Difference between revisions of "Part:BBa K2457000"

(→‎Usage and Biology)
Line 11: Line 11:
 
<p>Repressor: AraC acts like repressor to both promoters (PBAD e PC) in glucose presence or L-Arabinose absence.</p>
 
<p>Repressor: AraC acts like repressor to both promoters (PBAD e PC) in glucose presence or L-Arabinose absence.</p>
 
<p>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).</p>
 
<p>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).</p>
 +
 +
===Design===
 +
[[File:BBa K2457000 circuit.png]]
 +
 +
Figure 1: BBa_K2457000 Circuit.
  
 
===Characterization===
 
===Characterization===
 
[[File:AraC pBAD electropherogram sequence1.jpeg]]
 
[[File:AraC pBAD electropherogram sequence1.jpeg]]
−
Figure 1:Sequencing electropherogram from BBa_K2457000.  
+
Figure 2:Sequencing electropherogram from BBa_K2457000.  
  
 
[[File:AraC pBAD alignment.png]]
 
[[File:AraC pBAD alignment.png]]
−
Figure 2:Allignment of the designed sequence and our final construction from BBa_K2457000.  
+
Figure 3:Allignment of the designed sequence and our final construction from BBa_K2457000.  
  
 
<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>

Revision as of 09:20, 1 November 2017


araC + pBAD arabinose inducible promoter

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


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

BBa K2457000 circuit.png

Figure 1: BBa_K2457000 Circuit.

Characterization

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

AraC pBAD alignment.png Figure 3:Allignment 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