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Part:BBa_K801999:Design

Designed by: Volker Morath   Group: iGEM12_TU_Munich   (2012-08-19)
Revision as of 10:13, 13 September 2012 by VolkerMorath (Talk | contribs) (→‎References)

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:



TUM12-Test page1.png


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

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:

  • Genesequence derived from Aequorea victoria
  • Codonoptimized 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 Chromophor)]

Seqeuence references:

  • [http://www.ncbi.nlm.nih.gov/nuccore/X83959.1 GeneBank: 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]