Composite

Part:BBa_K4836012

Designed by: A Adithya Hitesh, M V P S Sriyanth, Sanchi Chaware, G Manonmani, Dhruvi Panchal, Akhshaya Mohan   Group: iGEM23_IISER-Tirupati-India   (2023-10-10)


SNAT and COMT expression cassette with Fluorescent reporter proteins

This part consists of Serotonin N-Acetyl Transferase(SNAT)+TagBFP fusion and Caffeic acid O-Methyl Transferase(COMT)+mCherry fusion under T7 IPTG induced promoter. This cassette is designed to check for the expression of SNAT and COMT proteins.

We planned to check the expression of SNAT and COMT with the production of TagBFP and mCherry respectively. We cloned our genetic circuit into pJUMP by Golden Gate assembly. The size of the cloned plasmid is 5833bp. We performed colony PCR to check the positive colony. The expected sizes of amplicons are Plasmid forward and insert reverse 1248bp Insert forward and Plasmid Reverse 1063bp

We performed restriction digestion with HindIII and EcoRV single cutter enzymes to further confirm the colony, both enzyme sites were present in the insert at different location. The expected band size is 5833bp.
Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 3226
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
None