Difference between revisions of "Part:BBa K1321337:Design"

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(→‎Design Notes)
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===Design Notes===
 
===Design Notes===
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OEPCR
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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===
 
===Source===

Revision as of 16:52, 12 October 2014


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