Coding

Part:BBa_K4620015

Designed by: Jānis Edmunds Daugavietis   Group: iGEM23_Latvia-Riga   (2023-10-12)
Revision as of 15:10, 12 October 2023 by Kristine11 (Talk | contribs)

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Skr-BURP tryptic peptide containing the core peptide

ADNVLFYPSYAAR is a peptide formed after tryptic digestion of Skr-BURP protein. It contains core peptide fragment VLFYPSY. VLFYPSY is a peptide that can be cyclised by BURP domain protein from African clubmoss (Selaginella kraussiana) (BBa_K4620000), forming selanine A. This side chain macrocyclisation reaction is catalysed by copper (II) ions. Cyclisation happens between the OH-group of tyrosine and Cβ of leucine or Cβ of tyrosine. A bicyclic compound is formed. As of this day, mechanism of this reaction has not been fully characterised.

In the nature, ADNVLFYPSYAAR is conjugated to the N-terminal domain of Skr-BURP. iGEM 2023 team Latvia-Riga produced this peptide separately in order to make trans-cyclisation reaction where the cyclisation substrate is not attached to the catalytic domain. We chose substrate a little bit longer than the core domain in order to make expression and purification easier. In the nature, ADNVLFYPSYAAR is conjugated to the N-terminal domain of Skr-BURP.

iGEM 2023 team Latvia-Riga produced this peptide separately in order to make trans-cyclisation reaction where the cyclisation substrate is not attached to the catalytic domain. We chose substrate a little bit longer than the core domain in order to make expression and purification easier.

Cloning

Our team cloned ADNVLFYPSYAAR gene into pEXP-GB1 expression vector, containing GB1 (domain B1 of Immunoglobulin G-binding protein G from Streptococcus sp.), N-terminal 8xHis tag and TEV protease cleavage site.


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
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
None