Composite

Part:BBa_K1608002:Design

Designed by: Ching-Yuan Chen, Yi-Ru Liu, Pei-Hong Chen   Group: iGEM15_Mingdao   (2015-09-10)

GST-SR/pSB1C3


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1338
    Illegal BglII site found at 1407
    Illegal BglII site found at 1524
    Illegal BamHI site found at 798
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 210


Design Notes

  • No EcoRI, XbaI, SpeI, PstI sites on the gene. It is easy to be used as a standard BioBrick part.


We designed two primers to amplify Ptac-GST-SR DNA fragment by PCR from SR/pGEX-2T, followed by XbaI and PstI digestion and ligated to pSB1C3. The plasmid DNA has been checked by colony PCR, restriction enzyme check and sequencing confirmation. The SR gene is fused with GST tag and driven by Ptac promoter along with a lac operator downstream.


Primers:

  • 1.Ptac (pGEX)-XbaI-F: 5'- GAGGGGTCTAGAGTTTTTGCGCCGACATCATAAC -3'
  • 2.SR-SpeI-PstI-R: 5'- AAGAAACTGCAGCGGCCGCTACTAGTATTATGTACGAGAGCGAGA -3'


PCR product size: ~1572 bp


Plasmid map:

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For more DNA gel data & sequencing data, please go to our [http://2015.igem.org/Team:Mingdao/Parts page].

Source

The fragment was cloned from SR/pGEX-2T.

References

A human importin-beta family protein, transportin-SR2, interacts with the phosphorylated RS domain of SR proteins. [http://www.ncbi.nlm.nih.gov/pubmed/10713112 J Biol Chem. 2000]