Coding
ORF314

Part:BBa_K1846000:Design

Designed by: Luba Prout   Group: iGEM15_Birkbeck   (2015-07-29)

Bacteriophage lambda ORF314



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

To transform ORF-314 sequence into a BioBrick basic part, we had the coding sequence synthesised, including the BioBrick prefix and suffix. The sequence was optimised to remove illegal restriction sites. We restricted both our synthesised ORF-314 and the linearised plasmid backbones (pSB1C3 for shipping, and pSB1K3 for further processing) with EcoRI and PstI restriction enzymes, before ligating the two pieces together using T4 DNA ligase. Success of the cloning procedure was confirmed by restriction with EcoRI and PstI restriction enzymes followed by agarose gel electrophoresis (Figure 1), with pSB1C3 alone and ORF314 alone as controls. The construct was also confirmed by sequencing. ORF-314 was submitted as BBa_K1846000.

BBKiGEM-ORF-314.jpg

Source

The gene fragment was synthesised using sequence from the genome of bacteriophage Lambda.


References