Part:BBa_K4171016

PT7-tRNA(UTuX)

Backgrounds

This is the DNA sequence of native tRNA^UTuX (BBa_K4171009) of E.coli , which can bind with serine (Ser) by the help of enzyme SerRS (BBa_K4171001). With T7 promoter sequence (BBa_I719005) in the front, RNA polymerase is able to work properly on the template when doing in vitro RNA synthesis.

Fig. 1. In vitro Sec synthesis pathway

Usage

This sequence will be synthesized through in vitro RNA synthesis, which produces tRNA^UTuX transcript, the substrate of SerRS for seryl-tRNA aminoacylation in tRNA selenocysteine synthesis pathway.

Characterization

Fig. 2. Confirmation of P_T7 -tRNA^UTuX (BBa_K4171016) constructed through PCR. M: Marker; Lane1: P_T7 -tRNA^UTuX (114 bp)

Reference

[1]Fu, X., Crnković, A., Sevostyanova, A. and Söll, D., 2018. Designing seryl‐ tRNA synthetase for improved serylation of selenocysteine tRNA s. FEBS Letters, 592(22), pp.3759-3768.

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 71
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 61