Difference between revisions of "Part:BBa K801073"

(→‎Cloning into pSB1C3)
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Yeast expression cassette for CaXMT1 ([https://parts.igem.org/Part:BBa_K801070 BBa_K801070]) controlled by the yeast TEF2 promoter ([https://parts.igem.org/Part:BBa_K801010 BBa_K801010]) and the yeast ADH1 terminator ([https://parts.igem.org/Part:BBa_K801012 BBa_K801012]).
 
Yeast expression cassette for CaXMT1 ([https://parts.igem.org/Part:BBa_K801070 BBa_K801070]) controlled by the yeast TEF2 promoter ([https://parts.igem.org/Part:BBa_K801010 BBa_K801010]) and the yeast ADH1 terminator ([https://parts.igem.org/Part:BBa_K801012 BBa_K801012]).
  
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This BioBrick is the generator for the enzyme xanthosine N-methyltransferase 1 (CaXMT1) and part of the caffeine synthesis pathway. For creating an expression cassette with all three enzymes of the caffeine synthesis pathway based on the substrate Xanthosine [https://parts.igem.org/Part:BBa_K801077 BBa_K801077] different promoters and terminators were assembled to each enzyme. CaXMT1 is regulated by the constitutive promoter Tef2, which is a strong yeast promoter. The used terminator Adh1, is a widely used yeast terminator. The Tef2 promoter was prefered compared to the Tef1 promoter (which is even stronger) in order to limit metabolic stress, which could result in a positive selection of natural mutants (with regard to genome integration).
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This BioBrick is the generator for the enzyme xanthosine N-methyltransferase 1 (CaXMT1) and part of the caffeine synthesis pathway. For creating an expression cassette with all three enzymes of the caffeine synthesis pathway [https://parts.igem.org/Part:BBa_K801077 BBa_K801077] based on the substrate Xanthosine different promoters and terminators were assembled to each enzyme. CaXMT1 is regulated by the constitutive promoter Tef2, which is a strong yeast promoter. The used terminator Adh1, is a widely used yeast terminator. The Tef2 promoter was prefered compared to the Tef1 promoter (which is even stronger) in order to limit metabolic stress, which could result in a positive selection of natural mutants (with regard to genome integration).
  
  

Revision as of 12:23, 3 October 2012

CaXMT1 expression cassette for yeast

Yeast expression cassette for CaXMT1 (BBa_K801070) controlled by the yeast TEF2 promoter (BBa_K801010) and the yeast ADH1 terminator (BBa_K801012).

This BioBrick is the generator for the enzyme xanthosine N-methyltransferase 1 (CaXMT1) and part of the caffeine synthesis pathway. For creating an expression cassette with all three enzymes of the caffeine synthesis pathway BBa_K801077 based on the substrate Xanthosine different promoters and terminators were assembled to each enzyme. CaXMT1 is regulated by the constitutive promoter Tef2, which is a strong yeast promoter. The used terminator Adh1, is a widely used yeast terminator. The Tef2 promoter was prefered compared to the Tef1 promoter (which is even stronger) in order to limit metabolic stress, which could result in a positive selection of natural mutants (with regard to genome integration).


Cloning into pSB1C3

The cloning into pSB1C3 was proved by performing an analytical digest.

TUM12 analyticaldigestCaXMT1cassette.jpg

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 138
  • 1000
    COMPATIBLE WITH RFC[1000]