Oligos to Build

Lac System:

Part	Forward	Reverse
###	###	###
###	###	###
###	###	###
###	###	###
###	###	###
###	###	###
###	###	###
###	###	###

Lsr System:

Part	Forward	Reverse
###	###	###
###	###	###
###	###	###
###	###	###
###	###	###
###	###	###
###	###	###
###	###	###

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