RNA

Part:BBa_K1465225:Design

Designed by: iGEM-Team Bielefeld 2014   Group: iGEM14_Bielefeld-CeBiTec   (2014-10-05)


Knock-down of phosphofructokinase A using sRNA (sRNA:pfkA)


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 104
    Illegal XhoI site found at 110
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

  • Amplification of the pSB1C3-Backbone (BBa_J04450) using the specific Gibson-Primer [http://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Primer#pSB1C3_pre_sRNApfkA pSB1C3_pre_sRNApfkA] and [http://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Primer#pSB1C3_suf_sRNApfkA pSB1C3_suf_sRNApfkA]:
pSB1C3_pre_sRNApfkA (43 bp)

5'-TGTGGCTAAAAGAATTAGTACTCTAGAAGCGGCCGCGAATTCC-3'

pSB1C3_suf_sRNApfkA (43 bp)

5'-TGGGCCTTTCTGCGTTTATATACTAGTAGCGGCCGCTGCAGTC-3'
  • [http://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Protocols#Gibson Gibson-Assembly] of the pSB1C3-Backbone and the gBlock containing the antisense RNA of the Phosphofructoskinase (pfkA) of Escherichia coli.

Source

Gensynthesis ([http://eu.idtdna.com/pages/products/genes/gblocks-gene-fragments gBlocks]) of the sRNA with the anti-sense RNA for the phosphofructokinase (pfkA) of Escherchia coli.

References

  • Dokyun Na, Seung Min Yoo, Hannah Chung, Hyegwon Park, Jin Hwan Park and Sang Yup Lee (2013) Metabolic engineering of

Escherichia coli using synthetic small regulatory RNAs [http://www.nature.com/nbt/journal/v31/n2/pdf/nbt.2461.pdf Nature Biotechnology 31:170 - 174.]