Collections/Cell-Free TX-TL

This collection is a user contributed collection, and is not under curation by iGEM HQ/Registry.

Next vivo

Our part collection will contain all the major biomolecules required for a cell-free transcription-translation system providing them in an easy to express and purify way that anyone can easily produce for their experimental purposes. These biomolecules include:

  • 38 Transcription and Translation (TX-TL) protein genes codon optimized for E. coli expression
  • 20 Next vivo MS2-tagged tRNA isolation genes
  • 2 MS2-tagged ribosomal rRNA genes for isolation of functional ribosomes from the cell lysate


TX-TL Proteins: The initial focus for this collection was submitting all 38 TX-TL protein genes, codon optimized, hexahistidine tagged, and under the control of a T7 promoter system for controlled expression. We have submitted and characterized 9 of these parts to the registry in pSB1C3, characterized expression of 17 parts, and are working toward completing the remaining parts following the competition.

tRNA: The tRNA component of the Next vivo system was another of our focuses for this years project. Currently, no simple protocol exists for efficient and specific isolation of tRNA from the cell. We are happy to announce that we were able to develop a system that allows for the purification of specific, fully modified tRNA from cell lysate. The first tRNA in our part collection is that of tRNAPhe. With the success of our purification strategy, we plan to synthesize the remaining tRNA in our expression construct for submission into this collection.

Ribosomes: Currently we plan to BioBrick both the 23S and 16S E. coli ribosomal rRNA genes with the MS2 hairpin added into either sequence at a region that places it externally on the 3D ribosome structure. This design has previously been used before by another research lab to purify whole ribosomes using the MS2 binding protein tagged with a hexahistidine tag.

MS2BP: The MS2 binding protein is a central component to our tRNA and ribosome purification strategies. It was previously submitted by the 2016 Lethbridge iGEM team and we feel it would be a great addition to our Next vivo Part Collection.

Validation: Finally, we have also included our measurement control construct for the complete TX-TL system. This part expresses a RNA Spinach tagged EYFP mRNA and EYFP protein, that in the addition of DFHBI produces both green and yellow fluorescence indicating transcription and translation respectively.


