Regulatory

Part:BBa_K3122003

Designed by: Laura Sanchez Lara   Group: iGEM19_MADRID_UCM   (2019-10-16)
Revision as of 02:48, 22 October 2019 by Laulara (Talk | contribs)


Constitutive promoter J23107 (TypeIIS adapted)

This part is the constitutive medium promoter, from Anderson family, was adapted to type IIS enzymes standard.

It has been used to express our Level 1 constructs related to membrane proteins. The reason behind this choice relays on its relative strength among other members of the family: it is a medium- strength regulator (0.36) which should not express too much membrane protein as to induce cell lysis.

Characterization

Characterization of this part has been conducted by building two transcriptional units (BBa_K3122004, BBa_ K3122005), and performing Lambda fague and Cell separation assays.

BBa_K3122004 transcriptional unit was assembled in pARK1 alpha plasmid including the following parts:

  • BBa_K3122003: Promoter J23107 adapted to type IIS assembly
  • BBa_K2656009: RBS  B0030 adapted to type IIS assembly
  • BBa_K3122002: coding sequence. This composite part is built with BBa_K3122000 (LamB protein) and BBa_K3122001 (6xHis tag) creating our AEGIS' lamB display system exposing 6xHis Tag.
  • BBa_K2656026: Terminator B0015 adapted to type IIS assembly

BBa_K3122005 transcriptional unit was assembled in pARK1 alpha plasmid including the following parts:

  • BBa_K3122003: Promoter J23107 adapted to type IIS assembly
  • BBa_K2656009: RBS  B0030 adapted to type IIS assembly
  • BBa_K3122000: LamB coding sequence. This part is our AEGIS' lamB display system.
  • BBa_K2656026: Terminator B0015 adapted to type IIS assembly

To see the design of this part go to the Design tab.

How to use it

The only requirement is to adapt the promotor sequence to the MoClo standard, adding the appropiate restriction sites at the ends.

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Parameters
None