Part:BBa_K3102043:Design
Lactate production (ldhA, lldP, ACS operon)
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1478
Illegal XhoI site found at 3404 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 489
Illegal AgeI site found at 1614
Illegal AgeI site found at 2077
Illegal AgeI site found at 2515
Illegal AgeI site found at 2674
Illegal AgeI site found at 2686
Illegal AgeI site found at 3833 - 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 219
Illegal SapI.rc site found at 6698
Design Notes
In ACS operon, we mutated a Cytosine into Thymine at the 1563bp position (C1563T) because of the Esp3I enzyme that we used for our Golden Gate Assembly (GGA) construct. We also mutated an Adenine into Guanine and a Cytosine in Thymine at the 2425bp and 2428bp position (A2425G & C2428T), respectively, because of the EcoRI enzyme. (BBa_K3102013)
Source
LDHA, E.coli, sequence found through Ebi https://www.ebi.ac.uk/ena/data/view/AAC74462
LLDP, E.coli, sequence found through Trypanocyc http://vm-trypanocyc.toulouse.inra.fr/ECOLI/sequence?type=GENE&object=EG11961
ACS operon, sequence comes from E.coli and was found through Biocyc: https://biocyc.org/ECOLI/sequence-delimited?chromosome=COLI-K12&object=EG11448&up=0&down=2160&right=4287371&left=4283253&type=true
Promoter (BBa_I719005), RBS (BBa_B0034) and Terminator (BBa_B0015) are from the iGEM Registry.
References
Bunch PK, Mat-Jan F, Lee N, Clark DP. "The ldhA gene encoding the fermentative lactate dehydrogenase of Escherichia coli". Microbiology. 1997;143(1 997):187–95.
Dong JM et al., Three overlapping lct genes involved in L-lactate utilization by Escherichia coli., Journal of Bacteriology, 1993 Oct;175(20):6671-8.
Gimenez R et al., The Gene yjcG, Cotranscribed with the Gene acs, Encodes an Acetate Permease in Escherichia coli, Journal of Bacteriology, 2003 Nov; 185(21):6448-55.
Nunez MF et al., Transport of L-Lactate, D-Lactate, and glycolate by the LldP and GlcA membrane carriers of Escherichia coli, Biochem Biophys Res Commun. 2002 Jan 18;290(2):824-9.