Translational_Unit

Part:BBa_K1223005:Design

Designed by: Orr Schlesinger   Group: iGEM13_BGU_Israel   (2013-09-07)
Revision as of 12:59, 12 October 2013 by Alex m (Talk | contribs) (→‎Design notes)

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cI translational unit


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
    COMPATIBLE WITH RFC[1000]

Design notes

This part was designed to function as a biological timer for our P.A.S.E 1 system. It includes an assembly of two existing parts from the Registry, Lac/Ara-1 IPTG Inducible promoter (BBa_K354000) and LVA-tagged cI repressor protein (BBa_K327018). In order to extend the half life of the cI protein, we removed the LVA tail. The part was characterized through sequencing and restriction digest with PvuII and HindIII.


CI1.png

lane1: undigested pUC57-cI

lane2: pUC57-cI digested with PvuII and HindIII

lane3: DS5000 ladder


In addition, the promoter's performance under the presence of IPTG and Arabinose was analyzed via comassie staining. The cI protein is 29 KDa. in order to characterize the behaviour of this promoter under different induction conditions we built a construct comprised of this promoter, strong RBS and the cI CDS. the promoter's performance under the presence of IPTG and Arabinose was analyzed via comassie staining. The cI protein is 29 KDa. the cI expession levels were dependent on the combination of inducers used to express the cI. this way we generated a 4-way switch to control the amount of protein the cell produces.

CI2.jpg

Source

genebank, iGEM parts registry

References