Difference between revisions of "Oligos to Build"
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!width="50"|Oligo Design | !width="50"|Oligo Design | ||
|- | |- | ||
| − | |lpLas'|| | + | |lpLas'|| [[Image:pLasOligos.jpg]] || [[Image:pLas' oligos diagram.jpg]] |
|- | |- | ||
| − | |pLasXOR|| | + | |pLasXOR|| [[Image:pLasXOR Oligos.jpg]] || [[Image:pLasXOR.jpg]] |
|} | |} | ||
Latest revision as of 20:32, 20 June 2008
Lac System:
| Part | Forward | Reverse |
|---|---|---|
| luxS |  |
|
| lacIQ1 Promoter |  |
|
| lacIQ Promoter |  |
|
| LacI_I12 |  |
|
| LacI |  |
|
| LacI_X86 (Primers for step 1 of PCR) | |
|
| LacI_I12X86 (Primers for step 1 of PCR) | |
|
| ### | ### | ### |
Lsr System:
| Part | Forward | Reverse |
|---|---|---|
| lsrR |  |
|
| lsrK |  |
|
| plsr |  |
|
| ### | ### | ### |
| ### | ### | ### |
| ### | ### | ### |
| ### | ### | ### |
| ### | ### | ### |
Davidson XOR Promoters:
| Part | Oligos | Oligo Design |
|---|---|---|
| lpLas' |  |
|
| pLasXOR |  |
|
Sequences of Parts to Build
LuxS Wildtype from MG1655
ATGCCGTTGT TAGATAGCTT CACAGTCGAT CATACCCGGA TGGAAGCGCC TGCAGTTCGG GTGGCGAAAA CAATGAACAC CCCGCATGGC GACGCAATCA CCGTGTTCGA TCTGCGCTTC TGCGTGCCGA ACAAAGAAGT GATGCCAGAA AGAGGGATCC ATACCCTGGA GCACCTGTTT GCTGGTTTTA TGCGTAACCA TCTTAACGGT AATGGTGTAG AGATTATCGA TATCTCGCCA ATGGGCTGCC GCACCGGTTT TTATATGAGT CTGATTGGTA CGCCAGATGA GCAGCGTGTT GCTGATGCCT GGAAAGCGGC AATGGAAGAC GTGCTGAAAG TGCAGGATCA GAATCAGATC CCGGAACTGA ACGTCTACCA GTGTGGCACT TACCAGATGC ACTCGTTGCA GGAAGCGCAG GATATTGCGC GTAGCATTCT GGAACGTGAC GTACGCATCA ACAGCAACGA AGAACTGGCA CTGCCGAAAG AGAAGTTGCA GGAACTGCAC ATCTAG
Map Position: 2,812,240 <- 2,812,755 (nucleotides)
Sequence Length: 516 bp / 171 AAs
Mutated LuxS
Nucleotide position 54 changed from A to T in order to destroy a PstI site; this is a sense mutation
Also changed stop codon from TAG to TAA (recommended in Registry Help)
ATGCCGTTGT TAGATAGCTT CACAGTCGAT CATACCCGGA TGGAAGCGCC TGCTGTTCGG
GTGGCGAAAA CAATGAACAC CCCGCATGGC GACGCAATCA CCGTGTTCGA TCTGCGCTTC
TGCGTGCCGA ACAAAGAAGT GATGCCAGAA AGAGGGATCC ATACCCTGGA GCACCTGTTT
