Coding

Part:BBa_K5013001

Designed by: Zhang Yuxi   Group: iGEM23_Squirrel-CHN   (2023-10-11)
Revision as of 05:08, 11 October 2023 by Xiaoxi (Talk | contribs)

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TP901 recognizes

This sequence contains the TP901 recognition sites attB (5' terminal) and attP (3' terminal). Where the Target sequence is replaced with the reverse T7 promoter. When TP901 recognizes this sequence, it turns over the T7 promoter to initiate downstream gene expression. In order to avoid mutual interference between the 3' terminal region of the promoter and the 5' terminal of the ORF, we added an insulator RiboJ downstream of the attP recognition site.

Usage and Biology

Figure 1 The principle of TP901 integrase flip the target DNA fragment.

We have designed a sophisticated system for in vitro degradation of phenylalanine for adjuvant therapy in patients with phenylketonuria. We introduce a trans-cinnamic acid (TCA) promoter (pSenCA) upstream of the TP901 promoter. When the phenylalanine degradation product TCA is detected, pSenCA initiates the translation of TP901 integrase. TP901 integrase recognizes the attB/attP site, makes the T7 promoter reverse face the PAL gene sequence, initiates the PAL translation, and the strong T7 promoter rapidly produces a large amount of PAL enzyme without inhibiting the growth of bacteria, thus promoting the degradation of phenylalanine.

Figure 2 TCA inducible promoter-controlled integrase system.



Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 92
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 92
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 92
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 73


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