This region codes for the Acetyl-CoA Synthase in the acetogenic bacterium Acetobacterium woodii. To be more precise it codes for the acetyl-CoA synthase subunit (ACS) of the CODH/ACS-complex which catalyzes the formation of acetyl-CoA from CO and [CH3]-Co-FeS-P. This enzyme is crucial for the complementation of the Wood-Ljungdahl metabolic pathway. The gene coding fragment is flanked by specific restriction sites and overhangs. Upstream it is flanked by a Xho1 restriction site and a 5 base pairs overhang. Downstream it is flanked by a Nhe1 restriction site and a 5 base pairs overhang.
This region codes for the Acetyl-CoA Synthase in the acetogenic bacterium Clostridium ljungdahlii. To be more precise it codes for the acetyl-CoA synthase subunit (ACS) of the CODH/ACS-complex which catalyzes the formation of acetyl-CoA from CO and [CH3]-Co-FeS-P. This enzyme is crucial for the complementation of the Wood-Ljungdahl metabolic pathway. The gene coding fragment is flanked by specific restriction sites and overhangs. Upstream it is flanked by a Xho1 restriction site and a 5 base pairs overhang. Downstream it is flanked by a Nhe1 restriction site and a 5 base pairs overhang.
This is a fragment with a length of 4.942 kb, which was amplified out of genomic DNA of Acetobacterium woodii. It consists of the genes ascB1, cooC2 and acsA. cooC2 and acsA are immediately laying upstream of acsB1. acsB1 codes for the acetyl-CoA synthase subunit (ACS) of the CODH/ACS-complex which catalyzes the formation of acetyl-CoA from CO and [CH3]-Co-FeS-P. cooC2 codes for the putative CODH nickel-insertion accessory protein. acsA partially codes for the carbon monoxide dehydrogenase subunit of the CODH/ACS-complex, which catalyzes the reduction of CO2 to CO. All these genes are parts of the Wood-Ljungdahl pathway of acetogenic bacteria. The gene coding fragment is flanked by specific restriction sites and overhangs. Upstream it is flanked by a Sal1 restriction site and a 5 base pairs overhang. Downstream it is flanked by a Nhe1 restriction site and a 5 base pairs overhang.
This is a fragment with a length of 3.445 kb, which was synthesized by de-novo-synthesis with the genomic background of Acetobacterium woodii. It consists of the genes ascD, acsC and acsE. acsD and acsC code for the acetyl-CoA synthase subunit (ACS) of the CODH/ACS-complex which transfers a methyl-group from Co-FeS-P to the Ni-Fe complex. acsE codes for the methyltransferase subunit which transfers the methyl group generated from CO2 to Co-FeS-P. All these genes are parts of the Wood-Ljungdahl pathway of acetogenic bacteria. The gene coding fragment is flanked by specific restriction sites and overhangs. Upstream it is flanked by a Sal1 restriction site and a 5 base pairs overhang. Downstream it is flanked by a SfaA1 restriction site and a 5 base pairs overhang.
This is a fragment with a length of 4.12 kb, which was synthesized by de-novo-synthesis with the genomic background of Acetobacterium woodii. It consists of the genes cooC1, acsV, AWO_c10690 and AWO_c10700. CooC1 codes for an accessory protein of the Ni2+ insertion. acsV regenerates the corrinoid iron sulfur protein. AWO_c10690 and AWO_c10700 are putative proteins. All these genes are parts of the Wood-Ljungdahl pathway of acetogenic bacteria. The gene coding fragment is flanked by specific restriction sites and overhangs. Upstream it is flanked by a Sal1 restriction site and a 5 base pairs overhang. Downstream it is flanked by a Pac1 restriction site and a 5 base pairs overhang.
This is a fragment with a length of 1.573 kb, which was synthesized by de-novo-synthesis with the genomic background of Acetobacterium woodii. It consists of the genes metV and metF which encode two subunits of the methylene-THF reductase. All these genes are parts of the Wood-Ljungdahl pathway of acetogenic bacteria. The gene coding fragment is flanked by specific restriction sites and overhangs. Upstream it is flanked by a Pac1 restriction site and a 5 base pairs overhang. Downstream it is flanked by a Nhe1 restriction site and a 5 base pairs overhang.
This is a fragment with a length of 4.965 kb, which was amplified out of the genomic DNA of Acetobacterium woodii. It consists of the genes ascB1, cooC2 and acsA. cooC2 and acsA are immediately laying upstream of acsB1. acsB1 codes for the acetyl-CoA synthase subunit (ACS) of the CODH/ACS-complex which catalyzes the formation of acetyl-CoA from CO and [CH3]-Co-FeS-P. CooC2 codes for the putative CODH nickel-insertion accessory protein. acsA partially codes for the carbon monoxide dehydrogenase subunit of the CODH/ACS-complex, which catalyzes the reduction of CO2 to CO. All these genes are parts of the Wood-Ljungdahl pathway of acetogenic bacteria. The gene coding fragment is flanked by specific restriction sites and overhangs. Upstream it is flanked by a SfaA1 restriction site and a 5 base pairs overhang. Downstream it is flanked by a Nhe1 restriction site and a 5 base pairs overhang.
pMTL83151 is based on the modular plasmid system ClosTron (Heap et al., 2006) with the Gram-positive replicon pCB102 and ColE1 as a Gram-negative replicon. traJ facilitates the transfer of the plasmid and the catP cassette serves as an antibiotic (chloramphenicol) selection marker. lacZ codes for β-galactosidase, sits at the multiple cloning site and therefore serve as the transformation control.
This composite part consists of the plasmid backbone pMTL83151 (BBa_K4404007) and acsB2 (BBa_K4404001) which is under the control of the thiolase promoter (BBa_K2715019). The constitutive promoter from the C. acetobutylicum thiolase gene functions in Gram-negative (E. coli) and in Gram-positive (Clostridium saccharoperbutylacetonicum) bacteria. The insert acsB2 codes for the acetyl-CoA synthase subunit (ACS) of the CODH/ACS-complex which catalyzes the formation of acetyl-CoA from CO and [CH3]-Co-FeS-P. acsB2 was inserted into the backbone pMTL83151 to complement the Wood-Ljungdahl metabolic pathway for CO2 assimilation.
This composite part consists of plasmid backbone pMTL83151 (BBa_K4404007) and acsB1 (BBa_K4404000) which is under the control of the PbgaL promotor (BBa_K2992023). The promoter from C. perfringens functions in Gram-negative (E. coli) and in Gram-positive (Clostridium saccharoperbutylacetonicum) bacteria and is inducible by lactose. The insert acsB1 codes for the acetyl-CoA synthase subunit (ACS) of the CODH/ACS-complex which catalyzes the formation of acetyl-CoA from CO and [CH3]-Co-FeS-P. acsB1 was inserted into the backbone pMTL83151 to complement the Wood-Ljungdahl metabolic pathway for CO2 assimilation.
This composite part consists of plasmid backbone pMTL83151 (BBa_K4404007) and acsB2 (BBa_K4404001) which is under the control of the PbgaL promoter (BBa_K2992023). The promoter from C. perfringens functions in Gram-negative (E. coli) and in Gram-positive (Clostridium saccharoperbutylacetonicum) bacteria and is inducible by lactose. The insert acsB2 codes for the acetyl-CoA synthase subunit (ACS) of the CODH/ACS-complex which catalyzes the formation of acetyl-CoA from CO and [CH3]-Co-FeS-P. acsB2 was inserted into the backbone pMTL83151 to complement the Wood-Ljungdahl metabolic pathway for CO2 assimilation.