Coding
stf

Part:BBa_K1846004:Design

Designed by: Luba Prout   Group: iGEM15_Birkbeck   (2015-09-17)

stf (short tail fibre) gene of bacteriophage lambda


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
    COMPATIBLE 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 SpeI restriction enzymes followed by agarose gel electrophoresis (Figure 1) and in part by Sanger sequencing. This BioBrick was registered as part BBa_K1846004.

BBK-stf-gene.jpg


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.


References