Composite

Part:BBa_K4275036

Designed by: Yujun Li   Group: iGEM22_GreatBay_SCIE   (2022-10-11)
Revision as of 01:01, 12 October 2022 by Jerryatcg (Talk | contribs)


pLAC4-KmarxMFα-NpaBGS-t-tTDH1

The composite part contains a pLac4 promoter, Kluyveromyces marxianus mating factor alpha, beta-glucosidase NpaBGS fused with type I dockerin and tTDH1 terminator. NpaBGS is a cellulase active on oligosaccharides(short polysaccharides) and cellobiose(disaccharide) that cleaves glycosidic bonds. The Kluyveromyces marxianus mating factor alpha is fused in order to enable secretion of NpaBGS by host K.marxianus.


Usage and Biology

Kluyveromyces marxianus mating factor alpha fused with NpaBGS would lead peptide of NpaBGS-t through ER and Golgi apparatus and be secreted. NpaBGS is a β-glucosidase containing two glycosyl hydrolase Family 3 domains at N and C terminals .It was discovered from the buffalo rumen fungus Neocallimastix patriciarum W5. It’s activity could be enhanced at 50°C by adding Mg2+ or Mn2+ ions.


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 331
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 331
    Illegal NheI site found at 89
    Illegal NheI site found at 1783
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 331
    Illegal XhoI site found at 1252
    Illegal XhoI site found at 2858
    Illegal XhoI site found at 3937
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 331
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 331
    Illegal NgoMIV site found at 2355
    Illegal AgeI site found at 1369
    Illegal AgeI site found at 1387
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
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