pHUE vector (V010154)

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 Map

pHUE5942 bp60012001800240030003600420048005400T7 promoterlac operatorRBSATG6xHisthrombin siteubiquitinT7 terminatorAmpR promoterAmpRoribomropCAP binding sitelacIlacI promoter

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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