Difference between revisions of "Part:BBa K260017:Design"

 
 
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__NOTOC__
 
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<partinfo>BBa_K260017 short</partinfo>
 
<partinfo>BBa_K260017 short</partinfo>
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===Design Notes===
 
===Design Notes===
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A strong promoter was necessary and we use BBa_J23100. An appropriate reading frame for the F3 sites had to be found to prevent premature Stop-codons and hydrophobic amino acid residues. Further, the genes encoding ZeoR and mRFP1 were codon optimised for expression in both E. coli and H. sapiens. For potential use in H. sapiens, the CDS were also made free of polyA sites, exon junctions and CpG dinucleotides where possible. The leading alanine of ZeoR was omitted to have exactly 500 bp between both F3 sites.
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A strong promoter was necessary and we use [[Part:BBa_J23100|BBa_J23100]]. An appropriate reading frame for the F3 sites had to be found to prevent premature Stop-codons and hydrophobic amino acid residues. Further, the genes encoding ZeoR and mRFP1 were codon optimised for expression in both E. coli and H. sapiens. For potential use in H. sapiens, the CDS were also made free of polyA sites, exon junctions and CpG dinucleotides where possible. The leading alanine of ZeoR was omitted to have exactly 500 bp between both F3 sites.
  
  
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This part was synthesised by "Mr Gene" (Geneart) and is available in the following plasmid backbones:
 
This part was synthesised by "Mr Gene" (Geneart) and is available in the following plasmid backbones:
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pMA-RQ-@TetFlp (BBa_K260009)
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pTetFlp (BBa_K260002)
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[[Part:BBa_K260009|BBa_K260009]]: pMA-RQ-@TetFlp
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pRhaFlp (BBa_K260003)
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 +
[[Part:BBa_K260002|BBa_K260002]]: pTetFlp
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[[Part:BBa_K260003|BBa_K260003]]: pRhaFlp
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 +
 
  
 
===References===
 
===References===

Latest revision as of 08:50, 18 October 2009

P_F3_ZeoR_F3_RFP. [FLP]->RFP


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 420


Design Notes

A strong promoter was necessary and we use BBa_J23100. An appropriate reading frame for the F3 sites had to be found to prevent premature Stop-codons and hydrophobic amino acid residues. Further, the genes encoding ZeoR and mRFP1 were codon optimised for expression in both E. coli and H. sapiens. For potential use in H. sapiens, the CDS were also made free of polyA sites, exon junctions and CpG dinucleotides where possible. The leading alanine of ZeoR was omitted to have exactly 500 bp between both F3 sites.


Source

This part was synthesised by "Mr Gene" (Geneart) and is available in the following plasmid backbones:

BBa_K260009: pMA-RQ-@TetFlp

BBa_K260002: pTetFlp

BBa_K260003: pRhaFlp


References