Composite

Part:BBa_K3806016:Design

Designed by: Laura Sierra Heras   Group: iGEM21_TUDelft   (2021-09-30)
Revision as of 00:33, 21 October 2021 by LauraSH (Talk | contribs)


Theophylline-binding aptazyme regulating lacZ expression (semi-cRBS). With T7 promoter.


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 3204
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 2321
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

This biobrick represents the DNA template for the expression of the lacZ reporter gene in a T7 based cell-free expression system. Expression is regulated by a theophylline binding aptazyme. The dsRNA sequence was PCR amplified from BBa_K3806014, with a primer that changes the C and T (U in the aptazyme) nucleotides of the antisense strand into G and A, respectively, resulting in a partial mismatch to the RBS (Fig. 1).

Fwd primer: 5’ AATTTAATACGACTCACTATAGGGAAACAAACAAAGACCTTGCTGTCACCGGAATG 3’ Rv primer: 5’ CACACAGGAAACAGCTATGACCATG 3’



References