Stanford/BIOE44:Module3:Day2:nitrate: Difference between revisions
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==STATUS== | |||
Loaded up at DNA2.0 and authors have confirmed sequence is correct. Good to order! | |||
'''Nitrate Sensor Project''' | '''Nitrate Sensor Project''' | ||
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Novel Bio Bricks Part: Bba_NRL001 | Novel Bio Bricks Part: Bba_NRL001 | ||
'''Long Description:''' | '''Long Description:''' The NarL protein is a cytoplasmic transcription factor that serves as the second component in the two-component Nar pathway found in ''E. Coli'' K-12. When NarX (a membrane protein) binds nitrate, it autophosphorylates and then acts as a histidine kinase on the NarL protein. When NarL is phosphorylated, it binds bacterial DNA and acts as a repressor/activator, depending on the particular operon. | ||
'''Short Description:''' Coding sequence for NarL protein in accordance with BioBrick assembly 10 | '''Short Description:''' Coding sequence for the NarL protein, a response regulator in Nar pathway, in accordance with BioBrick assembly 10. | ||
'''Sequence:''' | '''Sequence:''' | ||
gaattcgcggccgcttctagATGAGTAATCAGGAACCGGCTACTATCCTGCTGATTGACGATCACCCGATGCTGCGAACTGGCGTAAAACAGCTTATCAGTATGGCACCAGATATCACCGTGGTTGGCGAAGCGAGTAATGGCGAACAGGGTATTGAACTGGCGGAGTCTCTTGATCCCGATCTGATCCTGTTAGATCTCAATATGCCCGGCATGAACGGTCTGGAAACGCTGGATAAACTGCGCGAAAAGTCCCTCTCAGGGCGCATTGTGGTATTCAGCGTCTCTAACCATGAAGAAGATGTGGTCACCGCACTGAAACGCGGCGCGGATGGCTATCTGTTAAAAGATATGGAACCGGAAGATCTGCTGAAAGCATTGCATCAGGCAGCTGCTGGCGAAATGGTATTAAGCGAAGCATTAACGCCTGTTCTGGCCGCCAGCTTGCGCGCTAACCGTGCCACTACTGAGCGCGATGTTAACCAGTTAACCCCACGCGAGCGCGATATTCTCAAGCTGATTGCCCAGGGTTTGCCGAACAAGATGATTGCCCGCCGCCTGGATATCACCGAAAGCACAGTAAAAGTGCACGTCAAGCACATGCTGAAGAAAATGAAGCTCAAGTCTCGCGTGGAAGCAGCGGTATGGGTGCATCAGGAGCGCATTTTCTAAtactagtagcggccgctgcag | |||
Natural NarL sequence did not contain any EcoRI, PstI, SpeI, or XbaI restriction sites. NarL part is therefore usable by BioBrick Assmebly 10 standards. | Natural NarL sequence did not contain any EcoRI, PstI, SpeI, or XbaI restriction sites. NarL part is therefore usable by BioBrick Assmebly 10 standards. | ||
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'''Stewart, V., R. S. Rabin''' 1993. Dual Response Regulators (NarL and NarP) Interact with Dual Sensors (NarX and NarQ) To Control Nitrate - and Nitrite - Regulated Gene Expression in ''Escherichia coli'' K-12. J. Bacteriol. '''175:''' 3259 - 3268 | '''Stewart, V., R. S. Rabin''' 1993. Dual Response Regulators (NarL and NarP) Interact with Dual Sensors (NarX and NarQ) To Control Nitrate - and Nitrite - Regulated Gene Expression in ''Escherichia coli'' K-12. J. Bacteriol. '''175:''' 3259 - 3268 | ||
'''Sequence Information''' | |||
Link to [http://www.ncbi.nlm.nih.gov/gene/945795 NarL Sequence Information] | |||
'''Additional Resources''' | |||
after downloading the Ape sequence recognition program, | |||
http://www.biology.utah.edu/jorgensen/wayned/ape/ | |||
we checked the 650 bp sequence for EcoRI, Xba1, SpeI, and Pst1 restriction sites (none) which we send out to be synthesized. | |||
Validated that the part we ordered to be synthesized and the part we designed were the same, | |||
http://workbench.sdsc.edu/ | |||
primers were developed from the following program (which adds biobrick parts), | |||
http://ginkgobioworks.com/cgi/primer.cgi |
Latest revision as of 15:28, 4 May 2010
STATUS
Loaded up at DNA2.0 and authors have confirmed sequence is correct. Good to order!
