Difference between revisions of "Part:BBa K1783003:Design"

 
 
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===Design Notes===
 
===Design Notes===
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PCR of Hok-Sok component was difficult due to high inherent secondary structure of the gene sequences. DMSO was needed in order to break this structure.
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Construct was assembled using Gibson assembly of a gBlock of K1783001 (Hok/Sok) and K1783002 (Const. Unstable RFP) into a pSB1C3 backbone.
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PCR amplification for primer extension of Hok/Sok component was difficult due to high inherent secondary structure of the gene sequences. Therefore, the Gibson Assembly of the Hok/Sok and Const. Unstable RFP proved to be difficult.
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===Source===
 
===Source===
  
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See construction parts.
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Hok/Sok from K1783001 and constitutive unstable RFP from K1783002.
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Hok/Sok is from the E.coli R1 plasmid. RFP is from coral. The promoters are from the E.coli genome. LVA-tag described by Andersten et al.  
  
 
===References===
 
===References===
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1998, www.ncbi.nlm.nih.gov/pmc/articles/PMC106306/

Latest revision as of 15:09, 26 September 2015

Unstable RFP under Hok/Sok Regulation


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 587
    Illegal NheI site found at 610
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 1199
    Illegal AgeI site found at 1311
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Construct was assembled using Gibson assembly of a gBlock of K1783001 (Hok/Sok) and K1783002 (Const. Unstable RFP) into a pSB1C3 backbone. PCR amplification for primer extension of Hok/Sok component was difficult due to high inherent secondary structure of the gene sequences. Therefore, the Gibson Assembly of the Hok/Sok and Const. Unstable RFP proved to be difficult.

Source

Hok/Sok from K1783001 and constitutive unstable RFP from K1783002. Hok/Sok is from the E.coli R1 plasmid. RFP is from coral. The promoters are from the E.coli genome. LVA-tag described by Andersten et al.

References

1998, www.ncbi.nlm.nih.gov/pmc/articles/PMC106306/