Difference between revisions of "Part:BBa K801999:Design"
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<br>'''Keywords:'''
 
<br>'''Keywords:'''
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<!--These keywords are necessary to find your part using a fulltext sarch.-->
 
<!--keyword_1, keyword_2, keyword_3, keyword_4, keyword_5-->
 
<!--keyword_1, keyword_2, keyword_3, keyword_4, keyword_5-->
  
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<!--*Part was truncated upstream/downstream compared to template in order to ???-->
 
<!--*Part was truncated upstream/downstream compared to template in order to ???-->
 
<!--*The correctness of the part was checked by sequencing./The correctness of the part was checked by test digestion using ?enzyme1? and ?enzyme2?/The correctness of the part was not checked.-->
 
<!--*The correctness of the part was checked by sequencing./The correctness of the part was checked by test digestion using ?enzyme1? and ?enzyme2?/The correctness of the part was not checked.-->
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'''Backbone:'''<br>
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<!--*Backbone name: ?backbone_name?-->
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<!--*This particular backbone was chosen because ???-->
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<!--*Resistance: ???-->
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<!--*Copynumber: low/medium/high-->
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'''Chassis compability:'''<br>
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<!--*Part was designed for ?chassi?-->
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<!--*Part should also work in ?other_chassi?.-->
  
 
'''Protein coding:'''<br>
 
'''Protein coding:'''<br>
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<!--*The BioBrick does not encode a protein" or ?Name_of_gene_product? [Nucleotide 1 to ???]-->
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<!--*Protein: ?Name_of_gene_product? [Nucleotide 1 to ???]-->
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<!--*The protein has the amino acid replacements ???99??? to ???.-->
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<!--*The protein has the amino acid replacements ???99??? to ???99???.-->
 
<!--*The protein encoded is posttranslationally modified by ???.-->
 
<!--*The protein encoded is posttranslationally modified by ???.-->
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<!--*Tag: n-terminally fused/c-terminally fused His5/His6/Strep/Flag/???-->
  
 
'''Enzymatic activity:'''
 
'''Enzymatic activity:'''
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'''Cytotoxicity:'''
 
'''Cytotoxicity:'''
 
<!--none/not known/cytotoxic for ''organism name''-->
 
<!--none/not known/cytotoxic for ''organism name''-->
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'''Intellectual Property information:'''
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<!--The part contains seqeunces that are protected by the patent ???-->
  
 
===Source===
 
===Source===
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'''Source:'''<br>
 
'''Source:'''<br>
 
<!--*Amplified from commercial system: plasmid name, system name, company name<br>-->
 
<!--*Amplified from commercial system: plasmid name, system name, company name<br>-->
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<!--*Genesequence derived from ''?organism_name?''-->
 
<!--*Genesequence derived from ''?organism_name?''-->
 
<!--*Codonoptimized for ''?organism_name?''-->
 
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===References===
 
===References===

Revision as of 15:59, 16 September 2012

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:

Backbone:

Chassis compability:

Protein coding:

Enzymatic activity:

Cytotoxicity:

Intellectual Property information:

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 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]