BBa_M36544
1
BBa_M36544
cAMP-activated global transcriptional regulator CRP
2013-10-24T11:00:00Z
2015-05-08T01:14:05Z
http://www.uniprot.org/uniprot/P0ACJ8
cAMP regulatory protein (CRP) binds to cAMP, causes a conformational change, allowing binding to template DNA within the promoter.
false
false
_848_
0
19061
9
Not in stock
false
There was a one codon overlap between the RBS and the protein, so we removed that repeat. We had to find the specific regulatory protein that binds to cAMP.
false
Danielle Fraga
BBa_M36545
1
BBa_M36545
apFAB390 Terminator
2013-10-24T11:00:00Z
2015-05-08T01:14:05Z
BioFab http://io.biofab.org/services/studio/dac/
apFAB390 terminates transcription. It has a 99% Termination Efficiency.
false
false
_848_
0
19061
9
Not in stock
false
We wanted to be sure that transcription was terminated completely.
false
Danielle Fraga
BBa_M36547
1
BBa_M36547
cAMP sensor
2013-12-06T12:00:00Z
2015-05-08T01:14:05Z
The promoter, RBS, and terminator were sourced from Parts Registry.
???BIOFAB Promoter Data: apFAB49.??? BIOFAB, n.d. Web. 24 Oct. 2013. <http://io.biofab.org/services/studio/dac/>.
Mutalik, et. al. ???Precise and reliable gene expression via standard transcription and translation initiation elements.??? Figure 4C. Nature America. 2012. Web. 24 Oct. 2013. <https://coursework.stanford.edu/access/content/group/F13-BIOE-44-01/Required%20readings/Mutalek%202012.pdf
???BIOFAB Terminator Data: apFAB390.??? BIOFAB, n.d. Web. 24 Oct. 2013. <http://io.biofab.org/services/studio/dac/>.
"BIOFAB Promoter Data: apFAB47." BIOFAB, n.d. Web. 24 Oct. 2013. <http://io.biofab.org/services/studio/dac/>.
The CRP coding sequence was sourced from UniProtKB.
???P0ACJ8 (CRP_ECOLI) Reviewed.??? Protein Knowledgebase(UniProtKB). UniProtKB/Swiss-Prot, n.d. Web. 25 Oct. 2013. <http://www.uniprot.org/uniprot/P0ACJ8>.
???Sequence Analysis.??? Reverse Translate. The Sequence Manipulation Suite, n.d. Web. 25 Oct. 2013. <http://www.bioinformatics.org/sms2/rev_trans.html>.
This sensor should detect cAMP in media, activate cAMP regulator protein, and produce a PoPS signal, which then could be the input for an actuator, such as a Gemini reporter.
false
false
_848_
0
19061
9
Not in stock
false
The design behind using both a weak promoter and a weak RBS is intended to keep the E. coli alive since it is known that excessive concentrations of cAMP can be toxic to the cells. However, our second promoter is strong in order to increase our PoPS signal and give high expression of the actuator output.
false
Danielle Fraga, Jason Yang, Kaelo Moahi
component2371601
1
BBa_M36544
component2371602
1
BBa_M36545
component2371599
1
BBa_M36541
component2371600
1
BBa_M36542
component2371603
1
BBa_M36546
annotation2371600
1
BBa_M36542
range2371600
1
48
135
annotation2371603
1
BBa_M36546
range2371603
1
855
904
annotation2371599
1
BBa_M36541
range2371599
1
1
47
annotation2371602
1
BBa_M36545
range2371602
1
766
854
annotation2371601
1
BBa_M36544
range2371601
1
136
765
BBa_M36542
1
BBa_M36542
BCD22 - RBS
2013-10-24T11:00:00Z
2015-05-08T01:14:05Z
Mutalek et. al. 2012
BioFab biofab.org/data (downloaded in csv format)
BCD22 is a weak RBS for translation of RNA into the protein that will express GFP.
false
false
_848_
0
19061
9
Not in stock
false
We had to make sure we did not overexpress cAMP regulatory protein.
false
Danielle Fraga
BBa_M36546
1
BBa_M36546
apFAB47 Promoter with DNA binding sequence from CRP
2013-10-24T11:00:00Z
2015-05-08T01:14:05Z
BioFab http://io.biofab.org/services/studio/dac/
http://www.uniprot.org/uniprot/P0ACJ8
After CRP binds with cAmp, it binds to this specific DNA promoter between the -35 and -10 boxes. It is a strong promoter so that it makes a strong PoPS signal to initiate transcription of an actuator.
