Difference between revisions of "Part:BBa K1898250:Design"
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Our assembly method BioBrick RFC[10] shifted the open reading frame and created a stop codon in between GSR and 10x histidine tag. This is showed in this part's main page under sequencing. | Our assembly method BioBrick RFC[10] shifted the open reading frame and created a stop codon in between GSR and 10x histidine tag. This is showed in this part's main page under sequencing. | ||
The sequence was sent to mutagenesis to add two base pairs in between the two genes. | The sequence was sent to mutagenesis to add two base pairs in between the two genes. | ||
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===Source=== | ===Source=== | ||
Revision as of 16:09, 18 October 2016
Strong promoter + Strong RBS + GSR + 10x Histidine tag + Double terminator
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 804
Illegal BamHI site found at 1488 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 89
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI site found at 1199
Illegal SapI.rc site found at 1607
Design Notes
We did five silent point mutations to the GSR sequence to remove the two Pst1 and three EcoR1 internal cutting sites. The sequence was sent out for mutagenesis.
Primers were designed to remove the stop codon from GSR and to move the cDNA into iGEM Biobrick.
Our assembly method BioBrick RFC[10] shifted the open reading frame and created a stop codon in between GSR and 10x histidine tag. This is showed in this part's main page under sequencing. The sequence was sent to mutagenesis to add two base pairs in between the two genes.
Source
the cDNA of GSR was ordered from OriGene. Mutagensis were performed by MissionBiotech. Primers were synthesized by Tri-I Biotech.