Basic Vector Information
- Vector Name:
- pET-3d
- Antibiotic Resistance:
- Ampicillin
- Length:
- 4637 bp
- Type:
- pET & Duet Vectors (Novagen)
- Replication origin:
- ori
- Source/Author:
- Novagen (EMD Millipore)
- Copy Number:
- High copy number
- Promoter:
- tet
pET-3d 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 ℃.
pET-3d vector Sequence
LOCUS 40924_17951 4637 bp DNA circular SYN 01-JAN-1980 DEFINITION Bacterial expression vector with a BamHI cloning site. Same reading frame as pET-3c but with an NcoI site at the start codon. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4637) AUTHORS Novagen (EMD Millipore) TITLE Direct Submission REFERENCE 2 (bases 1 to 4637) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article" FEATURES Location/Qualifiers source 1..4637 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 10..38 /label=tet promoter /note="E. coli promoter for tetracycline efflux protein gene" terminator complement(404..451) /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" CDS complement(517..549) /codon_start=1 /label=T7 tag (gene 10 leader) /note="leader peptide from bacteriophage T7 gene 10" /translation="MASMTGGQQMG" RBS complement(556..578) /label=RBS /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" promoter complement(610..628) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 2189..2377 /codon_start=1 /label=rop /note="Rop protein, which maintains plasmids at low copy number" /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" misc_feature 2482..2624 /label=bom /note="basis of mobility region from pBR322" rep_origin complement(2810..3398) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(3572..4429) /codon_start=1 /label=AmpR /note="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(4430..4534) /label=AmpR promoter
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