P<sub>C</sub> is a constitutive promoter that expresses the downstream <i>araC</i> gene. P<sub>BAD</sub> promoter is an inducible promoter, P<sub>BAD</sub> contains sequences of P<sub>C</sub>. It was verified by iGEM 2017 Amazonas_Brazil team that the induction effect of arabinose could be realized by using only <i>araC</i> and P<sub>BAD</sub>.<br><br>
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===Usage and Biology===
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Arabinose operon is composed of AraC protein, P<sub>BAD</sub> and P<sub>C</sub>. The <i>araC</i> gene translates from the P<sub>C</sub> to the left.<br>
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We used the P<sub>C</sub> promoter to express AraC constitutively, and constituted the sensing part of the arabinose sensing system.<br>
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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.<br>
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The L-Arabinose operon is naturally present in <i>E. coli</i>.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 P<sub>BAD</sub> 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.
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We connected the target genes downstream of P<sub>BAD</sub> to constitute the effect part of arabinose sensing system, and constructed the whole arabinose sensing system on the same plasmid with P<sub>C</sub> and <i>araC</i>.<br>
Arabinose induces green fluorescent protein expression: (链接a01+c09)<br><br>
==<b>Design</b>==
==<b>Design</b>==
Revision as of 11:06, 22 October 2020
Arabinose production
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
