Composite
AraC_TEV-F

Part:BBa_K627008:Design

Designed by: iGEM11 Potsdam_Bioware   Group: iGEM11_Potsdam_Bioware   (2011-09-21)
Revision as of 15:46, 21 September 2011 by UP Sebastian (Talk | contribs) (→‎Source)

Fusion part of arabinose-inducible induction system and the TEV protease


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1144
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1239
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 961
    Illegal SapI.rc site found at 1563


Design Notes

This biobrick was built by PCR using the following PCR primers:

Primer used for site directed mutagenesis:
Fragment 1:

  • r_TEV_ACCAGC: GGAATGTGCAGCTGGTGTCTGACACC
  • f_TEV_iGEM: ATATAGAATTCGCGGCCGCTTCTAGATGGGAGAAAGCTTGTTTAAGGGA

Fragment 2:

  • r_TEV_iGEM_BamHI: ATATAGGATCCACTGCAGCGGCCGCTACTAGTTTATTGCGAGTACACCAATTCATTCAT
  • f_TEV_ACCAGC: GGTGTCAGACACCAGCTGCACATTCC


Primer used for assembly PCR of mutated fragments:

  • f_TEV_iGEM: ATATAGAATTCGCGGCCGCTTCTAGATGGGAGAAAGCTTGTTTAAGGGA
  • r_TEV_iGEM_BamHI: TATAGGATCCACTGCAGCGGCCGCTACTAGTTTATTGCGAGTACACCAATTCATTCAT


Primers used for amplification of pBAD arabinose-inducible induction system:

  • f_AraC_iGEM_HindIII: TATAAGCTTGAATTCGCGGCCGCTTCTAGATTATGACAACTTGACGGCTACATCATT
  • r_AraC_NgoMIV: ATAGCCGGCCTCCTTCTTAAAGTTAAACAAAATTATTTCTAGCCC


Primers used for amplification and modification of TEV protease:

  • f_TEV_AraFusion_NgoMIV: ATATTGCCGGCATGGGAGAAAGCCTGTTTAAGGGA
  • r_TEV_iGEM_BamHI: ATATAGGATCCACTGCAGCGGCCGCTACTAGTTTATTGCGAGTACACCAATTCATTCAT

Source

Arabinose inducible operon form pBAD_iGEM_express, TEV protease from Gunther Stier et. al.

References