Composite

Part:BBa_K3746007

Designed by: Cheung Chi Ming   Group: iGEM21_Hong_Kong_JSS   (2021-10-18)


T7-LacO-DsbAss-tvLac-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 laccase from Trametes versicolor (tvLac) (BBa_K3746002). It is the secretory version of expression construct (BBa_K3746006).

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 laccase protein is tagged with a 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
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 135
    Illegal PstI site found at 1245
    Illegal PstI site found at 1288
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 135
    Illegal PstI site found at 1245
    Illegal PstI site found at 1288
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1643
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 135
    Illegal PstI site found at 1245
    Illegal PstI site found at 1288
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 135
    Illegal PstI site found at 1245
    Illegal PstI site found at 1288
    Illegal AgeI site found at 227
    Illegal AgeI site found at 1040
    Illegal AgeI site found at 1470
  • 1000
    COMPATIBLE WITH RFC[1000]
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Categories
Parameters
None