Composite

Part:BBa_K364330

Designed by: Endre Károly Kristóf   Group: iGEM10_Debrecen-Hungary   (2010-10-20)

TRE-PolyA

Tetracycline Response Element - minimal CMV promoter - PolyA tail which is a fusion of two regulators. It is not possible to fuse coding sequences between them because of scar formation after the ligation but it can be a negative control for the expression vectors made by standard BioBrick parts from pSB1A3 (BBa_K364332, BBa_K364333).

TRE-CMV

The Tetracycline Response Element (TRE) is recognized and bound by the Tetracycline repressor (TetR) protein. The TRE consists of 7 repeats separated by spacer sequences and placed upstream of CMV minimal promoter that has basal expession in the abscence of bound TetR. Tetracycline derivatives (e.g. doxycycline) bind TetR and render it incapable of binding to TRE, thereby forcing the expression of target genes.

PolyA

The PolyA tail of mRNA has multiple adenilates which is important for the nuclear export, translation and stability of mRNA in eukaryotes.

Picture of gel electrophoresis: TRE-CMV and PolyA in pSB1C3 result a 512 bp long insert while the length of TRE-CMV insert is 321 bp.
Picture of gel electrophoresis: TRE-CMV and PolyA in pSB1A3 result a 512 bp long insert forming a plasmid which can be a negative control of the expression vector built up form standard BioBrick parts.

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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