Difference between revisions of "Part:BBa K3110010:Design"

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===Design Notes===
 
===Design Notes===
 
lldD was codon optimized to meet synthesis requirements and to remove the illegal EcoRI site present in the genomic lldD.
 
lldD was codon optimized to meet synthesis requirements and to remove the illegal EcoRI site present in the genomic lldD.
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−
 
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===Source===
 
===Source===
  
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lldD was Synthesized from Twist Bioscience and was joined to Promoter+RBS and Terminator using SOEing (Spliced Overlap Extension) PCR.
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The lldD Synthesized from Twist Bioscience. Specific Promoter and RBS with D flank and Terminator with D flank was ordered from IDT and all the 3 parts were merged together using SOEing (Spliced Extension Overlapped) PCR.  
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===References===
 
===References===
 
Aguilera, L., Campos, E., Gimenez, R., Badia, J., Aguilar, J., & Baldoma, L. (2008). Dual Role of LldR in Regulation of the lldPRD Operon, Involved in L-Lactate Metabolism in Escherichia coli. Journal of Bacteriology, 190(8), 2997–3005. doi:10.1128/jb.02013-07
 
Aguilera, L., Campos, E., Gimenez, R., Badia, J., Aguilar, J., & Baldoma, L. (2008). Dual Role of LldR in Regulation of the lldPRD Operon, Involved in L-Lactate Metabolism in Escherichia coli. Journal of Bacteriology, 190(8), 2997–3005. doi:10.1128/jb.02013-07

Latest revision as of 20:58, 21 October 2019


Weak Promoter Strong RBS lldD


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1202
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 639


Design Notes

lldD was codon optimized to meet synthesis requirements and to remove the illegal EcoRI site present in the genomic lldD.


Source

The lldD Synthesized from Twist Bioscience. Specific Promoter and RBS with D flank and Terminator with D flank was ordered from IDT and all the 3 parts were merged together using SOEing (Spliced Extension Overlapped) PCR.

References

Aguilera, L., Campos, E., Gimenez, R., Badia, J., Aguilar, J., & Baldoma, L. (2008). Dual Role of LldR in Regulation of the lldPRD Operon, Involved in L-Lactate Metabolism in Escherichia coli. Journal of Bacteriology, 190(8), 2997–3005. doi:10.1128/jb.02013-07