Nitrate Sensor Project
Authors: Scott Runyon and Travis Urban
Novel Bio Bricks Part: Bba_NRL001
Long Description: The NarL protein is a cytoplasmic transcription factor that serves as the second component in the two-component Nar pathway found in E. Coli K-12. When NarX (a membrane protein) binds nitrate, it autophosphorylates and then acts as a histidine kinase on the NarL protein. When NarL is phosphorylated, it binds bacterial DNA and acts as a repressor/activator, depending on the particular operon.
Short Description: Coding sequence for the NarL protein, a response regulator in Nar pathway, in accordance with BioBrick assembly 10.
Sequence: gaattcgcggccgcttctagATGAGTAATCAGGAACCGGCTACTATCCTGCTGATTGACGATCACCCGATGCTGCGAACTGGCGTAAAACAGCTTATCAGTATGGCACCAGATATCACCGTGGTTGGCGAAGCGAGTAATGGCGAACAGGGTATTGAACTGGCGGAGTCTCTTGATCCCGATCTGATCCTGTTAGATCTCAATATGCCCGGCATGAACGGTCTGGAAACGCTGGATAAACTGCGCGAAAAGTCCCTCTCAGGGCGCATTGTGGTATTCAGCGTCTCTAACCATGAAGAAGATGTGGTCACCGCACTGAAACGCGGCGCGGATGGCTATCTGTTAAAAGATATGGAACCGGAAGATCTGCTGAAAGCATTGCATCAGGCAGCTGCTGGCGAAATGGTATTAAGCGAAGCATTAACGCCTGTTCTGGCCGCCAGCTTGCGCGCTAACCGTGCCACTACTGAGCGCGATGTTAACCAGTTAACCCCACGCGAGCGCGATATTCTCAAGCTGATTGCCCAGGGTTTGCCGAACAAGATGATTGCCCGCCGCCTGGATATCACCGAAAGCACAGTAAAAGTGCACGTCAAGCACATGCTGAAGAAAATGAAGCTCAAGTCTCGCGTGGAAGCAGCGGTATGGGTGCATCAGGAGCGCATTTTCTAAtactagtagcggccgctgcag
Natural NarL sequence did not contain any EcoRI, PstI, SpeI, or XbaI restriction sites. NarL part is therefore usable by BioBrick Assmebly 10 standards.
References:
Stewart, V., H. Lin, P. Bledsoe. 2007. Activation of yeaR-yoaG Operon Transcription by the Nitrate-Responsive Regulator NarL Is Independent of Oxygen-Responsive Regulator Fnr in Escherichia coli K-12. J. Bacteriol. 189: 7539 - 7548
Stewart, V., R. S. Rabin 1993. Dual Response Regulators (NarL and NarP) Interact with Dual Sensors (NarX and NarQ) To Control Nitrate - and Nitrite - Regulated Gene Expression in Escherichia coli K-12. J. Bacteriol. 175: 3259 - 3268
Sequence Information
Link to NarL Sequence Information
Additional Resources
after downloading the Ape sequence recognition program, http://www.biology.utah.edu/jorgensen/wayned/ape/ we checked the 650 bp sequence for EcoRI, Xba1, SpeI, and Pst1 restriction sites (none) which we send out to be synthesized.
Validated that the part we ordered to be synthesized and the part we designed were the same, http://workbench.sdsc.edu/
primers were developed from the following program (which adds biobrick parts), http://ginkgobioworks.com/cgi/primer.cgi