Difference between revisions of "Part:BBa K1582021:Design"
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===Design Notes=== | ===Design Notes=== | ||
| − | + | '''3.1 Primer Design'''<br> | |
| − | + | To make FsC and sJanus-m into a fusion protein, we use overlap PCR and use 30bp as the overlapping sequence. We choose XbaI and PstI as restriction sites and add protective bases before two sites. Primers designed are as follows:<br> | |
| − | + | F1: 5’- GAATTCGCGGCCGCTTCTAGATGCTGCCTACTTCTAACCCT-3’<br> | |
| + | R1: 5’- AGAACCTCCACCGCCAGAACCTCCTCCACCAGCAGAACCACGGAC-3’<br> | ||
| + | F2: 5’- GGTGGAGGAGGTTCTGGCGGTGGAGGTTCTAGCAACGGCAACGGC-3‘<br> | ||
| + | R2: 5’-CCCTTTTTTGCCGGACTGCAGCGGCCGCTACTAGTATCAAGCACCGACGGC-3’<br> | ||
| + | '''3.2 Linker Design'''<br> | ||
| + | GGGGSGGGGS is a mature linker and has been used in many researches. So we choose it and it is codon optimized into ACCTACTCTGACAACCGTTACGGTGGTACCTGCGACCCGGACGGTTGCGACTGGAACCCGTACCGTCTGGGTAAC.<br> | ||
===Source=== | ===Source=== | ||
Latest revision as of 03:03, 25 September 2015
FsC+sJanus-m Fusion Protein
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NotI site found at 379
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 525
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 907
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 301
Illegal SapI site found at 916
Design Notes
3.1 Primer Design
To make FsC and sJanus-m into a fusion protein, we use overlap PCR and use 30bp as the overlapping sequence. We choose XbaI and PstI as restriction sites and add protective bases before two sites. Primers designed are as follows:
F1: 5’- GAATTCGCGGCCGCTTCTAGATGCTGCCTACTTCTAACCCT-3’
R1: 5’- AGAACCTCCACCGCCAGAACCTCCTCCACCAGCAGAACCACGGAC-3’
F2: 5’- GGTGGAGGAGGTTCTGGCGGTGGAGGTTCTAGCAACGGCAACGGC-3‘
R2: 5’-CCCTTTTTTGCCGGACTGCAGCGGCCGCTACTAGTATCAAGCACCGACGGC-3’
3.2 Linker Design
GGGGSGGGGS is a mature linker and has been used in many researches. So we choose it and it is codon optimized into ACCTACTCTGACAACCGTTACGGTGGTACCTGCGACCCGGACGGTTGCGACTGGAACCCGTACCGTCTGGGTAAC.