Regulatory

Part:BBa_K1431503

Designed by: Yongkang Long, Longze Su   Group: iGEM14_SUSTC-Shenzhen   (2014-10-12)

Upstream Activation Sequence(UAS) bind to the GAL4 protein and then activate gene transcription

Upstream Activation Sequence(UAS) is a 17bp sequence: 5'-CGGRNNRCYNYNYNCNCCG-3'(R: Purine, Y: Pyridine N: Any base) that can bind to yeast transcription activator protein GAL4. It usually locates in the upstream of a promoter so that GAL4's activation domain can activate the gene transcription. It usually works as GAL4/UAS system containing two parts: the GAL4 gene and reported gene containing UAS in human cell and Drosophila. Cells contain this system can be detected more green fluorescence than the cells do not if the reporter gene is GFP. We also can use only the DNA binding domain of GAL4. Plasmids contain UAS sequences binding to GAL4. Therefore a chimeric fusion protein containing cell binding domain, translocation domain and DNA binding domain (e.g. GAL4) has the ability to bind the nucleic acid and then transfer that nucleic acid into the cell.


In this situation, the location of UAS should not continue to be in the upstream of a promoter. If the location of UAS does not have influence on the expression of your gene of interest, it can locate at any place but not include within the expression unit.


In our project, we have two chimeric fusion proteins to act as nucleic acid transfer system. One is GD5, containing the human ErbB2 receptor ligand scFv, translocation domain of diphtheria toxin, and GAL4 DNA binding domain. The other is TEG, containing the human EGF receptor ligand TGF-a, translocation domain of Pseudomonas exotoxin A, and GAL4 DNA binding domain．


Plasmids containing UAS are able to bind to GD5 or TEG chimeric fusion protein can be moved into human cells. 2-8 UAS in plasmids are preferable to react with the GD5 or TEG. Therefore, we design three basic parts of 2, 5 and 7 UAS.


Here is part contain 7*UAS. You can adapt this 7*UAS to the plasmids you want to transfer into the cell, if you have chimeric fusion proteins work as nucleic acid transfer system similar to GD5 or TEG.



Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 100
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


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Parameters
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