Coding

Part:BBa_K1321337:Design

Designed by: Chris N Micklem   Group: iGEM14_Imperial   (2014-10-08)
Revision as of 16:52, 12 October 2014 by Xen sm (Talk | contribs) (→‎Design Notes)


sfGFP in Freiburg format (RFC 25)


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
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 10


Design Notes

The BioBrick Part:BBa_I746909 was used as a template for overhang-primer PCR to add the RFC25 prefix and suffix onto the sfGFP coding region. Primers used were as follows:

Forward primer: 5'-GAATTCGCGGCCGCTTCTAGATGGCCGGCcgtaaaggcgaagagctg-3'

Reverse primer: 5'-CTGCAGCGGCCGCTACTAGTATTAACCGGTtttgtacagttcatccataccatg-3'

Capital letters are the RFC25 prefix/suffix overhang, lower-case letters are the portion of the primer initially bound to template.

Source

Made from the existing part Part:BBa_I746909

References

Origin of sfGFP sequence: Pedelacq et al (2006): "Engineering and characterization of a superfolder green fluorescent protein", Nature Biotech 24 (1) January 2006