Part:BBa_K801999:Design

Test page for standardized BioBrick part descriptions

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]

Keywords:

Abbreviations:

Design Notes

Other versions of this BioBrick:

Cloning details:

Protein coding:

Enzymatic activity:

Cytotoxicity:

Source

Source:

Organism:

References

Literature references:

Sequence references:

Structure reference:

Test page for standardized BioBrick part descriptions

Keywords: fluorescent, reporter, chromophore, luminescence, bioluminescence, photoprotein

Used abbreviations:

• GFP = Green Fluorescent Protein

Design Notes

Other versions of this BioBrick:

• The BioBrick BBa_I757008 encodes a yellow fluorescent protein (mVenus) derived from GFP
• The BioBrick BBa_I757008 encodes GFP in RFC25 for protein fusions

Cloning details:

• Part was designed in RFC10
• Mutation G381A to delete XbaI restriction site
• The correctness of the part was checked by sequencing

Protein coding:

• Green Fluorescent Protein [Nucleotide 1 to 714]
• The protein has the amino acid replacements Ser65Thr in order to increase fluorescence, photostability and to shift the major excitation peak to 488 nm. (see Heim et al., 1995)
• The protein encoded is posttranslationally modified by a cross-link between Ser65 and Gly67 to form the chromophore.

Enzymatic activity: none

Cytotoxicity: none

Source

Source:

• Amplified from plasmid: pSB1C3-GFP-generator, provided by Osamu Shimomura, Boston University School of Medicine, USA

Forward Primer:
5'- ATGATGATGATG - 3'
Reverse Primer:
5'- ATGATGATGATG - 3'

Organism:

• Sequence derived from Aequorea victoria
• Codon optimized for Escherichia coli

References

Literature references:

• [http://www.ncbi.nlm.nih.gov/pubmed/20010584 Pubmed: Prasher, 1995: Using GFP to see the light.(Review)]
• [http://www.ncbi.nlm.nih.gov/pubmed/7854443 Pubmed: Heim, 1995: Improved green fluorescence.(Reference for Chromophore)]

Sequence references:

• [http://www.ncbi.nlm.nih.gov/nuccore/X83959.1 GenBank: A.victoria mRNA for green fluorescent protein (ID:gfp1)]
• [http://www.ebi.ac.uk/interpro/IEntry?ac=IPR011584 Interpro: Green fluorescent protein-related ]
• [http://www.uniprot.org/uniprot/P42212 Uniprot: Green fluorescent protein]
• [http://pfam.sanger.ac.uk/family/PF01353 Pfam: Green fluorescent protein]

Structure reference:

• [http://www.rcsb.org/pdb/explore/explore.do?structureId=1EMA PDB: Green Fluorescent Protein from Aequorea victoria]