Difference between revisions of "Part:BBa K1223005"

Latest revision as of 12:50, 12 October 2013

cI translational unit

Lambda cI translational unit with BBa_K354000 as promoter. this translational unit is on the other hand very tightly regulated due to the AraC/LacI promoter and on the other hand, produces a large amount of protein when induced by one or both of the inducers.

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.

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. 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.

Source

genebank, iGEM parts registry

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
COMPATIBLE WITH RFC[1000]

@@ Line 2: / Line 2: @@
 <partinfo>BBa_K1223005 short</partinfo>
-Lambda cI translational unit with BBa_K354000 as promoter
+Lambda cI translational unit with BBa_K354000 as promoter. this translational unit is on the other hand very tightly regulated due to the  AraC/LacI promoter and on the other hand, produces a large amount of protein when induced by one or both of the inducers.
@@ Line 21: / Line 21: @@
-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 addition, 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.
 [[File:cI2.jpg]]