Illegal PstI site found at 765
  Illegal NotI site found at 7
  Illegal NotI site found at 758
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 1
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal prefix found in sequence at 1
  Illegal suffix found in sequence at 751
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal prefix found in sequence at 1
  Illegal XbaI site found at 16
  Illegal SpeI site found at 751
  Illegal PstI site found at 765
  Illegal AgeI site found at 54
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 23
  Illegal SapI.rc site found at 42