GCTGGTTTTA TGCGTAACCA TCTTAACGGT AATGGTGTAG AGATTATCGA TATCTCGCCA
ATGGGCTGCC GCACCGGTTT TTATATGAGT CTGATTGGTA CGCCAGATGA GCAGCGTGTT
GCTGATGCCT GGAAAGCGGC AATGGAAGAC GTGCTGAAAG TGCAGGATCA GAATCAGATC
CCGGAACTGA ACGTCTACCA GTGTGGCACT TACCAGATGC ACTCGTTGCA GGAAGCGCAG
GATATTGCGC GTAGCATTCT GGAACGTGAC GTACGCATCA ACAGCAACGA AGAACTGGCA
CTGCCGAAAG AGAAGTTGCA GGAACTGCAC ATCTAA
LsrR Gene
ATGACAATCA ACGATTCGGC AATTTCAGAA CAGGGAATGT GTGAAGAAGA ACAGGTCGCG CGGATCGCGT GGTTTTACTA TCACGACGGG CTGACCCAGA GCGAGATCAG CGATCGTCTC GGCCTGACAC GTTTGAAAGT GTCGCGATTG CTGGAGAAAG GGCATCAGTC CGGCATTATT CGCGTACAGA TTAATTCTCG CTTTGAAGGC TGTCTGGAAT ATGAAACTCA ATTACGTCGT CAGTTTTCGC TGCAACATGT CCGGGTGATC CCTGGGCTTG CGGATGCTGA TGTCGGTGGG CGACTGGGGA TAGGCGCGGC GCATATGTTG ATGAGTTTAC TTCAACCACA ACAGATGCTG GCGATTGGTT TTGGCGAGGC AACCATGAAT ACGCTGCAAC GCTTAAGTGG TTTTATTTCG TCACAGCAAA TTCGCCTGGT CACGCTCTCC GGTGGCGTCG GTTCTTATAT GACGGGAATC GGGCAGCTTA ACGCGGCGTG CAGTGTGAAT ATTATTCCGG CTCCGTTGCG GGCATCCTCC GCTGACATTG CCCGTACGCT AAAAAATGAA AATTGCGTCA AAGATGTTCT GTTAGCCGCG CAAGCAGCGG ATGTGGCGAT TGTCGGCATT GGTGCTGTGA GTCAACAGGA CGATGCGACA ATCATTCGCT CCGGTTATAT CAGCCAGGGC GAACAGTTAA TGATTGGCCG AAAAGGGGCG GTTGGCGACA TTTTAGGCTA CTTTTTTGAT GCAAAAGGTG ACGTTGTCAC GAATATCAAA ATACATAACG AACTGATTGG CTTACCTTTA AGCGCGCTGA AGACCATACC CGTCCGGGTT GGCGTGGCAG GGGGAGAAAA TAAAGCCGAA GCAATTGCCG CTGCAATGAA AGGCGGTTAT ATCAACGCAC TGGTTACCGA TCAGGACACA GCAGCGGCGA TTTTACGTAG TTAA
Map Position: 1,598,312 <- 1,599,265 (nucleotides)
Sequence Length: 954bp/317 AAs
LsrK Gene
atgGCTCGAC TCTTTACCCT TTCAGAATCA AAGTACTACC TGATGGCGCT GGATGCAGGC ACCGGAAGTA TTCGGGCTGT GATATTCGAC CTGGAAGGCA ATCAAATAGC AGTGGGACAG GCGGAGTGGC GGCATCTGGC AGTACCGGAC GTTCCTGGTT CTATGGAATT TGATCTCAAC AAAAACTGGC AACTGGCGTG TGAGTGTATG CGCCAGGCGC TGCACAACGC CGGCATAGCC CCGGAGTATA TCGCTGCCGT TTCGGCATGT TCGATGCGTG AAGGCATTGT TTTATATAAT AATGAAGGAG CCCCGATCTG GGCCTGCGCC AATGTGGATG CCAGAGCGGC ACGCGAAGTT AGCGAACTTA AAGAACTGCA CAACAATACC TTTGAAAACG AAGTTTATCG CGCGACCGGA CAAACACTGG CTTTAAGTGC CATCCCCAGA TTACTTTGGC TGGCGCACCA TCGTTCCGAT ATTTACCGTC AGGCATCAAC CATCACCATG ATCAGCGACT