Part:BBa_K779200
Hef1A MammoBlock
Promoter Vector containing human elongation factor 1a (hEF1a) promoter; acting as a low level constitutive promoter.
We have tested this promoter with many different fluorescent proteins (TagBFP, eYFP, mKate) as well as activators. We have used the Gateway method to create composite parts with the Hef1A promoter: Hef1A:eYFP - BBa_K779600, Hef1A:mKate - BBa_K779601, Hef1A:TagBFP-4xFF5 - BBa_K779602, Hef1A:eYFP-4xFF1 - BBa_K779603, Hef1A:rtTA3 - BBa_K779604
Testing the Hef1A promoter with eYFP and with mKate:
This figure shows HEK293 cells which were transiently transfected with Hef1A:TagBFP and Hef1A:mKate using Lipofectamine 2000. Equimole amounts of DNA were delivered, 500 ng total per 1.65 uL of reagent. Images were taken on a Zeiss microscope at 10X. (a) brightfield (b) overlay of blue, red and brightfield (c) blue channel (d) red channel
Testing the Hef1A promoter with rtTA3 and with TagBFP:
100,000 HEK293 cells were transfected with equimolar rations of Hef1a:rTTA3, TreT:mKate, and Hef1a:TagBFP (as a transfection marker), standardized to 500 ng total plasmid DNA, with 1.65 uL Lipofectamine 2000. After 48 hours, cells were imaged on a Leica Confocal Microscope at 10x. We can see that we have transfection, since cells are fluorescing blue. Also, we can see that as we increase the concentration of DOX present, we see an increase in red fluorescence.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 315
Illegal PstI site found at 820 - 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 315
Illegal PstI site found at 820 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 569
Illegal XhoI site found at 968 - 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 315
Illegal PstI site found at 820 - 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 315
Illegal PstI site found at 820
Illegal NgoMIV site found at 703
Illegal AgeI site found at 81 - 1000COMPATIBLE WITH RFC[1000]
| n/a | Hef1A MammoBlock |