Composite

Part:BBa_K3447102

Designed by: Fei Cao, Yu Ma   Group: iGEM20_Jilin_China   (2020-10-22)
Revision as of 11:08, 22 October 2020 by Alswey (Talk | contribs)


Arabinose production

PC is a constitutive promoter that expresses the downstream araC gene. PBAD promoter is an inducible promoter, PBAD contains sequences of PC. It was verified by iGEM 2017 Amazonas_Brazil team that the induction effect of arabinose could be realized by using only araC and PBAD.

Usage and Biology

Arabinose operon is composed of AraC protein, PBAD and PC. The araC gene translates from the PC to the left.
We used the PC promoter to express AraC constitutively, and constituted the sensing part of the arabinose sensing system.
In the arabinose operon, it is regulated by the AraC operon and activates the expression of downstream arabinose metabolism genes in the presence of arabinose inducers.
The L-Arabinose operon is naturally present in E. coli.The regulatory protein AraC ACTS as a dimer. The operon contains two cis-type control elements: The PC promoter for the synthesis of AraC and the PBAD for the synthesis of the enzyme needed for the catabolism of L-Arabinose. PBAD contains three half sites, each of which binds to a subunit of ARAC-O2, I1, and I2. The O1 site is composed of O1L and O1R half sites that bind two subunits. Half of O2 is in the araC coding region. We connected the target genes downstream of PBAD to constitute the effect part of arabinose sensing system, and constructed the whole arabinose sensing system on the same plasmid with PC and araC.
Arabinose induces endotoxin expression: (链接a01+c07)
Arabinose induces green fluorescent protein expression: (链接a01+c09)

Design

Design Notes

We added some synonymous mutations to avoid part rules.


Source

We found this sequence data in the previous iGEM teams (Glasgow 2017) and in GenBank.

References

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 22
    Illegal NheI site found at 45
    Illegal NheI site found at 166
    Illegal NheI site found at 891
    Illegal NheI site found at 914
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1
    Illegal BsaI.rc site found at 76
    Illegal BsaI.rc site found at 570
    Illegal BsaI.rc site found at 1657
    Illegal BsaI.rc site found at 1817
    Illegal SapI.rc site found at 624


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