Difference between revisions of "Part:BBa K4800054"

 
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<partinfo>BBa_K4800054 short</partinfo>
 
<partinfo>BBa_K4800054 short</partinfo>
  
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davB-davA-GabT gene encodes for carboxylate reductase, alcohol dehydrogenase and transaminase. These enzymes can realize the metabolic pathway from lysine to glutarate semialdehyde, and the three enzymes form a synthetic module including 5-HV.
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We use the laboratory conserved plasmid pET22b-T7-davB-T7-davAas templates, the fragments of davB and davA were amplified by PCR with a consensus RBS and Ptrc respectively. Subsequently, the Ptrc-RBS-davB-Ptrc-RBS-davA fragment was obtained by overlap PCR, which was then ligated into the linearized vector pTrc99a-GabT by In-fusion cloning method to obtain plasmid pTtrc99a-davB-davA-GabT.
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<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Revision as of 12:09, 11 October 2023


pTrc99a-davB-davA-GabT

We use the laboratory conserved plasmid pET22b-T7-davB-T7-davAas templates, the fragments of davB and davA were amplified by PCR with a consensus RBS and Ptrc respectively. Subsequently, the Ptrc-RBS-davB-Ptrc-RBS-davA fragment was obtained by overlap PCR, which was then ligated into the linearized vector pTrc99a-GabT by In-fusion cloning method to obtain plasmid pTtrc99a-davB-davA-GabT.


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 5122
    Illegal XbaI site found at 6489
    Illegal PstI site found at 6501
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 5122
    Illegal PstI site found at 6501
    Illegal NotI site found at 2808
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 5122
    Illegal BglII site found at 3897
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 5122
    Illegal XbaI site found at 6489
    Illegal PstI site found at 6501
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 5122
    Illegal XbaI site found at 6489
    Illegal PstI site found at 6501
    Illegal NgoMIV site found at 4057
    Illegal AgeI site found at 3464
    Illegal AgeI site found at 3713
    Illegal AgeI site found at 5535
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 6685
    Illegal BsaI site found at 7753
    Illegal SapI.rc site found at 767