Basic Vector Information
TAGZyme pQE-2 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 ℃.
TAGZyme pQE-2 vector Sequence
LOCUS TAGZyme_pQE-2. 4758 bp DNA circular SYN 01-JAN-1980 DEFINITION Vector with a lacIq repressor gene for bacterial expression of 6xHis-tagged proteins that can be processed by TAGZyme. ACCESSION . VERSION . KEYWORDS TAGZyme pQE-2. SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4758) AUTHORS Qiagen TITLE Direct Submission REFERENCE 2 (bases 1 to 4758) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article" COMMENT For TAGZyme cleavage, a glutamine should be inserted at an uneven amino acid position, followed by the first amino acid of the expressed protein. FEATURES Location/Qualifiers source 1..4758 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 10..54 /label=T5 promoter /note="bacteriophage T5 promoter for E. coli RNA polymerase, with embedded lac operator" protein_bind 62..78 /label=lac repressor encoded by lacI binding site /bound_moiety="lac repressor encoded by lacI" /note="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)." RBS 101..106 /note="ribosome binding site" CDS 115..117 /codon_start=1 /product="start codon" /label=start codon /note="ATG" /translation="M" CDS 121..141 /label=7xHis /note="6xHis affinity tag" misc_feature 139..199 /label=MCS /note="MCS" /note="multiple cloning site" misc_feature 199..209 /label=stop codons /note="stop codons" /note="stop codons in all three reading frames" terminator 215..309 /label=lambda t0 terminator /note="transcription terminator from phage lambda" CDS 353..1009 /label=CmR /note="chloramphenicol acetyltransferase" terminator 1077..1163 /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" protein_bind complement(1222..1243) /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." CDS complement(1259..2338) /label=lacI /note="lac repressor" promoter complement(2339..2416) /label=lacIq promoter /note="In the lacIq allele, a single base change in the promoter boosts expression of the lacI gene about 10-fold." misc_feature 2608..2748 /label=bom /note="basis of mobility region from pBR322" rep_origin complement(2934..3522) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(3696..4553) /label=AmpR /note="beta-lactamase" promoter complement(4554..4658) /label=AmpR promoter
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