GGCTGGCCTA TATGCTCAGC GGCGAACTGG CGGTGGATCC CTCTAACGCT GGCACCACGG GACTTCTTGA TCTAACCACC CGTGACTGGA AACCTGCATT GCTGGATATG GCTGGCCTAC GTGCCGATAT TCTTTCTCCT GTCAAAGAAA CCGGCACATT GCTGGGCGTG GTAAGTTCAC AAGCGGCGGA ACTCTGCGGT CTGAAGGCGG GCACTCCGGT GGTCGTTGGA GGAGGCGACG TGCAGCTTGG TTGCCTTGGG TTAGGCGTTG TGCGTCCGGC ACAAACCGCG GTTCTTGGCG GCACATTCTG GCAGCAAGTT GTAAATTTAG CCGCGCCGGT GACAGACCCA GAAATGAACG TGCGCGTTAA TCCTCATGTT ATTCCTGGCA TGGTACAAGC TGAATCTATA AGCTTTTTTA CCGGACTCAC CATGCGCTGG TTCCGCGATG CTTTCTGTGC CGAAGAAAAA CTGATTGCCG AACGTTTAGG CATCGACACC TATACGCTGC TGGAAGAGAT GGCCAGTCGG GTGCCGCCTG GGTCGTGGGG CGTAATGCCG ATCTTCTCCG ACAGAATGCG CTTTAAAACC TGGTATCACG CTGCGCCTTC CTTTATTAAC TTGTCCATTG ACCCGGATAA ATGTAACAAA GCGACATTGT TCCGTGCGCT GGAAGAAAAT GCGGCGATTG TATCAGCGTG TAACTTGCAG CAAATTGCTG ATTTCTCGAA TATTCATCCT TCATCGTTAG TCTTTGCAGG CGGAGGTTCA AAAGGGAAAT TATGGAGTCA AATTCTCGCT GATGTCTCGG GATTACCCGT CAATATTCCG GTGGTCAAAG AAGCCACTGC ATTAGGATGT GCCATTGCAG CTGGCGTCGG TGCCGGAATT TTTTCATCAA TGGCAGAAAC CGGAGAACGC CTGGTTCGCT GGGAACGGAC GCACACACCA GACCCGGAAA AGCATGAACT TTATCAGGAT TCACGCGATA AGTGGCAGGC AGTTTATCAG GATCAGCTGG GGCTGGTTGA TCATGGACTG ACGACGTCGT TATGGAAAGC GCCTGGGTTA TAG
Map Position: 1,596,641 <- 1,598,233 (nucleotides)
Sequence Length: 1593 bp / 530 AAs
lsr promoter sequence
LacI wild-type gene sequence (ORF begins at the first amino acid)
ATGAAACCAGTAACGTTATACGATGTCGCAGAGTATGCCGGTGTCTCTTATCAGACCGTTTCCCGCGTGGTGAACCAGGCCAGCCACGTTTCTGCGAAAACGCGGGAAAAAGTGGAAGC
GGCGATGGCGGAGCTGAATTACATTCCCAACCGCGTGGCACAACAACTGGCGGGCAAACAGTCGTTGCTGATTGGCGTTGCCACCTCCAGTCTGGCCCTGCACGCGCCGTCGCAA
ATTGTCGCGGCGATTAAATCTCGCGCCGATCAACTGGGTGCCAGCGTGGTGGTGTCGATGGTAGAACGAAGCGGCGTCGAAGCCTGTAAAGCGGCGGTGCACAATCTTCTCGCGC
AACGCGTCAGTGGGCTGATCATTAACTATCCGCTGGATGACCAGGATGCCATTGCTGTGGAAGCTGCCTGCACTAATGTTCCGGCGTTATTTCTTGATGTCTCTGACCAGACACCATCA
ACAGTATTATTTTCTCCCATGAAGACGGTACGCGACTGGGCGTGGAGCATCTGGTCGCATTGGGTCACCAGCAAATCGCGCTGTTAGCGGGCCCATTAAGTTCTGTCTCGGCG
CGTCTGCGTCTGGCTGGCTGGCATAAATATCTCACTCGCAATCAAATTCAGCCGATAGCGGAACGGGAAGGCGACTGGAGTGCCATGTCCGGTTTTCAACAAACCATGCAAATGC
