Part:BBa_K1401007
Tandem promoter pTet-pBad
This tandem promoter is a combination of pTetR-pBAD and 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 is supposed to be induced by anhydrotetracycline (aTc) and arabinose. 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 (red circles), arabinose alone (blue circles), and then both aTc and arabinose together (purple circles).
As shown below, this tandem promoter functions as expected and is induced with both aTc and arabinose. Fluorescence drops at 5000 and 10000 ng/mL aTc due to a toxicity effect where the E. coli cells died.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 324
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 159
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI site found at 141
| None |