Regulatory

Part:BBa_K1051303

Designed by: Kequan Lin   Group: iGEM13_Shenzhen_BGIC_ATCG   (2013-06-30)

clb6 promoter (during G1 phase)

Purpose

Works to activates Cdc28p to promote initiation of DNA synthesis; functions in formation of mitotic spindles along with Clb3p and Clb4p.

Principle

"clb6" is a promoter which promotes transciption in G1/S of the yeast cell cycle .If you want to express your proteins in G1/S of the yeast cell cycle ,you can add the sequence which is actived by clb6 before your gene sequence ,then put them in a yeast plasmid .After transformation ,you can get the protein you want from the metabolic product of the Bacterial you use for transformation .it most abundant during late G1; CLB6 has a paralog, CLB5, that arose from the whole genome duplication

Measurement

Using the GFP as reporter and morphological alteration as cell cycle representation, we verified the Clb6 can be activated in G1 phase in the yeast. As shown in the picture in normal lights, there are some yeast were budding. These budded yeast cells and small size budding yeasts (within the red circles) are assumed in the G1 phase of cell cycle. And the next picture was taken under activation light, thus the green lights can representing the Clb6 promoter expressing. The two pictures indicated the Clb6 were expressed in the G1 phase as we expected when designing the experiments PS: we modified the contrast ratio to lower the lights of neighbor cells, thus our results looking better. Clb6.jpg Figure. clb6 promoter for G1 phase has been verified.


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]


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Categories
Parameters
n/aclb6 promoter (during G1 phase)