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Php4d

Part:BBa_K1592004

Designed by: Shuyan Tang   Group: iGEM15_HUST-China   (2015-09-01)

promoter hp4d

The recombinant promotor hp4d, nearly unaffected by pH, carbon source, nitrogen source and some conditions, can strongly promote gene expression in any culture medium(Madzak et al., 2000). Promoter hp4d is composed of four tandem repeated UAS1 sequences in upstream and the small LEU2 promoter, which is the sequence between TATA box and start condon ATG of LEU2 promoter in downstream. UAS1 is the upsteam activating sequence of the pXPR2, which is less affected by other conditions. Though any inducers are non-essential for activiation of php4d during cultivating, hp4d, activated by unknown element, isn't constitutive promoter. The gene promoted by promoter hp4d usually expresses at the early stage of stabilization, called growth phase dependent (Nicaud et al., 2002). This feature, separating the strain growth and producing foreign protein, is useful for production of foreign protein, which avoids the harm by expressing foreign protein.


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
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 92
    Illegal BsaI.rc site found at 201
    Illegal BsaI.rc site found at 310
    Illegal BsaI.rc site found at 419

Usage

We use Php4d and construct circuits (Figure 1 and Figure 2) to express the cell surface display Si-tag system of Yarrowia Lipolytica (more experimental details see BBa_K1592007- BBa_K1592014) and the LIP2 prepro-Mcfp3(BBa_K1592003) for validating the parts’ function.

Figure1: Php4d+LIP2 prepro-Mcfp+terminator
Figure2: Php4d+LIP2 prepro- Si-tag -YLcwp3 +terminator



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