Difference between revisions of "Part:BBa K4137006"
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===Usage and Biology===
 
===Usage and Biology===
  
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There is an illegal site (PstI) in the original sequence of CrtE (see Figure 1a). In order to remove the illegal site, we designed two primers matching with the region but change the PstI site (CTGCAG) into CTGCAA (see Figure 1b). Then we amplified CrtE into two parts (see Figure 1c, 1d) and then linked them again using overlap PCR (see Figure 1e). In this way we removed the illegal site but did not change the amino sequence. But we finished this job too late, leading us no time to cloned our engineered CrtE into the plasmid pACYC184-M, which was a pity.
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[[File:pmles-ccda-his.png.png|800px|thumb|center|Fig.1 Malate-inducible ccdA full construct.]]
  
 
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Revision as of 03:24, 9 October 2022


CcdA expressing construct

The mleR regulatory site which comes from p_mles binds with the RBS and CcdA, the antitoxin that will bind with CcdB when induced by malate acid and detoxifies it, the purification tag 6xHis, and the terminator B1006 (BBa_B1006). This sequence codes for CcdA, the antitoxin which assures chi18h8’s production around the plant roots.

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]