Part:BBa_K4339000

Non-polar amino acid transporter (CycA)

CycA is an inner membrane permease [1], which can facilitate the uptake of non-polar amino acids (including D-alanine, D-serine, glycine and L-alanine) via a proton symport mechanism [2]. CycA was first identified in the K12 strain of E. coli.

CycA was codon-optimised for expression in E. coli BL21 DE3 strains and their derivatives. CycA was fused with the constitutive promoter PJ23100 and a C-terminus His tag and engineered into the strain BL21 DE3 pLysS. CycA was engineered into this strain with an aim to increase uptake of L-alanine from an alanine-doped LB growth medium, to facilitate synthesis of alanine-rich proteins such as MaSps (Major Ampullate Silk proteins).

Usage and Biology

Sequence and Features

The sequence originates from E. coli MG1655 K-12 (Hook et al. 2022)

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 340
  
• 1000
  COMPATIBLE WITH RFC[1000]

References

[1] - Hook C et al. The Escherichia coli Amino Acid Uptake Protein CycA: Regulation of Its Synthesis and Practical Application in l-Isoleucine Production. Microorganisms. 2022;10(3):647. doi: https://doi.org/10.3390/microorganisms10030647

[2] - CycA. ECOCYC version 26.0. Available at: https://biocyc.org/gene?orgid=ECOLI&id=EG12504 [Accessed 14/9/22]