Difference between revisions of "Part:BBa P1014:Design"
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===Source=== | ===Source=== | ||
*Novel gentamicin resistance gene | *Novel gentamicin resistance gene | ||
| − | *Acquired by PCR from pTEX5500ts, | + | *Acquired by PCR from pTEX5500ts, Genbank DQ208936, kind gift of Barbara E. Murray. |
*pTEX5500ts was, in turn was derived from Genbank AF016483. | *pTEX5500ts was, in turn was derived from Genbank AF016483. | ||
Revision as of 17:20, 19 June 2006
gentamicin resistance cassette aph(2)
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
- PCR primers
- gtttcttcgaattcgcggccgcttctagag gagataggttatgcaagat,P1014-F
- gtttcttcctgcagcggccgctactagta gattccggattctaaaaaaagg,P1014-R2 (after terminator)
- gtttcttcctgcagcggccgctactagta ctatttaattttaatgcttctg,P1014-R3 (after stop codon)
- XbaI site at 660 removed by mutating c->t which also improves codon usage.
- gatgatttttacagggatattttagagaacgaaatctac,P1014-XF
- gtagatttcgttctctaaaatatccctgtaaaaatcatc,P1014-XR
Source
- Novel gentamicin resistance gene
- Acquired by PCR from pTEX5500ts, Genbank DQ208936, kind gift of Barbara E. Murray.
- pTEX5500ts was, in turn was derived from Genbank AF016483.
References
- Nallapareddy SR, Singh KV, Murray BE. Construction of improved temperature-sensitive and mobilizable vectors and their use for constructing mutations in the adhesin-encoding acm gene of poorly transformable clinical Enterococcus. Appl Environ Microbiol. 2006 Jan;72(1):334-45 faecium strains. PMID 16391062
- Tsai SF, Zervos MJ, Clewell DB, Donabedian SM, Sahm DF, Chow JW. A new high-level gentamicin resistance gene, aph(2)-Id, in Enterococcus spp. Antimicrob Agents Chemother. 1998 May;42(5):1229-32. PMID 9593155