Difference between revisions of "Part:BBa K4137006"

Revision as of 06:32, 9 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 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.

Characterization

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]

@@ Line 9: / Line 9: @@
 ===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, is attached upstream to the side of the open reading frame + RBS. The terminator BBa_B1006 is attached downstream of the sequence.
+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.
 ===Characterization===