Difference between revisions of "Part:BBa J100069"

Latest revision as of 18:11, 2 July 2012

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

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 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 ####), but the primers were were made to use with un-optimized superfolder GFP ([https://parts.igem.org/Part:BBa_I746916 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).
+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 ([https://parts.igem.org/Part:BBa_J100070 Part: Bba_J100070]), but the primers were were made to use with un-optimized superfolder GFP ([https://parts.igem.org/Part:BBa_I746916 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).
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