Difference between revisions of "Part:BBa K4137006"

 
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===Construct Designs===
 
===Construct Designs===
  
We attached a 6x His-Tag downstream of the ccdA sequence for purification purposes. A p_mleS promoter, which induces protein production when malate is present, as well as RBS, are attached upstream to the side of the open reading frame (ccdA-6x his tag). The terminator BBa_B1006 is attached downstream of the sequence.  
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We attached a 6x His-Tag downstream of the ccdA sequence to determine the basal production of the antitoxin. A p_mleS promoter, which induces protein production when malate is present, as well as a 5'UTR RBS (AAGGAG), are attached upstream to the side of the open reading frame (ccdA-6x his tag). The terminator BBa_B1006 is attached downstream of the sequence.  
  
===Characterization===
 
  
  
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===Sequence and Features===
 
===Sequence and Features===
 
<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>

Latest revision as of 07:52, 10 October 2022


CcdA expressing construct

This sequence codes for CcdA, the antitoxin which assures chi18h8’s production around the plant roots. The mleR regulatory site comes from p_mles, which binds with the RBS and CcdA, the antitoxin that will bind with CcdB when induced by malate acid. This will detoxify ccdB. The purification tag 6xHis, and the terminator B1006 (BBa_B1006) are flanked on the end of the open reading frame.

Fig.1 Construct design for ccdA antitoxin secretion.

Construct Designs

We attached a 6x His-Tag downstream of the ccdA sequence to determine the basal production of the antitoxin. A p_mleS promoter, which induces protein production when malate is present, as well as a 5'UTR RBS (AAGGAG), are attached upstream to the side of the open reading frame (ccdA-6x his tag). The terminator BBa_B1006 is attached downstream of the sequence.


Sequence and Features

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]