Difference between revisions of "Part:BBa K4879004"
 
(One intermediate revision by one other user not shown)
Line 15: Line 15:
  
  
−
The protein structure of JcFATA, as predicted by AlphaFold
+
The protein structure of IMPDH variant, as predicted by AlphaFold
 +
 
 +
===Design===
 +
The transcriptional unit for the guaB marker [https://parts.igem.org/Part:BBa_K4879010 (BBa_K4879010)] is designed with the coding sequence flanked by the TEF1 promoter [https://parts.igem.org/Part:BBa_K2983053 (BBa_K2983053)] upstream and the XPR2 terminator [https://parts.igem.org/Part:BBa_K3629004 (BBa_K3629004)] downstream.
  
  

Latest revision as of 12:06, 12 October 2023


Yarrowia lipolytica guaB marker

Mycophenolic acid resistance marker, codon optimized for Yarrowia lipolytica

-

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 IMPDH variant, as predicted by AlphaFold

Design

The transcriptional unit for the guaB marker (BBa_K4879010) is designed with the coding sequence flanked by the TEF1 promoter (BBa_K2983053) upstream and the XPR2 terminator (BBa_K3629004) downstream.


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 565
    Illegal EcoRI site found at 1019
    Illegal PstI site found at 878
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 565
    Illegal EcoRI site found at 1019
    Illegal PstI site found at 878
  • 21
    INCOMPATIBLE 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
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 565
    Illegal EcoRI site found at 1019
    Illegal PstI site found at 878
  • 25
    INCOMPATIBLE 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
  • 1000
    COMPATIBLE WITH RFC[1000]