Difference between revisions of "Part:BBa K2043005:Design"
Line 7: Line 7:
  
 
===Design Notes===
 
===Design Notes===
−
His-tag added for enhanced purification. <br>
+
Fabric Binding Domain 1 was fused downstream in the 5' end of the original <i>catA</i> CDS <br>
−
Gene was codon optimised for expression in <i>E. coli</i>
+
A linker was introduced in the DNA sequence between FBD1 and the CDS of <i>catA</i> <br>
−
 
+
His-tag is added in the C-terminal for protein purification <br>
−
 
+
The sequence was codon optimized for <i>E. coli</i> and restriction sites for the restriction enzymes BpiI, BsaI and BsmBI were avoided. <br>
 +
The sequence was confirmed by sequencing and no mutations were observed. <br>
 +
The construction was done by Golden Gate technique using BsaI as restriction enzyme to obtain the desired cohesive ends.
  
 
===Source===
 
===Source===

Revision as of 15:50, 27 October 2016


xylE-FBD1 from Pseudomonas putida codon optimized for E. coli


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 700
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Fabric Binding Domain 1 was fused downstream in the 5' end of the original catA CDS
A linker was introduced in the DNA sequence between FBD1 and the CDS of catA
His-tag is added in the C-terminal for protein purification
The sequence was codon optimized for E. coli and restriction sites for the restriction enzymes BpiI, BsaI and BsmBI were avoided.
The sequence was confirmed by sequencing and no mutations were observed.
The construction was done by Golden Gate technique using BsaI as restriction enzyme to obtain the desired cohesive ends.

Source

Pseudomonas putida

References