Basic Vector Information
- Vector Name:
- pPIC3.5K-AOX1-EGFP
- Antibiotic Resistance:
- Ampicillin
- Length:
- 9919 bp
- Type:
- Protein expression
- Replication origin:
- ori
- Host:
- Yeast
- Selection Marker:
- HIS4
- Promoter:
- AOX1
- Growth Strain(s):
- DH5a
- Growth Temperature:
- 37℃
pPIC3.5K-AOX1-EGFP vector Vector Map
Plasmid Resuspension Protocol:
1. Centrifuge at 5,000×g for 5 min.
2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.
3. Close the tube and incubate for 10 minutes at room temperature.
4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.
5.Store the plasmid at -20 ℃.
pPIC3.5K-AOX1-EGFP vector Sequence
LOCUS 62056_18460 9919 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9919) AUTHORS . TITLE Direct Submission FEATURES Location/Qualifiers source 1..9919 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 2..937 /label=AOX1 promoter /note="inducible promoter, regulated by methanol" CDS 944..1660 /codon_start=1 /label=EGFP /note="enhanced GFP" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" terminator 1751..1997 /label=AOX1 terminator /note="transcription terminator for AOX1" CDS complement(5361..6380) /codon_start=1 /label=HygR /note="aminoglycoside phosphotransferase from E. coli" /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE " misc_feature 6765..7521 /label=AOX1 3' fragment /note="region downstream of Pichia pastoris AOX1 gene" misc_feature 7664..7804 /label=bom /note="basis of mobility region from pBR322" rep_origin complement(7990..8578) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(8752..9609) /codon_start=1 /label=AmpR /note="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(9610..9714) /label=AmpR promoter
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