Composite

Part:BBa_K3746011

Designed by: Cheung Chi Ming   Group: iGEM21_Hong_Kong_JSS   (2021-10-19)


T7-LacO-DsbAss-FDRA-6His

Description

This is a part design for the Phase I project (2021 iGEM) of Team Hong Kong JSS.

The IPTG inducible secretory expression construct of F420H2-dependent reductase (FDR)-A from Mycobacterium smegmatis (BBa_K3746003). It is the secretory version of expression construct (BBa_K3746010).

The coding sequence of tvLac is driven by T7 promoter (BBa_I719005), a lac operon (BBa_K3076802), strong RBS (BBa_K3076801), a signal peptide for protein secretion (DsbAss (BBa_K3746005)) and a T7 terminator (BBa_K395601).

The FDR-A protein is tagged with a C-terminal 6-His Tag (CATCATCATCATCATCAT) for protein purification.

The DsbAss signal peptide is commonly used in E. coli for protein secretion by signal recognition particle (SRP) pathway.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 189
  • 1000
    COMPATIBLE WITH RFC[1000]
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Categories
Parameters
None