Difference between revisions of "Part:BBa K863101:Design"
(→Design Notes) |
(→Design Notes) |
||
Line 6: | Line 6: | ||
===Design Notes=== | ===Design Notes=== | ||
− | The sequence starts with the rest of the Freiburg-prefix (ATG and ''Ngo''MIV-restriction-site). The A from the startcodon ATG is the last base of the ''Xba''I restriction-site of the | + | The sequence starts with the rest of the Freiburg-prefix (ATG and ''Ngo''MIV-restriction-site). The A from the startcodon ATG is the last base of the ''Xba''I restriction-site of the [https://parts.igem.org/Assembly_standard_25 Standard 25 Prefix (Read here)]. I isolated 12 conserved Bases (4 AS) upstream of the cellulose binding domain and 9 bases downstream of the exoglucanase gene and added a short N-terminal (Glycine, Serine) linker. The sequence ends with the ''Age''I-restriction-site of the Freiburg-Suffix, to make in frame assembly possible. |
Primers that were used to make this BioBrick: | Primers that were used to make this BioBrick: |
Revision as of 19:34, 26 September 2012
Cellulose binding Domain of Cellulomonas Fimi Exoglucanse (Freiburg-Standard)
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 4
Illegal AgeI site found at 337 - 1000COMPATIBLE WITH RFC[1000]
Design Notes
The sequence starts with the rest of the Freiburg-prefix (ATG and NgoMIV-restriction-site). The A from the startcodon ATG is the last base of the XbaI restriction-site of the Standard 25 Prefix (Read here). I isolated 12 conserved Bases (4 AS) upstream of the cellulose binding domain and 9 bases downstream of the exoglucanase gene and added a short N-terminal (Glycine, Serine) linker. The sequence ends with the AgeI-restriction-site of the Freiburg-Suffix, to make in frame assembly possible.
Primers that were used to make this BioBrick:
CBDcex_Freiburg-Prefix | GCTAGAATTCGCGGCCGCTTCTAGATGGCCGGCGGTCCGGCCGGGTGCCAGGTG |
CBDcex_2AS-Link_Frei | CTGCAGCGGCCGCTACTAGTATTAACCGGTGCTGCCGCCGACCGTGCAGGGCGTGC |
Source
Cloning-Vector with a CBDcex-Barnase fusion-protein