Composite

Part:BBa_K3875006

Designed by: Yimiao Lin   Group: iGEM21_BUCT   (2021-09-09)
Revision as of 09:12, 18 October 2021 by Ekko (Talk | contribs)


Usage and Biology

Design: Contained gene: trpE ( BBa_K3875009), trpG ( BBa_K3875003), trpC ( BBa_K3875002).
This part could express two enzymes, indole-3-glycerol-phosphate synthase (IGPS, encoded by trpC) and anthranilate synthase (AS, encoded by trpE).
Erythritos-4-phosphate (E4P) from the pentose phosphate pathway (PPP) is condensed with phosphoenolpyruvate (PEP) from the glycolysis pathway (EMP) to form 3-deoxy-D-arabinheptanose-7-phosphate (DAHP).
After 7 steps, DAHP is converted to chorismate. Our trpE and trpC genes in E. coli promote the chorismate into the latter half of the 5-HTP synthesis pathway (Figure 1).





Method: The pcd and p4h genes were amplified by PCR, then the two genes and pSA74 were digested by restriction endonuclease, and finally pSA74-lac-trpE-trpG-AntrpC-T1 was obtained by ligase T4. Then HindIII and BamHI were used for digestion verification.
Result: The result of agarose gel electrophoresis proves that our plasmid has been successfully constructed .

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 600
    Illegal BglII site found at 661
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 4128
  • 1000
    COMPATIBLE WITH RFC[1000]


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Parameters
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