TGAATGAGGGCATCGTTCCCACTGCGATGCTGGTTGCCAACGATCAGATGGCGCTGGGCGCAATGCGCGCCATTACCGAGTCCGGGCTGCGCGTTGGTGCGGATATCTCGGTAGT
GGGATACGACGATACCGAAGACAGCTCATGTTATATCCCGCCGTTAACCACCATCAAACAGGATTTTCGCCTGCTGGGGCAAACCAGCGTGGACCGCTTGCTGCAACTCTCTCAG
GGCCAGGCGGTGAAGGGCAATCAGCTGTTGCCCGTCTCACTGGTGAAAAGAAAAACCACCCTGGCGCCCAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGC
AGCTGGCACGACAGGTTTCCCGACTGGAAAGCGGGCAGTGA
LacI_I12 Mutation (amino acid 3 is changed from CCA to TAT)
ATGAAATATGTAACGTTATACGATGTCGCAGAGTATGCCGGTGTCTCTTATCAGACCGTTTCCCGCGTGGTGAACCAGGCCAGCCACGTTTCTGCGAAAACGCGGGAAAAAGTGGAAGC
GGCGATGGCGGAGCTGAATTACATTCCCAACCGCGTGGCACAACAACTGGCGGGCAAACAGTCGTTGCTGATTGGCGTTGCCACCTCCAGTCTGGCCCTGCACGCGCCGTCGCAA
ATTGTCGCGGCGATTAAATCTCGCGCCGATCAACTGGGTGCCAGCGTGGTGGTGTCGATGGTAGAACGAAGCGGCGTCGAAGCCTGTAAAGCGGCGGTGCACAATCTTCTCGCGC
AACGCGTCAGTGGGCTGATCATTAACTATCCGCTGGATGACCAGGATGCCATTGCTGTGGAAGCTGCCTGCACTAATGTTCCGGCGTTATTTCTTGATGTCTCTGACCAGACACCC
ATCAACAGTATTATTTTCTCCCATGAAGACGGTACGCGACTGGGCGTGGAGCATCTGGTCGCATTGGGTCACCAGCAAATCGCGCTGTTAGCGGGCCCATTAAGTTCTGTCTCGGC
GCGTCTGCGTCTGGCTGGCTGGCATAAATATCTCACTCGCAATCAAATTCAGCCGATAGCGGAACGGGAAGGCGACTGGAGTGCCATGTCCGGTTTTCAACAAACCATGCAAATG
CTGAATGAGGGCATCGTTCCCACTGCGATGCTGGTTGCCAACGATCAGATGGCGCTGGGCGCAATGCGCGCCATTACCGAGTCCGGGCTGCGCGTTGGTGCGGATATCTCGGTAG
TGGGATACGACGATACCGAAGACAGCTCATGTTATATCCCGCCGTTAACCACCATCAAACAGGATTTTCGCCTGCTGGGGCAAACCAGCGTGGACCGCTTGCTGCAACTCTCTCA
GGGCCAGGCGGTGAAGGGCAATCAGCTGTTGCCCGTCTCACTGGTGAAAAGAAAAACCACCCTGGCGCCCAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATG
CAGCTGGCACGACAGGTTTCCCGACTGGAAAGCGGGCAGTGA
LacI_X86 Mutation (amino acid 61 is changed from TCG to CTG)
ATGAAACCAGTAACGTTATACGATGTCGCAGAGTATGCCGGTGTCTCTTATCAGACCGTTTCCCGCGTGGTGAACCAGGCCAGCCACGTTTCTGCGAAAACGCGGGAAAAAGTGGAAGC
GGCGATGGCGGAGCTGAATTACATTCCCAACCGCGTGGCACAACAACTGGCGGGCAAACAGCTGTTGCTGATTGGCGTTGCCACCTCCAGTCTGGCCCTGCACGCGCCGTCGCAA
ATTGTCGCGGCGATTAAATCTCGCGCCGATCAACTGGGTGCCAGCGTGGTGGTGTCGATGGTAGAACGAAGCGGCGTCGAAGCCTGTAAAGCGGCGGTGCACAATCTTCTCGCGC
AACGCGTCAGTGGGCTGATCATTAACTATCCGCTGGATGACCAGGATGCCATTGCTGTGGAAGCTGCCTGCACTAATGTTCCGGCGTTATTTCTTGATGTCTCTGACCAGACACCC
