Difference between revisions of "Team:Tuebingen/Parts"

This part contains a 6-phosphofructoskinase from lactobacillus Johnsonii ATCC33200 that metabolizes fructose-6-phosphate to fructose-1,6-bisphosphate. This part is RFC 10 compatible. The part's codon usage is optimized for expression in lactobacilli. The codon usage of this part was optimized to balance DNA synthesis and expression in L Johnsonii. Therefore longterm and shortterm repeats had to be modified and biobrick restriction sites were removed. The part was modified for RFC10 at the prefix and suffix of the part.

BBa_K1882005

This part encodes BBa_K1882002 and BBa_K1882000 resulting in a l.johnsonii ATCC33200 optimized phosphofructokinase under the control of a constitutive LDH-promotor. This leads to overexpression of the phosphofructokinase in lactobacillus Johnsonii ATCC33200 when transferring this part into a lactobacillus expression vector.

BBa_K1882001

This part contains a the promoter of the Lactobacillus Johnsonii ATCC33200 lactate dehydrogenase, a constitutively expressed protein. This part is RFC10 compatible

Fructose Transporter

This part encodes a carbohydrate transporter from lactobacillus Johnsonii ATCC 33200 that imports fructose into the target cell. In this process, fructose is phosphorylated by a N-phosphohistidine residue to yield fructose-6-phosphate and histidine. The protein is part of the ABC transporter superfamily. This part ist RFC 10 compatible.The part's codon usage is optimized for expression in lactobacilli. The codon usage of this part was optimized to balance DNA synthesis and expression in l.Johnsonii. Therefore longterm and shortterm repeats had to modified and biobrick restriction sites were removed.The part was modified for RFC10 at the prefix and suffix of the part.

BBa_K1882006

This part contains a the promoter of the Lactobacillus Johnsonii ATCC33200 lactate dehydrogenase, a constitutively expressed protein. This part is RFC10 compatible

BBa_K1882007

This part encodes a Ketohexokinase (ATP:D-fructose 1-phosphotransferase) from rattus norvegicus optimized for expression in saccharomyces cerevisiae. The ketohexokinase converts fructose to fructose-1-phosphate using ATP to perform the phosphorylation. This protein is naturally not common in yeast and functions as toxin of the cell by trapping phosphate and therefore ATP in fructose-1-phosphate, not being able to be further metabolized. This result in no further growth of transformed yeast in fructose containing media (Donaldson et al. 1993). This part is RFC25 and RFC10 compatible.

BBa_K1882000

This part encodes for BBa_J23102 with an ApaI restriction site upstream of it. In combination with BBa_K1882010 enables this part to clone a construct in a RFC compatible plasmid.

BBa_K1882009

This part encodes for the rrnB T1 Terminator (BBa_B0010) followed by a T7 Terminator (BBa_B0012) and BamHI and HindIII restriction sites. In combination with BBa_K1882009 enables this part to clone a construct in a RFC compatible plasmid.

BBa_K1882010

This part encodes for a codon optimized vesion of UDP-N-acetylglucosamine kinase (PezT toxin). Uppon expression produces the Toxin UDP-N-acetyl-alpha-D-glucosamine 3'-phosphate which inhibits MurA and therefore cellwallsynthesis in ecoli as well as loctobacillus johnsonii. Besides the sequence for the PezT toxin contains this part an upstream located Ribosom Binding Site and could therefore cloned direct behind the Promotor of choice.

BBa_K1882011

This part contains the zinc finger ZF ZFN-L, see also Lombardo et al., 2007. The part is compatible with RFC10.

BBa_K1882012

This part contains the zinc finger ZFK230R, see also Lee et al., 2010. The part is compatible with RFC10.

BBa_K1882013

This part contains the zinc finger ZF-C7C7, see also Liu et al., 1997. The part is compatible with RFC10.

BBa_K1882014

This part is a composite part of the following components: Apa-pRha + RBS-PezT + Terminator-Bam-Hind. It is RFC10 compatible, and can be used for negative selection.

BBa_K1882015