Regulatory

Part:BBa_K1401007

Designed by: Kathleen Lewis   Group: iGEM14_BostonU   (2014-10-09)
Revision as of 00:17, 14 October 2014 by THaddock (Talk | contribs)

Tandem promoter pTet-pBad

This tandem promoter was created using the MoClo assembly method. This is a Level 0 MoClo part with flanking sites A on the 5' side and site B on the 3' side of the part. The fusion site letters refer to 4bp fusion sites: A = GGAG; B = TACT; C = AATG; D = AGGT; E = GCTT; F = CGCT; G = TGCC; H = ACTA. This tandem promoter is in pSB1C3.

The promoter can be induced by anhydrotetracycline (aTc) and arabinose, as shown below. In order to test this tandem promoter, we built a basic transcriptional unit with red fluorescent protein reporter. We used the E. coli DH5-alpha Pro strain for this study, which has tetR, lacI, and araC all constitutively expressed in its genome. We induced the E. coli with aTc alone, arabinose alone, and then both aTc and arabinose together to see if we could determine if one promoter was dominant over the other.

K1401007 flow.png

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 324
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 159
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 141


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Categories
Parameters
None