Difference between revisions of "Part:BBa K525730:Design"
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===Design Notes=== | ===Design Notes=== | ||
*This is the complete sequence of NAD<sup>+</sup> -dependent DNA ligase with T7-promoter and RBS. | *This is the complete sequence of NAD<sup>+</sup> -dependent DNA ligase with T7-promoter and RBS. | ||
| − | *It is designed in [https://parts.igem.org/Assembly_standard_25 RFC 25] | + | *It is designed in [https://parts.igem.org/Assembly_standard_25 RFC 25] assembly standard |
| + | **AgeI restriction site upstream of stop codon to easily fuse a C-terminal domain to the coding sequence | ||
===Source=== | ===Source=== | ||
Latest revision as of 21:57, 21 September 2011
DNA ligase from Escherichia coli (LigA) with PT7 and RBS
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1449
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 2047
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 1663
Design Notes
- This is the complete sequence of NAD+ -dependent DNA ligase with T7-promoter and RBS.
- It is designed in RFC 25 assembly standard
- AgeI restriction site upstream of stop codon to easily fuse a C-terminal domain to the coding sequence
Source
- PCR on the Escherichia coli TOP 10 DNA cloned into BioBrick Vectors
- Primers with BioBrick prefix, T7-promoter and RBS in the fwd primer. Freiburg suffix in the rev primer.
- fwd: 5'-acgtgaattcgcggccgcttctagagtaatacgactcactatagggaaagaggagaaaaatggaatcaatcgaacaac-3'
- rev: 5'-acgtctgcagcggccgctactagtattaaccggtgctacccagcaaacgc-3'