Regulatory

Part:BBa_K3726079

Designed by: Santiago Barragán Ariza   Group: iGEM21_MADRID_UCM   (2021-10-08)
Revision as of 23:06, 21 October 2021 by Sbarraga (Talk | contribs)

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P_PpsbA*-RiboJ

This part is a strong constitutive promoter for cyanobacteria. It is a modified version of PpsbA* promoter originally documented by Wang et al. (B. Wang, C. Eckert, P. Maness and J. Yu, "A Genetic Toolbox for Modulating the Expression of Heterologous Genes in the Cyanobacterium Synechocystis sp. PCC 6803", 2021.)

The PpsbA-RiboJ includes an insulator sequence after the promoter region. The RiboJ insulator (BBa_K1679038) has been included. This incorporation helps buffering the genetic context effect of the upstream promoter sequence in the design of synthetic circuits. Likewise, the addition of RiboJ has shown to enhance expression levels, becoming this promoter stronger than PpsbA* itself. ____ PpsbA* is an hybrid promoter where the strong light-inducible PpsbA promoter from Amaranthus hybridus and the Cu2+ inducible promoter petE from Synechocystis PCC6803 has been combined.

The PpsbA-RiboJ includes an insulator sequence after the promoter region. The RiboJ insulator (BBa_K1679038) has been included. This incorporation helps buffering the genetic context effect of the upstream promoter sequence in the design of synthetic circuits. Likewise, the addition of RiboJ has shown to enhance expression levels, becoming this promoter stronger than PpsbA* itself.

The part is a phytobrick compatible MoClo Lv.0 part, assembled in the universal phytobrick entry vector BBa_K2560001.


Contribution from MADRID_UCM

Group: MADRID_UCM (4C_Fuels Project) Author: Jorge Fernández, Alvaro Luego, Santiago Barragán Ariza, Lucia Tapia Peña. Summary: Promoter PpsbA-RiboJ have demonstrated one of the highest expression levels in Synechococcus elongatus genre as well as related cyanobacteria species. This promoter element has been adapted to Phytobricks standard and succesfully assembled.

This phytobrick compatible promoter is part of a wider collection of regulatory elements for cyanobacteria, which are part of MADRID_UCM 2021 team contribution to iGEM community.


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 234
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 234
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 234
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 234
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 234
  • 1000
    COMPATIBLE WITH RFC[1000]


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