More...
NameDescriptionLengthCreated byDocumentationUsesTypeStatus
BBa_K2109108MS2 6XHis (V29I/d1FG)640Andy Hudson4145 CompositeIt's complicated
BBa_K2443000AlaRS optimized for expression in E. coli 2862Lethbridge iGEM 20171602 CompositeNot in stock
BBa_K2443001ArgRS optimized for expression in E. coli 1965Lethbridge iGEM 20172060 CompositeIt's complicated
BBa_K2443002AsnRS optimized for expression in E. coli 1632Lethbridge iGEM 20171608 CompositeNot in stock
BBa_K2443003AspRS optimized for expression in E. coli 2004Lethbridge iGEM 20172105 CompositeIt's complicated
BBa_K2443004CysRS optimized for expression in E. coli 1617Lethbridge iGEM 20171781 CompositeIt's complicated
BBa_K2443005GlnRS optimized for expression in E. coli 1899Lethbridge iGEM 20171605 CompositeIt's complicated
BBa_K2443006GluRS optimized for expression in E. coli 1650Lethbridge iGEM 20171614 CompositeNot in stock
BBa_K2443007GlyRS Alpha subunit optimized for expression in E. coli 1143Lethbridge iGEM 20172087 CompositeNot in stock
BBa_K2443008GlyRS Beta subunit optimized for expression in E. coli 2301Lethbridge iGEM 20172075 CompositeIt's complicated
BBa_K2443009HisRS optimized for expression in E. coli 1506Lethbridge iGEM 20172070 CompositeNot in stock
BBa_K2443010IleRS optimized for expression in E. coli 3048Lethbridge iGEM 20171608 CompositeNot in stock
BBa_K2443011LeuRS optimized for expression in E. coli 2814Lethbridge iGEM 20171602 CompositeNot in stock
BBa_K2443012LysRS optimized for expression in E. coli 1750Lethbridge iGEM 20171769 CompositeIt's complicated
BBa_K2443013MetRS optimized for expression in E. coli 2265Lethbridge iGEM 20172402 CompositeIt's complicated
BBa_K2443014PheRS Alpha subunit optimized for expression in E. coli 1215Lethbridge iGEM 20172309 CompositeIt's complicated
BBa_K2443015PheRS Beta subunit optimized for expression in E. coli 2619Lethbridge iGEM 20171614 CompositeNot in stock
BBa_K2443016ProRS optimized for expression in E. coli 1950Lethbridge iGEM 20171602 CompositeNot in stock
BBa_K2443017SerRS optimized for expression in E. coli 1524Lethbridge iGEM 20172299 CompositeIt's complicated
BBa_K2443018ThrRS optimized for expression in E. coli 2160Lethbridge iGEM 20171606 CompositeNot in stock
BBa_K2443019TrpRS optimized for expression in E. coli 1236Lethbridge iGEM 20172224 CompositeIt's complicated
BBa_K2443020TyrRS optimized for expression in E. coli 1506Lethbridge iGEM 20171604 CompositeNot in stock
BBa_K2443021ValRS optimized for expression in E. coli 3087Lethbridge iGEM 20171773 CompositeIt's complicated
BBa_K2443022MTF optimized for expression in E. coli 1179Lethbridge iGEM 20172063 CompositeNot in stock
BBa_K2443023IF1 optimized for expression in E. coli 456Lethbridge iGEM 20171679 CompositeIt's complicated
BBa_K2443024IF2 optimized for expression in E. coli 2904Lethbridge iGEM 20171510 CompositeIt's complicated
BBa_K2443025IF3 optimized for expression in E. coli 774Lethbridge iGEM 20171683 CompositeIt's complicated
BBa_K2443026EF-G optimized for expression in E. coli 2346Lethbridge iGEM 20171647 CompositeIt's complicated
BBa_K2443027EF-Tu optimized for expression in E. coli 1419Lethbridge iGEM 20173149 CompositeIt's complicated
BBa_K2443028EF-Ts optimized for expression in E. coli 1083Lethbridge iGEM 20173070 CompositeIt's complicated
BBa_K2443029RF1 optimized for expression in E. coli 1314Lethbridge iGEM 20171794 CompositeIt's complicated
BBa_K2443030RF2 optimized for expression in E. coli 1329Lethbridge iGEM 20171787 CompositeIt's complicated
BBa_K2443031RF3 optimized for expression in E. coli 1821Lethbridge iGEM 20172090 CompositeIt's complicated
BBa_K2443032RRF optimized for expression in E. coli 789Lethbridge iGEM 20172112 CompositeNot in stock
BBa_K2443033MK optimized for expression in E. coli 879Lethbridge iGEM 20172433 CompositeNot in stock
BBa_K2443034CK optimized for expression in E. coli 1377Lethbridge iGEM 20172078 CompositeIt's complicated
BBa_K2443035NDK optimized for expression in E. coli 717Lethbridge iGEM 20171883 CompositeIt's complicated
BBa_K2443036PPiase optimized for expression in E. coli 822Lethbridge iGEM 20171710 CompositeIt's complicated
BBa_K2443037T7 RNA Polymerase optimized for expression in E. coli 2886Lethbridge iGEM 20172246 CompositeNot in stock
BBa_K2443038tRNAPhe-MS2300Lethbridge iGEM 20173980 CompositeNot in stock
BBa_K2443039EYFP-Spinach transcription and translation (TX-TL) measurement device1063Lethbridge iGEM 20171503 MeasurementNot in stock
BBa_K2722000Exonuclease V, Subunit RecC3369Enikö Baligács3320 CodingIt's complicated
BBa_K2722001Exonuclease V, Subunit RecD1827Enikö Baligács3261 CodingIt's complicated
BBa_K27220026 times Chi recombination hotspot to inhibit Exonuclease V (RecBCD) activity92Enikö Baligács4459 DNAIt's complicated
  • To add to this list, please use the category //collections/cell-free_tx-tl