04/14/13
- New vector: design new vector for mammalian expression
- Order oligos
New mammalian expression vector MV9
- 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)
- The XbaI site will be used to accept X/S inserts; plasmid digests will be needed to check proper orientation
- 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
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