Difference between revisions of "Part:BBa K3866032"

 
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This is the E. coli β-glucosidase gene bglX with N20 signal peptide instead of its native signal peptide.
 
This is the E. coli β-glucosidase gene bglX with N20 signal peptide instead of its native signal peptide.
  
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===Usage and Biology===
 
===Usage and Biology===
  
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Bglx belongs to the family of β-D-Glucosidases. These enzymes catalyze the hydrolysis of cellobiose (1,4-β-D-glucopyranosyl-D-glucopyranose) into two molecules of glucose. The bgIX gene is located adjacent to the dld gene at 47.8 min or 2225 kb on the E. coli chromosome. The presence of a 20 amino acids signal peptide (at the N-terminal) is consistent with the periplasmic location of the mature protein. The replacement of native signal peptide with N20- signal peptide is expected to make the mature protein an extracellular product.
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<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K3866032 SequenceAndFeatures</partinfo>
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===Design Notes===
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In order to construct this part, we replaced the native signal peptide with the N20 signal peptide sequence which is: 5’-atggaaggcaacacccgcgaagataactttaaacatctgctgggcaacgataacgtgaaacgcccgagcgaagcg-3’. The coding sequence was domesticated. We removed BsmBI, BsaI, BtgZI, BpiI sites in order to be compatible with GoldenBraid and MoClo. The sequence is present in pUPD2 BBa_K3505007 and has overhangs compatible for GoldenBraid cloning. The CDS has position B3-B5.
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===Verification of cloning===
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===Source===
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Synthesized by IDT.
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===Experimental Use and Experience===
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===Sequence and Features===
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<partinfo>BBa_K3866032 SequenceAndFeatures</partinfo>
  
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===References===
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===Functional Parameters===
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<partinfo>BBa_K3866032 parameters</partinfo>
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Revision as of 10:13, 27 September 2021


N20bglΧ GB compatible with B3-B5

This is the E. coli β-glucosidase gene bglX with N20 signal peptide instead of its native signal peptide.

Usage and Biology

Bglx belongs to the family of β-D-Glucosidases. These enzymes catalyze the hydrolysis of cellobiose (1,4-β-D-glucopyranosyl-D-glucopyranose) into two molecules of glucose. The bgIX gene is located adjacent to the dld gene at 47.8 min or 2225 kb on the E. coli chromosome. The presence of a 20 amino acids signal peptide (at the N-terminal) is consistent with the periplasmic location of the mature protein. The replacement of native signal peptide with N20- signal peptide is expected to make the mature protein an extracellular product.

Design Notes

In order to construct this part, we replaced the native signal peptide with the N20 signal peptide sequence which is: 5’-atggaaggcaacacccgcgaagataactttaaacatctgctgggcaacgataacgtgaaacgcccgagcgaagcg-3’. The coding sequence was domesticated. We removed BsmBI, BsaI, BtgZI, BpiI sites in order to be compatible with GoldenBraid and MoClo. The sequence is present in pUPD2 BBa_K3505007 and has overhangs compatible for GoldenBraid cloning. The CDS has position B3-B5.

Verification of cloning

Source

Synthesized by IDT.

Experimental Use and Experience

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 AgeI site found at 1637
    Illegal AgeI site found at 1859
    Illegal AgeI site found at 2048
  • 1000
    COMPATIBLE WITH RFC[1000]

References