Basic Vector Information
- Vector Name:
- pMETα B
- Antibiotic Resistance:
- Ampicillin
- Length:
- 8049 bp
- Type:
- Yeast Expression Vectors
- Replication origin:
- ori
- Selection Marker:
- ADE2
- Copy Number:
- High copy number
- Promoter:
- ADE2
- Cloning Method:
- Enzyme digestion and ligation
- 5' Primer:
- AUG1-F: 5´-CAATTTACATCTTTATTTATTAACG-3´; alpha-Factor-F: 5´-TACTATTGCCAGCATTGCTGC-3´
- 3' Primer:
- AUG1-R: 5´-GAAGAGAAAAACATTAGTTGGC-3´
- Fusion Tag:
- C-V5, C-His
pMETα B 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 ℃.
pMETα B vector Sequence
LOCUS 40924_30650 8049 bp DNA circular SYN 13-JAN-2022 DEFINITION synthetic circular DNA. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8049) TITLE Direct Submission REFERENCE 2 (bases 1 to 8049) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article" FEATURES Location/Qualifiers source 1..8049 /mol_type="other DNA" /organism="synthetic DNA construct" CDS 1197..1463 /codon_start=1 /label=alpha-factor secretion signal /note="N-terminal secretion signal from S. cerevisiae alpha-factor" /translation="MRFPSIFTAVLFAASSALAAPVNTTTEDETAQIPAEAVIGYSDLE GDFDVAVLPFSNSTNNGLLFINTTIASIAAKEEGVSLEKREAEA" CDS 1538..1579 /codon_start=1 /label=V5 tag /note="epitope tag from simian virus 5" /translation="GKPIPNPLLGLDST" CDS 1589..1606 /codon_start=1 /label=6xHis /note="6xHis affinity tag" /translation="HHHHHH" promoter 2714..3089 /label=ADE2 promoter CDS 3090..4802 /codon_start=1 /label=ADE2 /note="phosphoribosylaminoimidazole carboxylase, required for adenine biosynthesis" /translation="MDSRTVGILGGGQLGRMIVEAANRLNIKTVILDAENSPAKQISNS NDHVNGSFSNPLDIEKLAEKCDVLTIEIEHVDVPTLKNLQVKHPKLKIYPSPETIRLIQ DKYIQKEHLIKNGIAVTQSVPVEQASETSLLNVGRDLGFPFVLKSRTLAYDGRGNFVVK NKEMIPEALEVLKDRPLYAEKWAPFTKELAVMIVRSVNGLVFSYPIVETIHKDNICDLC YAPARVPDSVQLKAKLLAENAIKSFPGCGIFGVEMFYLETGELLINEIAPRPHNSGHYT IDACVTSQFEAHLRSILDLPMPKNFTSFSTITTNAIMLNVLGDKHTKDKELETCERALA TPGSSVYLYGKESRPNRKVGHINIIASSMAECEQRLNYITGRTDIPIKISVAQKLDLEA MVKPLVGIIMGSDSDLPVMSAACAVLKDFGVPFEVTIVSAHRTPHRMSAYAISASKRGI KTIIAGAGGAAHLPGMVAAMTPLPVIGVPVKGSCLDGVDSLHSIVQMPRGVPVATVAIN NSTNAALLAVRLLGAYDSSYTTKMEQFLLKQEEEVLVKAQKLETVGYEAYLENK" rep_origin complement(6021..6609) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(6783..7640) /codon_start=1 /label=AmpR /note="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(7641..7745) /label=AmpR promoter
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