Basic Vector Information
TAGZyme pQE-1 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-1 vector Sequence
LOCUS TAGZyme_pQE-1. 3466 bp DNA circular SYN 01-JAN-1980
DEFINITION Vector for bacterial expression of 6xHis-tagged proteins that can be
processed by TAGZyme.
ACCESSION .
VERSION .
KEYWORDS TAGZyme pQE-1.
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3466)
AUTHORS Qiagen
TITLE Direct Submission
REFERENCE 2 (bases 1 to 3466)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
COMMENT A blunt coding sequence can be inserted at the PvuII site, allowing
TAGZyme to remove the 6xHis tag and subsequent glutamine.
FEATURES Location/Qualifiers
source 1..3466
/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..138
/label=6xHis
/note="6xHis affinity tag"
misc_feature 139..197
/label=MCS
/note="MCS"
/note="multiple cloning site"
misc_feature 197..207
/label=stop codons
/note="stop codons"
/note="stop codons in all three reading frames"
terminator 213..307
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
CDS 351..1007
/label=CmR
/note="chloramphenicol acetyltransferase"
terminator 1075..1161
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
misc_feature 1316..1456
/label=bom
/note="basis of mobility region from pBR322"
rep_origin complement(1642..2230)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(2404..3261)
/label=AmpR
/note="beta-lactamase"
promoter complement(3262..3366)
/label=AmpR promoter
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