Difference between revisions of "Part:BBa K3110032:Design"

 
 
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<partinfo>BBa_K3110032 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K3110032 SequenceAndFeatures</partinfo>
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===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 lldP+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-lldP flank and lldD flank-terminator were synthesized by IDT. <br>
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lldP+lldR+lldD were amplified from DH5alpha genome.
  
  
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===References===
  
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===References===
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iGEM ETH zurich 2015<br>
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iGEM NUS Singapore 2016<br>
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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 17:01, 21 October 2019


Strong Promoter Medium RBS lldP+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
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 329
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 3651
    Illegal AgeI site found at 465
    Illegal AgeI site found at 928
    Illegal AgeI site found at 1366
    Illegal AgeI site found at 1525
    Illegal AgeI site found at 1537
    Illegal AgeI site found at 2283
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 3088



Source

Strong promoter-Medium RBS-lldP flank and lldD flank-terminator were synthesized by IDT.
lldP+lldR+lldD were amplified from DH5alpha 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