false
false
_848_
0
19061
9
Not in stock
false
Between the -35 box and the -10 box, we deleted the sequence from apFAB47 promoter and replaced it with the binding sequence that CRP will bind to.
false
Danielle Fraga
BBa_M36541
1
BBa_M36541
apFAB49 Promoter activated by cAMP in E. Coli
2013-10-24T11:00:00Z
2015-05-08T01:14:05Z
BioFab http://io.biofab.org/services/studio/dac/
apFAB49 is a promoter that gives weak constitutive expression of downstream genes. It initiates transcription, which makes RNA.
The promoter has a 94 Mean Fluorescence per Cell.
false
false
_848_
0
19061
9
Not in stock
false
We wanted a promoter that had weak expression so as to not kill the E. Coli cells with toxicity from overexpression of cAMP receptor protein.
false
Danielle Fraga
BBa_M36547_sequence
1
aaaaagagtattgacttttatcccttgcggcgaatacttacagccatgggcccaagttcacttaaaaaggagatcaacaatgaaagcaattttcgtactgaaacatcttaatcatgcctaggaagttttctaatgatggtgctgggcaaaccgcagaccgatccgaccctggaatggtttctgagccattgccatattcataaatatccgagcaaaagcaccctgattcatcagggcgaaaaagcggaaaccctgtattatattgtgaaaggcagcgtggcggtgctgattaaagatgaagaaggcaaagaaatgattctgagctatctgaaccagggcgattttattggcgaactgggcctgtttgaagaaggccaggaacgcagcgcgtgggtgcgcgcgaaaaccgcgtgcgaagtggcggaaattagctataaaaaatttcgccagctgattcaggtgaacccggatattctgatgcgcctgagcgcgcagatggcgcgccgcctgcaggtgaccagcgaaaaagtgggcaacctggcgtttctggatgtgaccggccgcattgcgcagaccctgctgaacctggcgaaacagccggatgcgatgacccatccggatggcatgcagattaaaattacccgccaggaaattggccagattgtgggctgcagccgcgaaaccgtgggccgcattctgaaaatgctggaagatcagaacctgattagcgcgcatggcaaaaccattgtggtgtatggcacccgctagagatcaagccttaacgaactaagacccccgcaccgaaaggtccgggggttttttttgaccttaaaaacataaccgaggagcagacaaaaaagagtattgactaaatgtgatctagatcacattttaattatttcat
BBa_M36544_sequence
1
atggtgctgggcaaaccgcagaccgatccgaccctggaatggtttctgagccattgccatattcataaatatccgagcaaaagcaccctgattcatcagggcgaaaaagcggaaaccctgtattatattgtgaaaggcagcgtggcggtgctgattaaagatgaagaaggcaaagaaatgattctgagctatctgaaccagggcgattttattggcgaactgggcctgtttgaagaaggccaggaacgcagcgcgtgggtgcgcgcgaaaaccgcgtgcgaagtggcggaaattagctataaaaaatttcgccagctgattcaggtgaacccggatattctgatgcgcctgagcgcgcagatggcgcgccgcctgcaggtgaccagcgaaaaagtgggcaacctggcgtttctggatgtgaccggccgcattgcgcagaccctgctgaacctggcgaaacagccggatgcgatgacccatccggatggcatgcagattaaaattacccgccaggaaattggccagattgtgggctgcagccgcgaaaccgtgggccgcattctgaaaatgctggaagatcagaacctgattagcgcgcatggcaaaaccattgtggtgtatggcacccgc
BBa_M36541_sequence
1
aaaaagagtattgacttttatcccttgcggcgaatacttacagccat
BBa_M36545_sequence
1
tagagatcaagccttaacgaactaagacccccgcaccgaaaggtccgggggttttttttgaccttaaaaacataaccgaggagcagaca
BBa_M36542_sequence
1
gggcccaagttcacttaaaaaggagatcaacaatgaaagcaattttcgtactgaaacatcttaatcatgcctaggaagttttctaatg
BBa_M36546_sequence
1
aaaaagagtattgactaaatgtgatctagatcacattttaattatttcat
igem2sbol
1
iGEM to SBOL conversion
Conversion of the iGEM parts registry to SBOL2.1
James Alastair McLaughlin
Chris J. Myers
2017-03-06T15:00:00.000Z