Difference between revisions of "Part:BBa K3110035:Design"

 
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<partinfo>BBa_K3110035 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K3110035 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+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 the 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
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===Characterization===
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Due to shortage of time we could not characterize this construct.

Revision as of 17:22, 21 October 2019


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

Characterization

Due to shortage of time we could not characterize this construct.