Coding

Part:BBa_K3394000

Designed by: Talia Shaler   Group: iGEM20_BGU-Israel   (2020-10-05)
Revision as of 09:15, 5 October 2020 by Talia sh (Talk | contribs)


Coding Sequence of Endo5a cellulase (for E. coli)

This part consists the coding region of Endo5A, an endo-1,4-beta-glucanases. Endo5a breaks down cellulose by making internal cuts in the cellulose chain. This enzyme is part of the complete cellulose hydrolysis reaction which involves several enzymes: endo-1,4-beta-glucanases, exo-1,4-beta-glucanases (or cellobiohydrolases) and beta-glucosidases.

It is a relatively small protein with an optimal temperature of 50 Celsius degrees and an optimal pH range of 6-7. The NCBI Genbank number for Endo5a is HQ657203.1 (DNA), AEB00655.1 (amino acid). This construct was synthesized by IDT for use in Escherichia coli.

We verified that the sequence is optimized for use in Escherichia coli. Furthermore, Endo5a was expressed with an N-terminal histidine tag (6-histidine tag) instead of the 17 first amino acids that served as a signal peptide (51 nucleotides).

Endo5A was isolated from the bacterial flora found in the gut of a cotton bollworm (Helicoverpa armigera) which secretes a variety of plant-hydrolyzing enzymes.


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 898
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 898
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 898
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 898
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 898
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
None