Difference between revisions of "Part:BBa K2100043"

Latest revision as of 05:21, 19 October 2016

pEXPR pERE5:BM3R1

This construct is an expression vector of a pERE5 promoter upstream of a BM3R1 repressor. pEREx5 is a synthetic mammalian promoter that responds to estrogen receptor activated by estradiol (E2). pEREx5 consists of five repeats of the estrogen response element (ERE) with the sequence 5′-GGTCAnnnTGACC-3 upstream of a minimal promoter (minCMV). We interspaced these elements with 22 randomly selected bases in between, the same spacing between TREs in the TRE promoter. This design choice was made because when an ERE is not properly oriented, it has trouble interacting with ERα and polymerases. Since we wanted to decrease the probability of helical turns inhibiting the transcriptional activity of promoters, we modeled the spacing after the TRE promoter.

In endometrial cells, estrogen receptors facilitate the cell's response to estrogen, initiating the proliferative phase of the menstrual cycle. Mechanistically, estrogen diffuses through the cell membrane and binds to the ligand binding domain of the ER. The DNA-binding domain of the estrogen receptor becomes exposed. The receptor then relocalizes to the nucleus from the cytoplasm, recruits co-activators, and acts as a transcription factor for the BM3R1 gene. When transcribed, BM3R1 represses the DNA sequences found on the pBM3R1 promoter with the help of the corepressor VP16-Gal4.

Sequence and Features

Assembly Compatibility:

10
INCOMPATIBLE WITH RFC[10]
Illegal EcoRI site found at 6
Illegal EcoRI site found at 271
Illegal EcoRI site found at 281
Illegal EcoRI site found at 687
Illegal PstI site found at 504
12
INCOMPATIBLE WITH RFC[12]
Illegal EcoRI site found at 6
Illegal EcoRI site found at 271
Illegal EcoRI site found at 281
Illegal EcoRI site found at 687
Illegal NheI site found at 921
Illegal PstI site found at 504
21
INCOMPATIBLE WITH RFC[21]
Illegal EcoRI site found at 6
Illegal EcoRI site found at 271
Illegal EcoRI site found at 281
Illegal EcoRI site found at 687
23
INCOMPATIBLE WITH RFC[23]
Illegal EcoRI site found at 6
Illegal EcoRI site found at 271
Illegal EcoRI site found at 281
Illegal EcoRI site found at 687
Illegal PstI site found at 504
25
INCOMPATIBLE WITH RFC[25]
Illegal EcoRI site found at 6
Illegal EcoRI site found at 271
Illegal EcoRI site found at 281
Illegal EcoRI site found at 687
Illegal PstI site found at 504
Illegal NgoMIV site found at 426
Illegal NgoMIV site found at 661
Illegal AgeI site found at 439
1000
COMPATIBLE WITH RFC[1000]

@@ Line 3: / Line 3: @@
 <partinfo>BBa_K2100043 short</partinfo>
-description here.
+This construct is an expression vector of a pERE5 promoter upstream of a BM3R1 repressor. pEREx5 is a synthetic mammalian promoter that responds to estrogen receptor activated by estradiol (E2). pEREx5 consists of five repeats of the estrogen response element (ERE) with the sequence 5′-GGTCAnnnTGACC-3 upstream of a minimal promoter (minCMV). We interspaced these elements with 22 randomly selected bases in between, the same spacing between TREs in the TRE promoter. This design choice was made because when an ERE is not properly oriented, it has trouble interacting with ERα and polymerases. Since we wanted to decrease the probability of helical turns inhibiting the transcriptional activity of promoters, we modeled the spacing after the TRE promoter.
+In endometrial cells, estrogen receptors facilitate the cell's response to estrogen, initiating the proliferative phase of the menstrual cycle. Mechanistically, estrogen diffuses through the cell membrane and binds to the ligand binding domain of the ER. The DNA-binding domain of the estrogen receptor becomes exposed. The receptor then relocalizes to the nucleus from the cytoplasm, recruits co-activators, and acts as a transcription factor for the BM3R1 gene. When transcribed, BM3R1 represses the DNA sequences found on the pBM3R1 promoter with the help of the corepressor VP16-Gal4.
 <!-- Add more about the biology of this part here