Difference between revisions of "Part:BBa K4879003"

Revision as of 07:07, 11 October 2023

Yarrowia lipolytica HygR marker

Resistance marker gene for Hygromycin B, designed for expression 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 deletion in our chassis. In order to use the selection marker, we had to use 200µg/ml of hygromycin B in our selection plates.

Hygromycin B tends to kill cells by interfering in polypeptide synthesis, making the organism unable to synthesise proteins. The selection marker gene, which codes for a phosphotransferase, confers resistance by modifying the hygromycin B drug molecule; making it ineffective.

The protein structure of HygR, as predicted by AlphaFold

Design

The transcriptional unit for HygR (BBa_K4879009) is designed with the coding sequence flanked by the TEF1 promoter (BBa_K3629001) upstream and the XPR2 terminator (BBa_K3629004) downstream.

Sequence and Features

Assembly Compatibility:

10
INCOMPATIBLE WITH RFC[10]
Illegal EcoRI site found at 153
Illegal EcoRI site found at 247
12
INCOMPATIBLE WITH RFC[12]
Illegal EcoRI site found at 153
Illegal EcoRI site found at 247
21
INCOMPATIBLE WITH RFC[21]
Illegal EcoRI site found at 153
Illegal EcoRI site found at 247
23
INCOMPATIBLE WITH RFC[23]
Illegal EcoRI site found at 153
Illegal EcoRI site found at 247
25
INCOMPATIBLE WITH RFC[25]
Illegal EcoRI site found at 153
Illegal EcoRI site found at 247
1000
COMPATIBLE WITH RFC[1000]

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 The protein structure of HygR, as predicted by AlphaFold
+===Design===
+The transcriptional unit for HygR [https://parts.igem.org/Part:BBa_K4879009 (BBa_K4879009)] is designed with the coding sequence flanked by the TEF1 promoter [https://parts.igem.org/Part:BBa_K3629001 (BBa_K3629001)] upstream and the XPR2 terminator [https://parts.igem.org/Part:BBa_K3629004 (BBa_K3629004)] downstream.