Difference between revisions of "Part:BBa K1846004:Design"
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Revision as of 22:59, 20 September 2015
stf (short tail fibre) gene of bacteriophage lambda
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
This gene has been assembled using the synthesised ORF-401 with a linker sequence (removing the frameshift mutation) and ORF-314, also synthesised (available separately as BioBrick BBa_K1846000). The sequences were optimised to remove illegal restriction sites to make the sequence BioBrick-compatible. We cloned the two open reading frames and a linker into a single in-frame coding sequence (into a pSB1C3 backbone) using Gibson assembly. Success of the cloning procedure was confirmed by restriction with EcoRI and PstI restriction enzymes, and separately with EcoRV, followed by DNA agarose electrophoresis (Figure 1). This BioBrick was registered as part BBa_K1846004.
Source
The gene fragment was synthesised using sequence from the genome of bacteriophage Lambda adding 1 bp just after ORF-401 to remove the frameshift mutation.