Difference between revisions of "Part:BBa K844012"

Latest revision as of 02:38, 4 October 2012

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. This part with 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 (BBa K844013) contains additional tRNAs for the glycine, proline, and serine amino acids, as well as two additional tRNAs that have 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

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 196
1000
COMPATIBLE WITH RFC[1000]

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 <partinfo>BBa_K844012 short</partinfo>
-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.
+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. This part with 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 (BBa K844013) contains additional tRNAs for the glycine, proline, and serine amino acids, as well as two additional tRNAs that have 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.
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