Difference between revisions of "Part:BBa K260016:Design"
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__NOTOC__ | __NOTOC__ | ||
<partinfo>BBa_K260016 short</partinfo> | <partinfo>BBa_K260016 short</partinfo> | ||
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===Source=== | ===Source=== | ||
| − | This part was synthesised by "Mr Gene" (Geneart) and is available in the original pMA-BsdR-@CC2FosMB plasmid backbone ([[Part:BBa_K260010|]]), but also pCC2FosMB [[Part:BBa_K260001|]], which is already half-way towards constructing the full P_FRT_dhfr_TT_FRT_GFP reporter. | + | This part was synthesised by "Mr Gene" (Geneart) and is available in the original pMA-BsdR-@CC2FosMB plasmid backbone ([[Part:BBa_K260010|BBa_K260010]]), but also pCC2FosMB [[Part:BBa_K260001|BBa_K260001]], which is already half-way towards constructing the full P_FRT_dhfr_TT_FRT_GFP reporter. |
===References=== | ===References=== | ||
Revision as of 15:56, 18 October 2009
TT_FRT_GFP. (double terminator, FRT site, optimised GFP)
Assembly Compatibility:
- 10INCOMPATIBLE WITH RFC[10]Illegal XbaI site found at 144
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal XbaI site found at 144
- 25INCOMPATIBLE WITH RFC[25]Illegal XbaI site found at 144
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 809
Design Notes
A strong promoter was necessary and we use BBa_J23100. An appropriate reading frame for the FRT site had to be found to prevent premature Stop-codons and hydrophobic amino acid residues. Further, the gene encoding GFP (mut3b) was 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.
Source
This part was synthesised by "Mr Gene" (Geneart) and is available in the original pMA-BsdR-@CC2FosMB plasmid backbone (BBa_K260010), but also pCC2FosMB BBa_K260001, which is already half-way towards constructing the full P_FRT_dhfr_TT_FRT_GFP reporter.