Difference between revisions of "Part:BBa K1846004:Design"
 
(One intermediate revision by the same user not shown)
Line 7: Line 7:
 
===Design Notes===
 
===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 [https://parts.igem.org/Part:BBa_K1846000 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 followed by DNA agarose electrophoresis (Figure 1) and in part by Sanger sequencing. This BioBrick was registered as part [https://parts.igem.org/Part:BBa_K1846004 BBa_K1846004].
+
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 [https://parts.igem.org/Part:BBa_K1846000 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 [https://parts.igem.org/Part:BBa_K1846004 BBa_K1846004].
  
 
[[File:BBK-stf-gene.jpg|350px|thumb|none|]]
 
[[File:BBK-stf-gene.jpg|350px|thumb|none|]]

Latest revision as of 00:22, 21 September 2015

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