Difference between revisions of "Part:BBa K2442397"
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We designed a PCR reaction to generate a C-terminal GFP2 part from [https://parts.igem.org/Part:BBa_E0040 E0040] which would work alongside the existing N-terminal GFP1 part [https://parts.igem.org/Part:BBa_K1789003 K1789003] made by NUDT_CHINA 2015. Primers were designed to amplify the coding sequence of the 83 aa C-terminal fragment as a biobrick compatible part. | We designed a PCR reaction to generate a C-terminal GFP2 part from [https://parts.igem.org/Part:BBa_E0040 E0040] which would work alongside the existing N-terminal GFP1 part [https://parts.igem.org/Part:BBa_K1789003 K1789003] made by NUDT_CHINA 2015. Primers were designed to amplify the coding sequence of the 83 aa C-terminal fragment as a biobrick compatible part. | ||
− | [[Image: | + | [[Image:Glasgow_ANDgate_Table_3.png|450px|thumb|center|'''Figure 1:''' Split GFP C-terminal (GFP2) amplification primers. All sequences represented 5’-3’. Colours denote features: blue text = flanking DNA; yellow highlight = Biobrick prefix (F) or suffix (R); purple = new start codon; unformatted text = annealing section of primer. Underlined sequence denotes an error that is discussed in the results section.]] |
Revision as of 02:47, 2 November 2017
GFP C-term split subunit (GFP2)
Design
We designed a PCR reaction to generate a C-terminal GFP2 part from E0040 which would work alongside the existing N-terminal GFP1 part K1789003 made by NUDT_CHINA 2015. Primers were designed to amplify the coding sequence of the 83 aa C-terminal fragment as a biobrick compatible part.
Sequencing of the PCR for the GFP C-terminal part revealed an error – the PCR design had incorporated an incorrect version of the BioBrick prefix.
Usage and Biology
Sequence and Features
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 BsaI.rc site found at 176