Table 1 Primers and plasmids used in this study
From: Engineering of Escherichia coli for Krebs cycle-dependent production of malic acid
Primers | Relevant characteristics | Source |
|---|---|---|
ppc_clon_for | TATAATCCCGGGATGAACGAACAATATTCC | This study |
ppc_clon_for | TATAATTCTAGATTAGCCGGTATTACGCAT | This study |
ppc_k620s_for | CGCTTTAGCTATGGTCTGCCAGAAATCACCGAG | This study |
ppc_k620s_rev | CCATAGCTAAAGCGGATCATCTCGCCC | This study |
ppc_sRBS_for | AACAGAATTCGAGCTCGGTACCCGGGGTTTAACTTTAAGAAGGAGATATACCATGAACGAACAATATTCCGCATTGCGTAGTAATG | This study |
ppc_sRBS_rev | TATAATTCTAGAATTAGCCGGTATTACGCATACC | This study |
gltA_clon_for | TATATAGAGCTCATGGCTGATACAAAAGCAAAACTCACC | This study |
gltA_clon_rev | TATAATAAGCTTTTAACGCTTGATATCGCTTTTAAAGTCGC | This study |
gltA_R164L_for | ATTGCCGCGTTCCTCCTGCTGTCGAAAATGCCGACTATGGCCGCG | This study |
gltA_R164L_rev | CGCGGCCATAGTCGGCATTTTCGACAGCAGGAGGAACGCGGCAAT | This study |
gltA_clon_for_1 | TGCGTAATACCGGCTAAAGGAGGAACCGTATGGCTGATACAAAAGCAAAACTC | This study |
gltA_clon_rev_1 | CATCCGCCAAAACAGAAGCTTTTAACGCTTGATATCGCTTTTAAAG | This study |
Plasmids | ||
pKD4 | amp | [53] |
pKD46 | amp | [53] |
pCP20 | amp cho | [53] |
pEXT20 | amp | [28] |
pACT3 | cho | [28] |
pACT3w-ppcwt | pACT3 expressing wild-type ppc gene from E. coli from weak RBS | This study |
pACT3w-ppcK620S | pACT3 expressing ppcK620S mutant gene from E. coli from weak RBS | This study |
pEXT20w-ppcK620S | pEXT20 expressing ppcK620S mutant gene from E. coli from strong RBS | This study |
pACT3s-ppcK620S | pACT3 expressing ppcK620S mutant gene from E. coli from strong RBS | This study |
pACT3w-ppcK620S-gltAR164L | pACT3w-ppcK620S expressing gltAR164L mutant gene from E. coli | This study |