Difference between revisions of "Part:BBa K2739008:Design"
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The length of the Bktb is 1185bp and excess the 1kb length limit of the IDT sponsor limitation to be generated. Therefore two overlapped fragments of Bktb were then designed and generated by IDT. A set of PCR primer were then design to generate the whole Bktb through the PCR amplification.   
 
The length of the Bktb is 1185bp and excess the 1kb length limit of the IDT sponsor limitation to be generated. Therefore two overlapped fragments of Bktb were then designed and generated by IDT. A set of PCR primer were then design to generate the whole Bktb through the PCR amplification.   
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https://static.igem.org/mediawiki/2018/1/1c/T--Edinburgh_OG--Notebook_-_bktb_1.png
  
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Figure 1. B1 and B2 fragments were codon-optimised for E. coli and synthesised by IDT. The 696 bp overlap was in green and restriction enzyme sites were coloured by red. Two pairs of primers were ordered from IDT to amplify bktB, among which B1 forward primer and B2 reverse primer were used to amplify bktB fragments.
  
  

Revision as of 20:27, 16 October 2018


Bktb


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
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 831


Design Notes

The length of the Bktb is 1185bp and excess the 1kb length limit of the IDT sponsor limitation to be generated. Therefore two overlapped fragments of Bktb were then designed and generated by IDT. A set of PCR primer were then design to generate the whole Bktb through the PCR amplification. T--Edinburgh_OG--Notebook_-_bktb_1.png

Figure 1. B1 and B2 fragments were codon-optimised for E. coli and synthesised by IDT. The 696 bp overlap was in green and restriction enzyme sites were coloured by red. Two pairs of primers were ordered from IDT to amplify bktB, among which B1 forward primer and B2 reverse primer were used to amplify bktB fragments.


Source

The sequence of present Bktb is isolated from Ralstonia Eutropha and codon optimised to E.coli. The original sequence would be accessed in NCBI: NC_015726.1.

The sequence is codon optimised by IDT.

References

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