Difference between revisions of "Part:BBa K4879004"
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===Usage and Biology=== | ===Usage and Biology=== | ||
+ | Due to the limited number of auxotrophic markers available for <i>Yarrowia lipolytica</i>, we had to use antibiotic-resistance markers for the selection of possible attempted deletions in our chassis. In order to use the selection marker, we had to use 1000-2000µg/ml of mycophenolic acid (MPA) in our selection plates. | ||
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+ | MPA tends to kill cells by inhibiting inosine monophosphate dehydrogenase (IMPDH), an important enzyme in the de novo biosynthesis of GMP. The selection marker gene, which codes for an IMPDH variant, confers resistance by bypassing this inhibition. | ||
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+ | <html><img src="https://static.igem.wiki/teams/4879/wiki/bba-k4879004-guab.png"width="250"height="250"></html> | ||
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+ | The protein structure of JcFATA, as predicted by AlphaFold | ||
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Revision as of 06:24, 11 October 2023
Yarrowia lipolytica guaB marker
Mycophenolic acid resistance marker, codon optimized for Yarrowia lipolytica
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Usage and Biology
Due to the limited number of auxotrophic markers available for Yarrowia lipolytica, we had to use antibiotic-resistance markers for the selection of possible attempted deletions in our chassis. In order to use the selection marker, we had to use 1000-2000µg/ml of mycophenolic acid (MPA) in our selection plates.
MPA tends to kill cells by inhibiting inosine monophosphate dehydrogenase (IMPDH), an important enzyme in the de novo biosynthesis of GMP. The selection marker gene, which codes for an IMPDH variant, confers resistance by bypassing this inhibition.
The protein structure of JcFATA, as predicted by AlphaFold
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 565
Illegal EcoRI site found at 1019
Illegal PstI site found at 878 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 565
Illegal EcoRI site found at 1019
Illegal PstI site found at 878 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 565
Illegal EcoRI site found at 1019
Illegal BamHI site found at 1081
Illegal XhoI site found at 851 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 565
Illegal EcoRI site found at 1019
Illegal PstI site found at 878 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 565
Illegal EcoRI site found at 1019
Illegal PstI site found at 878
Illegal NgoMIV site found at 868
Illegal NgoMIV site found at 1093 - 1000COMPATIBLE WITH RFC[1000]