Difference between revisions of "Part:BBa K627007:Design"
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===Source=== | ===Source=== | ||
| − | + | The gene-III-protein is a coat protein from the filamentous bacteriophage M13. | |
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===References=== | ===References=== | ||
Revision as of 19:35, 21 September 2011
Fusion of c-myc-tag and gene III
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
Gene III was amplified from the vector pak100 bla KDIR using the following primer
Forward: TAAGCTTCTAGATGGCCGGCGAGCAGAAGCTGATCTCTGAGGAAGACCTGGGTGGTGGCTCTGGTTCC
Reverse: TGCTTAGACGTCCTGCAGCGGCCGCTACTAGTATTAACCGGTAGACTCCTTATTACGCAGTA
Because c-myc-gene III is a fusion part (RFC25), no start codon is needed. Before the c-myc sequence a NgoMIV restriction site is located which has compatible overhangs to AgeI restriction sites. The ligation of AgeI and NgoMIV overhangs will result in a scar coding for threonine and glycine. So genes of interest containing AgeI restriction sites can be easily fused to gene III without frame shift. The gene III sequence contains no stop codon because there is one located between AgeI and SpeI recognition site directly merged behind gene III sequence.
Source
The gene-III-protein is a coat protein from the filamentous bacteriophage M13.