Difference between revisions of "Part:BBa K424017:Design"
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===Design Notes===
 
===Design Notes===
 +
 
We were able to ligate successfully the promotor with the RBS into a plasmid backbone; the reporter with the terminator too; and we get the mutated rhamnosiltransferase 1 gene complex fixed to be compatible with the assembly Standard 10.
 
We were able to ligate successfully the promotor with the RBS into a plasmid backbone; the reporter with the terminator too; and we get the mutated rhamnosiltransferase 1 gene complex fixed to be compatible with the assembly Standard 10.
  
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===References===
 
===References===
 +
 
Assembly Standard Protocol 10
 
Assembly Standard Protocol 10
−
QuikChange Lightning Multi Sit.e-Directed Mutagenesis Kit instruction manual from Stratagene.
+
QuikChange Lightning Multi Site-Directed Mutagenesis Kit Instruction manual, catalog #210515 from Stratagene.

Revision as of 23:37, 27 October 2010

Test plataform for rhamnosyltransferase BioBrick (Rh1AB_BB) expression in E. coli


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 220
    Illegal BamHI site found at 780
    Illegal XhoI site found at 956
    Illegal XhoI site found at 2242
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1135
    Illegal NgoMIV site found at 1856
    Illegal NgoMIV site found at 1969
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 445
    Illegal BsaI site found at 1485
    Illegal BsaI.rc site found at 629
    Illegal BsaI.rc site found at 3135


Design Notes

We were able to ligate successfully the promotor with the RBS into a plasmid backbone; the reporter with the terminator too; and we get the mutated rhamnosiltransferase 1 gene complex fixed to be compatible with the assembly Standard 10.


Source

iGEM, Part registry organization give us the composite parts. The standard Rh1AB part was gene isolated from a Pseudomonas aeruginosa and mutated by a mutagenesis protocol.

References

Assembly Standard Protocol 10 QuikChange Lightning Multi Site-Directed Mutagenesis Kit Instruction manual, catalog #210515 from Stratagene.