Regulatory

Part:BBa_K2457000

Designed by: Maria Astolfi, Giovanna Maklouf, Juliana Vitoriano   Group: iGEM17_Amazonas_Brazil   (2017-07-08)
Revision as of 17:00, 1 November 2017 by Giomaklouf (Talk | contribs)


araC + pBAD arabinose inducible promoter

Arabinose inducible promoter pBAD forward + araC regulatory reverse gene under control of pC promoter and tryptophan terminator sequence.

BBa K2457000 circuit.png

Figure 1: BBa_K2457000 Circuit.


Usage and Biology

Inductor: L-Arabinose

Repressor: AraC acts like repressor to both promoters (PBAD e PC) in glucose presence or L-Arabinose absence.

The BBa_K2457000 includes approximately 1298 nucleotides and is used to regule transgene expression in Escherichia coli bacterias. This construction is composed by the araBAD (PBAD) promoter, which produces medium force, the L-Arabinose operon, in 5’ → 3’ direction with a upstream native RBS, two regulatory sites: I1 and I2, and two operators sites AraO, O1 and O2. It is composed also by the promoter PC that transcribes the regulatory protein AraC gene, followed to tryptophan transcription terminator, in 3’ → 5’ (reverse) (KUHLMAN & COX, 2010).

Design

Characterization

AraC pBAD electropherogram sequence1.jpeg Figure 2:Sequencing electropherogram from BBa_K2457000.

AraC pBAD alignment.png Figure 3:Alignment of the designed sequence and our final construction from BBa_K2457000.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1207
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 1042
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 1024


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Categories
Parameters
None