Difference between revisions of "Part:BBa K191004"
Line 5: Line 5:
  
 
Our first read-out system consists of the Trp promoter followed by the RFP gene. In E. coli, the Trp promoter is situated in front of the Trp operon, which contains the genes necessary for the tryptophane biosynthetic pathway. In brief, genes placed after the Trp promoter should be repressed in the presence of tryptophane.  
 
Our first read-out system consists of the Trp promoter followed by the RFP gene. In E. coli, the Trp promoter is situated in front of the Trp operon, which contains the genes necessary for the tryptophane biosynthetic pathway. In brief, genes placed after the Trp promoter should be repressed in the presence of tryptophane.  
 +
 +
<font size="3">'''Read-Out n°1 BioBrick'''</font>
 +
 +
This BioBrick was synthesized using the protocol we developed with the Klenow fragment. To do this, two long primers which together contained the sequence for the Trp promoter were ordered; these primers could self-anneal in the reaction mix, and the second strands that were missing at the extremities of the primers were fully synthesized using the Klenow fragment, instead of a classic extension using the TAQ DNA Polymerase.
 +
 +
For more information, here is the [http://2009.igem.org/wiki/index.php?title=Team:EPF-Lausanne/Protocols/Klenow complete protocol].
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Revision as of 15:49, 21 October 2009

TRP promoter - RBS - RFP - Term

Readout 1, RFP's transcription controlled by TRP operon

Our first read-out system consists of the Trp promoter followed by the RFP gene. In E. coli, the Trp promoter is situated in front of the Trp operon, which contains the genes necessary for the tryptophane biosynthetic pathway. In brief, genes placed after the Trp promoter should be repressed in the presence of tryptophane.

Read-Out n°1 BioBrick

This BioBrick was synthesized using the protocol we developed with the Klenow fragment. To do this, two long primers which together contained the sequence for the Trp promoter were ordered; these primers could self-anneal in the reaction mix, and the second strands that were missing at the extremities of the primers were fully synthesized using the Klenow fragment, instead of a classic extension using the TAQ DNA Polymerase.

For more information, here is the [http://2009.igem.org/wiki/index.php?title=Team:EPF-Lausanne/Protocols/Klenow complete protocol].

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal SpeI site found at 43
    Illegal SpeI site found at 51
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal SpeI site found at 43
    Illegal SpeI site found at 51
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal SpeI site found at 43
    Illegal SpeI site found at 51
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal SpeI site found at 43
    Illegal SpeI site found at 51
    Illegal AgeI site found at 666
    Illegal AgeI site found at 778
  • 1000
    COMPATIBLE WITH RFC[1000]