pET-19b vector (V011026)

Basic Vector Information

Vector Name:
pET-19b
Antibiotic Resistance:
Ampicillin
Length:
5717 bp
Type:
pET & Duet Vectors (Novagen)
Replication origin:
ori
Source/Author:
Novagen (EMD Millipore)
Copy Number:
High copy number

pET-19b vector Vector Map

pET-19b5717 bp60012001800240030003600420048005400tet promoterT7 terminatorMCSenterokinase site9xHisATGRBSlac operatorT7 promoterlacI promoterlacICAP binding siteropbomoriAmpRAmpR promoter

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 ℃.

pET-19b vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       pET-19b.        5717 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Bacterial vector for inducible expression of N-terminally 
            10xHis-tagged proteins with an enterokinase site.
ACCESSION   .
VERSION     .
KEYWORDS    pET-19b.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5717)
  AUTHORS   Novagen (EMD Millipore)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 5717)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5717
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        10..38
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
     terminator      complement(213..260)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    319..335
                     /label=MCS
                     /note="MCS"
                     /note="multiple cloning site"
     CDS             complement(336..350)
                     /label=enterokinase site
                     /note="enterokinase recognition and cleavage site"
     CDS             complement(369..395)
                     /label=9xHis
                     /note="9xHis affinity tag"
     CDS             complement(399..401)
                     /codon_start=1
                     /product="start codon"
                     /label=start codon
                     /note="ATG"
                     /translation="M"
     RBS             complement(408..430)
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    complement(445..469)
                     /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)."
     promoter        complement(470..488)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        797..874
                     /label=lacI promoter
     CDS             875..1954
                     /label=lacI
                     /note="lac repressor"
     protein_bind    1970..1991
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             3269..3457
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     misc_feature    3562..3704
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(3890..4478)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4652..5509)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(5510..5614)
                     /label=AmpR promoter

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