Composite

Part:BBa_K5040008

Designed by: Chun Ying Yu   Group: iGEM24_CCU-Taiwan   (2024-09-24)


T7promoter-Zif268-rLink-m-hTPH1-PBSII-rlinker-hPCBD1-ZFa-hQDPR-T7terminator

The expression cassette is composed of T7promoter, hTPH1 RBS, Zif268-rLink-m-hTPH1, hPCBD1 RBS, PBSII-rlinker-hPCBD1, hQDPR RBS, ZFa-hQDPR, T7terminator.


Experimental confirmation

After confirming the expression cassette insertion into pST39 vector, we induced the protein expression by treating 0.2mM IPTG at 37°C. To determine whether the expression level increases over time, we conducted inductions for 0, 4, 8 hours.

t-p-q-0-4-8-s.jpeg

Fig. Coomassie Blue-stained SDS-PAGE showing m-hTPH1, hPCBD1, and hQDPR proteins, each fused with Zinc finger proteins, induced with 0.2mM IPTG at 37°C for 0, 4, and 8 hours.


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1362
    Illegal XbaI site found at 39
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1362
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1362
    Illegal BglII site found at 403
    Illegal BamHI site found at 1338
    Illegal XhoI site found at 3086
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1362
    Illegal XbaI site found at 39
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1362
    Illegal XbaI site found at 39
    Illegal NgoMIV site found at 2577
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 20


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