Difference between revisions of "Part:BBa K3110022:Design"

 
 
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<partinfo>BBa_K3110022 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K3110022 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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Strong promoter -Strong RBS-lldP flank and lldD flank-terminator were synthesized by IDT. <br>
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lldD was Synthesized from Twist Bioscience and was joined to lldP 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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lldP+lldD was amplified from DH5alpha genome.
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===References===
 
===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 17:57, 21 October 2019


Strong Promoter Strong RBS lldP+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 328
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 2864
    Illegal AgeI site found at 464
    Illegal AgeI site found at 927
    Illegal AgeI site found at 1365
    Illegal AgeI site found at 1524
    Illegal AgeI site found at 1536
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 2301

Source

Strong promoter -Strong RBS-lldP flank and lldD flank-terminator were synthesized by IDT.
lldP+lldD was 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