Coding
linker

Part:BBa_K365005:Design

Designed by: STIEFEL Fabian & KALCHSCHMIDT Jens Sebastian   Group: iGEM10_ESBS-Strasbourg   (2010-09-14)

20 aa linker RFC-25 (fusion standard Freiburg)


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The linker has been order as six separate, EcoRI+AgeI precut primers, which were hybridized in order to obtain the complete linker sequence.

Forward strand (5’->3’) : (89 bp)
aattcgcggccgcttctagatggc|cggcGCGAGCGGCGCGGGCGGCAGCGAAGGCGGCGGCAG|CGAAGGCGGCACCAGCGGCGCGACCa

Reverse strand (5’-3’) : (89 bp)
ccggtGGTCGCGCCG|CTGGTGCCGCCTTCGCTGCCGCCGCCTTCGCTGCC|GCCCGCGCCGCTCGCgccggccatctagaagcggccgcg

Ordered primer:

Forward:         5’aattcgcggccgcttctagatggc 3’

Forward:         5’cggcGCGAGCGGCGCGGGCGGCAGCGAAGGCGGCGGCAG 3’

Forward:         5’CGAAGGCGGCACCAGCGGCGCGACCa 3’

Reverse:          5’ccggtGGTCGCGCCG 3’

Reverse:          5’CTGGTGCCGCCTTCGCTGCCGCCGCCTTCGCTGCC 3’

Reverse:          5’GCCCGCGCCGCTCGCgccggccatctagaagcggccgcg3’


Source

The linker has been order as six separate, EcoRI+AgeI precut primers, which were hybridized in order to obtain the complete linker sequence.

References

Sauer, R. T., T. A. Baker, et al. (2009). "Structures of asymmetric ClpX hexamers reveal nucleotide-dependent motions in a AAA+ protein-unfolding machine." Cell 139(4): 744-56.