Part:BBa_K844012
tRNA expression cassette for spider silk "F" proteins
The tRNA sequences for use in overexpression of spider silk proteins were selected from the Genomic tRNA Database for E. coli K-12. tRNA sequences with anti-codons recognizing the codons used in the optimized “B” and “F” subunits were selected, and their native sequences used. The six tRNAs (BBa K844012) corresponding to the codons used in the “F” construct were made into a single construct with each gene proceeded by the ZH14 12x tRNA promoter and followed by the downstream region of the E.coli Ala (GCC) sequence. The five tRNA construct contains additional tRNAs for the glycine, proline, and serine amino acids, as well as two additional tRNAs that has anticodons matching the threonine and arginine amino acids that is generated in the scar sites from use of the BioBrick Assembly Standard #23 cloning method. Note: All tRNAs are from E. coli K12 except the Arg(TCT) tRNA is from Escherichia_coli_O157H7 since the K12 ARG tRNA had an internal PstI site.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 196
- 1000COMPATIBLE WITH RFC[1000]
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