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pLac-6×His-T7 RNA polymerase vector (V014808)

Basic Vector Information

Vector Name:
pLac-6×His-T7 RNA polymerase
Antibiotic Resistance:
Ampicillin
Length:
5413 bp
Type:
Protein expression
Replication origin:
ori
Host:
E. coli
Promoter:
Lac
Growth Temperature:
37℃

pLac-6×His-T7 RNA polymerase vector Vector Map

pLac-6×His-T7 RNA polymerase5413 bp60012001800240030003600420048005400CAP binding sitelac promoterlac operatorM13 revRBS6xHisenterokinase siteT7 RNA polymerase from Escherichia phage T7. Accession#: P00573tet promoterAmpR promoterAmpRori

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 ℃.

pLac-6×His-T7 RNA polymerase vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       62056_13650        5413 bp DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5413)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5413
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    288..309
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        324..354
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    362..378
                     /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)."
     primer_bind     386..402
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     RBS             451..473
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             492..509
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             519..533
                     /label=enterokinase site
                     /note="enterokinase recognition and cleavage site"
     CDS             534..3182
                     /note="T7 RNA polymerase from Escherichia phage T7.
                     Accession#: P00573"
     promoter        complement(3548..3576)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
     promoter        3689..3793
                     /label=AmpR promoter
     CDS             3794..4651
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      4825..5413
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"

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