De novo production of the monoterpenoid geranic acid by metabolically engineered Pseudomonas putida

Table 1 Primers used in this study

Primer	Sequence (5‘ → 3‘)	Source
pJeM1-MCS-f	[phos]AATTCAGGCGCTTTTTAGACTGGTCGTAATGAACCTCTAGAAGTATATTAGTTAAGTATAAGAAGGAGTTTAAACGGTACCGTCGACGAGCTCCTCGAGTGTACAGGATCCA	This work
pJeM1-MCS-r	[phos]AGCTTGGATCCTGTACACTCGAGGAGCTCGTCGACGGTACCGTTTAAACTCCTTCTTATACTTAACTAATATACTTCTAGAGGTTCATTACGACCAGTCTAAAAAGCGCCTG	This work
GES-fwd	GATCGGTACCATGCCTCTAAGTTCAACTC	This work
GES-rev	GATCGAGCTCTTATTGAGTGAAGAAGAGG	This work
pMiS1-hmgs-f	GATCGGATCCAGGAGGAATAATATGAAGAAGCGCGTGGGAATC	This work
pMiS1-hmgs-r	GATCAAGCTT CCTAGGTCAGTTCCCTTCGGCGTAC	This work
pMiS1-MVA-f1	ATCTGGAT CC TAGGAGGAATAATATGGGCGACGACATCACTG	This work
pMiS1-MVA-r1	AACACCATGGCGAGCTCTC	This work
pMiS1-MVA-f2	GAGAGCTCGCCATGGTGTT	This work
pMiS1-MVA-r2	GTGCCCGTTGAGCTCCACCT	This work
pMiS1-MVA-f3	AGGTGGAGCTCAACGGGCAC	This work
pMiS1-MVA-r3	ATCGAATTC AAGCTTTCAGCTCAGCGCGCGCACC	This work

Italics and bold parts represent first and second (if available) restriction site used for cloning, respectively. pJeM1-MCS-f and pJeM1-MCS-r were 5’-phosphorylated ([phos]) and used as sense and antisense DNA strands to exchange egfp of pJeM1 with an insert comprising the rhamnose-inducible promoter rhaP _BAD, a ribosome binding site and an MCS. For more details of the cloning strategy, see Additional file 2.

ISSN: 1475-2859