Difference between revisions of "Part:BBa K1321337:Design"
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===Design Notes=== | ===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=== | ===Source=== |
Revision as of 16:52, 12 October 2014
sfGFP in Freiburg format (RFC 25)
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE 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