ATCAACAGTATTATTTTCTCCCATGAAGACGGTACGCGACTGGGCGTGGAGCATCTGGTCGCATTGGGTCACCAGCAAATCGCGCTGTTAGCGGGCCCATTAAGTTCTGTCTCGGC
GCGTCTGCGTCTGGCTGGCTGGCATAAATATCTCACTCGCAATCAAATTCAGCCGATAGCGGAACGGGAAGGCGACTGGAGTGCCATGTCCGGTTTTCAACAAACCATGCAAATG
CTGAATGAGGGCATCGTTCCCACTGCGATGCTGGTTGCCAACGATCAGATGGCGCTGGGCGCAATGCGCGCCATTACCGAGTCCGGGCTGCGCGTTGGTGCGGATATCTCGGTAG
TGGGATACGACGATACCGAAGACAGCTCATGTTATATCCCGCCGTTAACCACCATCAAACAGGATTTTCGCCTGCTGGGGCAAACCAGCGTGGACCGCTTGCTGCAACTCTCTCA
GGGCCAGGCGGTGAAGGGCAATCAGCTGTTGCCCGTCTCACTGGTGAAAAGAAAAACCACCCTGGCGCCCAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATG
CAGCTGGCACGACAGGTTTCCCGACTGGAAAGCGGGCAGTGA
LacI_I12_X86 Mutation
ATGAAATATGTAACGTTATACGATGTCGCAGAGTATGCCGGTGTCTCTTATCAGACCGTTTCCCGCGTGGTGAACCAGGCCAGCCACGTTTCTGCGAAAACGCGGGAAAAAGTGGAAGCGGCGATGGCGGAG
CTGAATTACATTCCCAACCGCGTGGCACAACAACTGGCGGGCAAACAGCTGTTGCTGATTGGCGTTGCCACCTCCAGTCTGGCCCTGCACGCGCCGTCGCAAATTGTCGCGGCGATTAAATCTCGCGCC
GATCAACTGGGTGCCAGCGTGGTGGTGTCGATGGTAGAACGAAGCGGCGTCGAAGCCTGTAAAGCGGCGGTGCACAATCTTCTCGCGCAACGCGTCAGTGGGCTGATCATTAACTATCCGCTGGATGAC
CAGGATGCCATTGCTGTGGAAGCTGCCTGCACTAATGTTCCGGCGTTATTTCTTGATGTCTCTGACCAGACACCCATCAACAGTATTATTTTCTCCCATGAAGACGGTACGCGACTGGGCGTGGAGCATC
TGGTCGCATTGGGTCACCAGCAAATCGCGCTGTTAGCGGGCCCATTAAGTTCTGTCTCGGCGCGTCTGCGTCTGGCTGGCTGGCATAAATATCTCACTCGCAATCAAATTCAGCCGATAGCGGAACGGG
AAGGCGACTGGAGTGCCATGTCCGGTTTTCAACAAACCATGCAAATGCTGAATGAGGGCATCGTTCCCACTGCGATGCTGGTTGCCAACGATCAGATGGCGCTGGGCGCAATGCGCGCCATTACCGAGT
CCGGGCTGCGCGTTGGTGCGGATATCTCGGTAGTGGGATACGACGATACCGAAGACAGCTCATGTTATATCCCGCCGTTAACCACCATCAAACAGGATTTTCGCCTGCTGGGGCAAACCAGCGTGGACC
GCTTGCTGCAACTCTCTCAGGGCCAGGCGGTGAAGGGCAATCAGCTGTTGCCCGTCTCACTGGTGAAAAGAAAAACCACCCTGGCGCCCAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCAT
TAATGCAGCTGGCACGACAGGTTTCCCGACTGGAAAGCGGGCAGTGA
LacI Promoter
CGTTGACACCATCGAATGGCGCAAAACCTTTCGCGGTATGGCATGATAGCGCCCGG
LacIQ Promoter
CGTTGACACCATCGAATGGTGCAAAACCTTTCGCGGTATGGCATGATAGCGCCCGG
LacIQ1 Promoter
AGCGGCATGCATTTACGTTGACACCACCTTTCGCGGTATGGCATGATAGCGCCCGG
-These promoter sequences are taken from Glascock and Weickert 1998
TrpR Operator Sequence Only
ATATGCTATCGTACTCTTTAGCGAGTACAACCGGGGGA
