Plasmid

Part:BBa_K4289016

Designed by: Kong Yangyang   Group: iGEM22_Nanjing_HS   (2022-09-29)


pGEX-GST-GK

pGEX-GST-GK

Contribution

pGEX-4T-1-backbone is an E. coli expression vector, which employs tac promoter to regulate the expression of exogenous genes. In order to increase protein solubility, we inserted the hGK2 DNA fragment into the BamHI and SalI sites, and the GST tag is fused at the N-terminal of the protein, and the tag could be removed by thrombin. The vector is a high-copy-number plasmid. When expressed in the prokaryotic system, the Amp+ resistance can be used to screen the right colony. These plasmid backbones we submitted could be used to express different proteins in the future.

Figure 1. The map of plasmid-backbone pGEX-4T-1.







Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 970
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 970
    Illegal NotI site found at 11
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 4934
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 970
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 970
  • 1000
    COMPATIBLE WITH RFC[1000]


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Parameters
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