Basic Vector Information
- Vector Name:
- pHUE
- Antibiotic Resistance:
- Ampicillin
- Length:
- 5942 bp
- Type:
- Cloning Vectors
- Replication origin:
- ori
- Source/Author:
- Catanzariti,A.M., Soboleva,T.A., Jans,D.A., Board,P.G. and Baker,R.T.
- Copy Number:
- High copy number
- Promoter:
- T7
- Cloning Method:
- Enzyme Cut
- 5' Primer:
- T7 promoter
- 3' Primer:
- T7 terminator
- Fusion Tag:
- UBA52
- Expression Method:
- IPTG induced
pHUE 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 ℃.
pHUE vector Sequence
LOCUS pHUE. 5942 bp DNA circular SYN 28-NOV-2018
DEFINITION Bacterial vector for inducible expression of N-terminally
6xHis-tagged proteins.
ACCESSION .
VERSION .
KEYWORDS pHUE.
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5942)
AUTHORS Novagen (EMD Millipore)
TITLE Direct Submission
REFERENCE 2 (bases 1 to 5942)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..5942
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 156..174
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 175..199
/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)."
RBS 214..236
/label=RBS
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 243..245
/codon_start=1
/product="start codon"
/label=start codon
/note="ATG"
/translation="M"
CDS 255..272
/label=6xHis
/note="6xHis affinity tag"
CDS 282..299
/label=thrombin site
/note="thrombin recognition and cleavage site"
CDS 303..530
/label=ubiquitin
/note="human ubiquitin C"
misc_feature 528..577
/label=MCS
/note="MCS"
/note="multiple cloning site"
terminator 636..683
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
promoter 972..1076
/label=AmpR promoter
CDS 1077..1934
/label=AmpR
/note="beta-lactamase"
rep_origin 2108..2696
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
misc_feature complement(2882..3024)
/label=bom
/note="basis of mobility region from pBR322"
CDS complement(3129..3317)
/label=rop
/note="Rop protein, which maintains plasmids at low copy
number"
protein_bind complement(4595..4616)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
CDS complement(4632..5711)
/label=lacI
/note="lac repressor"
promoter complement(5712..5789)
/label=lacI promoter
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