Composite

Part:BBa_K5207012

Designed by: Ruyi Shi   Group: iGEM24_Shanghai-BioX   (2024-09-30)


pPGK1-SmKSL-tADH1

This is an expression cassette consisting of a promoter pPGK1, the SmKSL gene, and a terminator tADH1.


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 1105
    Illegal XhoI site found at 2938
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 3
    Illegal BsaI site found at 2924
    Illegal BsaI.rc site found at 1115
    Illegal BsaI.rc site found at 3174


Usage and Biology

We combined each Level 0 junction module, promoter, CDS, terminator, and ending module, and constructed them into vector pYTK095 through BsaI digestion and ligation, and this step was the construction of Level 1.

Figure 1. Snapgene diagrams of pYTK095

After transformation in E. coli DH10B, we selected the single bacterial colony (pYTK095) that does not fluoresce under UV light from the Petri dishes. It was used for colony PCR amplification and validation.

Figure 2. Fluorescent labeling screening of pYTK095

Figure 3. Validation plot of pYTK095 gel electrophoresis

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