Team:UTK-Knoxville/primers

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Team:UK-Knoxville - 2013.igem.org

Gibson Primers

Name Sequence Melting Temperature Function / Comment
Transmembrane Construct
EnvZ_F gtaagtcacgacttgcgcacgccgctgac 70 Amplifies EnvZ does not include any overlap of the sensor part.
Envz_R ggcggccgctctattagtgatggtggtgatgatgttacccttcttttgtcgtgc 75 Amplifies EnvZ (20 bp) and includes overlap (34 bp) of the free space between it and the terminator.
OmpR_F agaaataattttgtttaactataagaaggagatatacatatgcaagagaactacaagatt 64 Amplifies OmpR and overlaps with the RBS and T7 promoter.  Note the backbone needs to have the T7 promoter and RBS already.
OmpR_R tcatgctttagagccgtccggtacaaagac 65 Amplifies OmpR and does not include the second RBS to avoid homology.
 
Aer  
Aer_f tctttgtaccggacggctctaaagcatgaaaggagatataatgtcttctcatccgtatgt 71 Includes overlap with OmpR (29 bp), the RBS (11 bp) and overlap with Aer (20 bp)
Aer_R TACGCgtcagcggcgtgcgcaagtcgtgacttacatttctgacactggacacctg 77 Include overlap with EnvZ (34 bp) and Aer (21 bp).  The temperature differance is quite extereme.
NarX  
NarX_F tctttgtaccggacggctctaaagcatgaAAGGAGATATAatgcttaaacgttgtctctc 71 Includes overlap with OmpR (29 bp), the RBS (11 bp) and overlap with NarX (20 bp)
NarX_R tacgcgtcagcggcgtgcgcaagtcgtgacttacatttttatgctccagcccggc 79 Include overlap with EnvZ (34 bp) and NarX (21 bp).  The temperature differance is quite extereme.
Cytoplasmic Construct
NtrC_F TAGAAATAATTTTGTTTAACTataagaaggagatatacatatgcaacgagggatagtctg 65 Amplifies NtrC and overlaps with the RBS and T7 promoter.  Note the backbone needs to have the T7 promoter and RBS already.
NtrC_R gagctggggatggagtgaTTTTAACTTACGC 64 Amplifies NtrC and does not include the second RBS to avoid homology.
NtrB_F ccgatggataaccagcgccgcTTAAGTCAGG 69 Amplifies NtrB does not include any overlap of the sensor part.
NtrB_R GGCGGCCGCTCTATTAGTGATGGTGGTGATGATGttatttcctgataggcaggt 74 Amplifies NtrB (20 bp) and includes overlap (34 bp) of the free space between it and the terminator.
HemAT
HemAT_F gtaagttaaaatcactccatccccagctcAAGGAGATATAttgttatttaaaaaagacag 67 Includes overlap with NtrC (29 bp), the RBS (11 bp) and overlap with HemAT (20 bp)
HemAT_R CTGTTCCTGACTTAAGCggcgctggttatccatcggctggttgtactcgctttgaaac 75 Include overlap with NtrB (34 bp) and HemAT (21 bp).  The temperature differance is quite extereme.
RcoM
RcoM_F gtaagttaaaatcactccatccccagctcAAGGAGATATAatgtttaaatattttgcaatt 67 Includes overlap with NtrC (29 bp), the RBS (11 bp) and overlap with RcoM (20 bp)
RcoM_R gttcctgacttaagcggcgctggttatccatcggcttatggacggttacactacc 75 Include overlap with NtrB (34 bp) and RcoM (21 bp).  The temperature differance is quite extereme.
TodS
TodS_F gtaagttaaaatcactccatccccagctcAAGGAGATATAatgagctccttggatagaaa 70 Includes overlap with NtrC (29 bp), the RBS (11 bp) and overlap with TodS (20 bp)
TodS_R GTTCCTGACTTAAgcggcgctggttatccatcggatcggtaatattgcgcccttc 75 Include overlap with NtrB (34 bp) and TodS (21 bp)
LasR      
LasR_F      
LasR_R      
      Note: APE and IDT primer Tm differ

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