Device

Part:BBa_K1137011

Designed by: Matthew Deyell   Group: iGEM13_Paris_Bettencourt   (2013-09-23)

sRNA anti lac

Highly similar to BBa_K1137009 and BBa_K1137010, this part contains everything necessary to express small RNA to inhibit expression of lacZ. The biobrick contains a Pr promoter, target sRNA, a scaffold derived from MicC lacking the ompC binding sequence and a T1/TE terminator. The scaffold forms the RNA secondary structure while the target sRNA of the sequence CAGTGAATCCGTAATCATGGT bind to the target sequence of ATGACCATGATTACGGATTCACTG which includes the start codon and the first 24 bp of the ORF within the lacZ mRNA. As in the previously described biobricks, it is vital that the sequences are complementary for proper repression to occur. If there are point mutations within the lacZ gene or biobrick sequence the biobrick will not function correctly. Additionally, there will be some leakage of β-Galactosidase which will cause cleavage of X-Gal, however we observed that edges of colonies tended to be white while the center of colonies tended to be blue.

Anti-Lac_PR.png

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 153
    Illegal XhoI site found at 159
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
None