Basic Vector Information
- Vector Name:
- pN-PURO-PTP
- Antibiotic Resistance:
- Ampicillin
- Length:
- 5650 bp
- Type:
- Transfection vector
- Replication origin:
- ori
- Source/Author:
- Schimanski B, Nguyen TN, Gunzl A.
pN-PURO-PTP 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 ℃.
pN-PURO-PTP vector Sequence
LOCUS 40924_32963 5650 bp DNA circular SYN 18-DEC-2018 DEFINITION Transfection vector pN-PURO-PTP, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5650) AUTHORS Schimanski B, Nguyen TN, Gunzl A. TITLE Highly efficient tandem affinity purification of trypanosome protein complexes based on a novel epitope combination JOURNAL Eukaryotic Cell 4 (11), 1942-1950 (2005) PUBMED 16278461 REFERENCE 2 (bases 1 to 5650) AUTHORS Schimanski B, Nguyen TN, Gunzl A. TITLE Direct Submission JOURNAL Submitted (17-AUG-2005) Department of Genetics and Developmental Biology, University of Connecticut Health Center, 263 Farmington Ave, Farmington, CT 06030, USA REFERENCE 3 (bases 1 to 5650) TITLE Direct Submission REFERENCE 4 (bases 1 to 5650) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Eukaryotic Cell"; date: "2005"; volume: "4"; issue: "11"; pages: "1942-1950" COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted (17-AUG-2005) Department of Genetics and Developmental Biology, University of Connecticut Health Center, 263 Farmington Ave, Farmington, CT 06030, USA" COMMENT SGRef: number: 3; type: "Journal Article" FEATURES Location/Qualifiers source 1..5650 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 21..125 /label=AmpR promoter CDS 126..983 /label=AmpR /note="beta-lactamase" rep_origin 1157..1745 /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" protein_bind 2033..2054 /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." promoter 2069..2099 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2107..2123 /label=lac operator /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." primer_bind 2131..2147 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" promoter 2168..2186 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" misc_feature 2199..2204 /note="unique SacI restriction site for cloning the resistance marker cassette" misc_feature 2204..2702 /label=HSP70 genes 2 and 3 intergenic region /note="HSP70 genes 2 and 3 intergenic region" CDS 2703..3299 /label=PuroR /note="puromycin N-acetyltransferase" misc_feature 3315..4055 /label=beta-alpha tubulin intergenic region /note="beta-alpha tubulin intergenic region" misc_feature 4056..4061 /note="unique HindIII restriction site for cloning the resistance marker cassette and the PTP cassette" CDS 4526..4699 /label=ProtA /note="IgG-binding unit of Staphylococcus aureus protein A" CDS 4703..4873 /codon_start=1 /product="IgG-binding unit of Staphylococcus aureus protein A" /label=ProtA /translation="DNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLLA EAKKLNDAQAPK" CDS 4910..4930 /label=TEV site /note="tobacco etch virus (TEV) protease recognition and cleavage site" misc_feature 4973..4998 /note="multiple cloning site containing unique restriction sites for KpnI, ApaI, SnaBI and NotI for fusing the target protein to the PTP sequence" promoter complement(5007..5025) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(5032..5048) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" rep_origin 5190..5645 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis"
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