Difference between revisions of "Part:BBa K3110033:Design"

 
 
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<partinfo>BBa_K3110033 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K3110033 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 -Weak 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+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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lldP+lldR+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 17:06, 21 October 2019


Strong Promoter Weak 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 327
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 3649
    Illegal AgeI site found at 463
    Illegal AgeI site found at 926
    Illegal AgeI site found at 1364
    Illegal AgeI site found at 1523
    Illegal AgeI site found at 1535
    Illegal AgeI site found at 2281
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 3086

Source

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