04/14/13
- New vector: design new vector for mammalian expression
- Order oligos
New mammalian expression vector MV9 and MV10
- Use the Oligator tool to design oligo-assembly inserts for "MV6" (pcDNA3.1+ puro, mammalian transfection vector; CMV promoter)
- MV9 - will carry Kozak-XbaI-NLS-6His-stop (for Brady's and Behzad's projects)
- Oligo assembly method will be used to make double stranded DNA with SpeI overhangs
- Insert will be ligated to XbaI-cut MV6 to destroy the preexisting XbaI site
- Clones will be screened for proper orientation and insert copy number
Oligator results:
- Custom prefix top = ctagt
- Custom prefix bottom = a
- Custom suffix top = a
- Custom suffix bottom = tgatc
CTAGTCCCGCCGCCACCATGGAGTCTAGACCCAAGAAAAAGCGCAAGGTACACCATCACCACCATCACGCGTAAAGCTGAGA
AGGGCGGCGGTGGTACCTCAGATCTGGGTTCTTTTTCGCGTTCCATGTGGTAGTGGTGGTAGTGCGCATTTCGACTCTGATC
- 30-mer 5'-ctagtCCCGCCGCCACCATGGAGTCTAGAC
- 52-mer 5'-CCAAGAAAAAGCGCAAGGTACACCATCACCACCATCACGCGTAAAGCTGAGa
- 52-mer 5'-ATGGTGTACCTTGCGCTTTTTCTTGGGTCTAGACTCCATGGTGGCGGCGGGA
- 30-mer 5'-CTAGTCTCAGCTTTACGCGTGATGGTGGTG
|