Part:BBa_K2974101

Medium Promoter (BBa_J23106) Toehold GFP

BBa_J23106 Toehold eGFP is a construct designed to be utilized as a riboregulatory biosensor in conjunction with part BBa_K2550001. The part was assembled using a De-Novo Toehold Switch construct originally designed in the Collins Lab at the Massachuchets Institute of Technology and a smedium-strength promoter BBa_J23106 of the Anderson Promoter family. Promoter BBa_J23106 is from the same parts collection as BBa_J23100 and was utilized to reduce overexpression due to diminished strength. Composite part BBa_K2974500 provides both a proof of concept for part BBa_K2974310 and characterization for promoter expression within the T7 promoter Toehold Ribosome Switch with the eGFP reporter expression system.

Description
Toehold switches are synthetic RNA strands that mimic messenger RNA sequences. They contain a complementary recognition sequence for a specific sequence of RNA stimuli , and a ribosomal binding site where a ribosome binds to initiate the translation of a reporter protein. The switch has a hairpin loop structure that is formed through binding to complementary sections of its own sequence. When in the presence of the complementary trigger sequence the hairpin loop opens to allow downstream expression. The Ribosomal Binding Site and starting sequence are concealed in the toehold switch until initiated. Switches can provide rapid, convenient, in-field detection that can be developed in both cellular systems and cell-free tests.

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
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 728