Difference between revisions of "Part:BBa K3110019:Design"

 
 
Line 7: Line 7:
  
 
===Design Notes===
 
===Design Notes===
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lldD was codon optimized to meet synthesis requirements and to remove the illegal EcoRI site present in the genomic lldD.
 
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===Source===
 
===Source===
  
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lldD was Synthesized from Twist Bioscience and was joined to lldR using SOEing (Spliced Overlap Extension) PCR. This was further joined to Promoter+RBS and Terminator using SOEing (Spliced Overlap Extension) PCR.
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Strong promoter -Medium RBS-lldR flank and lldD flank-terminator were synthesized by IDT. <br>
 +
lldR+lldD was amplified from the genome.
 +
 
  
  
  
 
===References===
 
===References===
 +
iGEM ETH zurich 2015<br>
 +
iGEM NUS Singapore 2016<br>
 +
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 15:39, 21 October 2019


Weak Promoter Strong RBS lldR+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 1988
    Illegal AgeI site found at 620
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1425


Design Notes

Source

Strong promoter -Medium RBS-lldR flank and lldD flank-terminator were synthesized by IDT.
lldR+lldD was amplified from the genome.



References

iGEM ETH zurich 2015
iGEM NUS Singapore 2016
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