Composite

Part:BBa_K1582021:Design

Designed by: LIU JIASHU   Group: iGEM15_Tianjin   (2015-08-07)
Revision as of 03:03, 25 September 2015 by Sherry222 (Talk | contribs) (→‎Design Notes)

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FsC+sJanus-m Fusion Protein


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NotI site found at 379
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 525
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 907
  • 1000
    INCOMPATIBLE 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.

Source

References