Part:BBa_K1351009

57 aa flexible protein linker

Flexible linker of 57 amino acids for translational fusion of protein domains. Used in the [http://2014.igem.org/Team:LMU-Munich/Project/Bakillus BaKillus project] to display pathogen-specific peptides on the surface of B. subtilis with the YhcR/YhcS system to mediate adhesion of B. subtlis to pathogens.

The YhcR/YhcS systems consists of four components: the sortase substrate with a signal peptide (BBa_K1351008), a linker (this part) and a cell wall-anchoring domain (BBa_K1351010) as well as the sortase YhcS itself (BBa_K1351011), whose overexpression increases the efficiency of the surface display.

This part was generated in a modified version of RFC25, where a strong Shine Dalgarno Sequence (SD) is included, and has the following prefix and suffix:

prefix with EcoRI, NotI, XbaI, SD and NgoMIV:	GAATTCGCGGCCGCTTCTAGAGTAAGGAGGAGCCGGC
suffix with AgeI, SpeI, NotI and PstI:	ACCGGTTAATACTAGTAGCGGCCGCTGCAG

Sites of restriction enzymes generating compatible overhangs have the same color:

EcoRI and PstI in blue, NotI in green, XbaI and SpeI in red, NgoMIV and AgeI in orange. Shine-Dalgarno sequence and stop codons are underlined.

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
  INCOMPATIBLE WITH RFC[1000]

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