Part:BBa_K1983015

PheP and tRNA-Phe under constitutive promoters

Overview

This biobrick part contains two coding elements: (BBa_K1983014): PheP, which is labeled with N-6XHis-Tag and put under a constitutive promoter (BBa_K823005), high strength RBS 9BBa_J61132) and terminator (BBa_B0017). A basic biobrick part (BBa_K1983011): tRNA-Phe (pheU) under constitutive promoter and terminatorPheP (Phenylalanine-specific permease) is a natural phenylalanine membrane transporter of the bacterium Escherichia coli.

PheP is a single integral membrane transporter which selectively transports L-phenylalanine and L-tyrosine as an antiporter using proton motive force. The activity of this transporter under natural expression is known to be 9 and 17,5 nmol/mgDW(cells) for L-phenylalanine and L-tyrosine respectively [1].

pheU gene from Escherichia coli encodes a tRNA transporting L-phenylalanine and is under an upstream constitutive promoter and a terminator found after the tRNA gene.

Cloning

This part was created by fusing a fragment received from IDT that contains a part (BBa_K1983014) with a biobrick part (BBa_K1983011) inside a pSB1C3 backbone. The received sequences were amplified using Uni-FW/RV primers and digested with EcoRI and SpeI. The fragments were cloned into pSB1C3 vector containing (BBa_K1983011) digested with EcoRI and XbaI.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 7
  Illegal NheI site found at 30
  Illegal NheI site found at 98
  Illegal NheI site found at 1669
  Illegal NheI site found at 1692
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]