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pQE-TriSystem His-Strep1 vector (V011598)

Price Information

Cat No. Plasmid Name Availability Add to cart
V011598 pQE-TriSystem His-Strep1 In stock, 1 week for quality controls

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Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.

Basic Vector Information

Vector Name:
pQE-TriSystem His-Strep1
Antibiotic Resistance:
Ampicillin
Length:
5760 bp
Type:
Insect Cell Vectors
Replication origin:
ori
Source/Author:
Qiagen
Copy Number:
High copy number
Promoter:
chicken β-actin

pQE-TriSystem His-Strep1 vector Map

Copy Sequence View Map
pQE-TriSystem His-Strep15760 bp60012001800240030003600420048005400CMV enhancerchicken beta-actin promoterExon 1intronT5 promoterp10 promoterRBSMCSStrep-Tag II8xHisstop codonsbeta-globin poly(A) signalT7 terminatorcontains part of ORF1629oriAmpRcontains ORF603 and part of lef2

Plasmid 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 ℃.

6. The concentration of plasmid re-measurement sometimes differs from the nominal value, which may be due to the position of the lyophilized plasmid in the tube, the efficiency of the re-dissolution, the measurement bias, and adsorption on the wall of the tube, therefore, it is recommended to transform and extract the plasmid before using it

General Plasmid Transform Protocol

1. Take one 100μl of the competent cells and thaw it on ice for 10min, add 2μl of plasmid, then ice bath for 30min, then heat-shock it at 42℃ for 60s, do not stir, and then ice bath for 2min.

2. Add 900μl of LB liquid medium without antibiotics, and incubate at 37℃ for 45min (30℃ for 1-1.5 hours) with 180rpm shaking.

3. Centrifuge at 6000rpm for 5min, leave only 100μl of supernatant to resuspend the bacterial precipitate and spread it onto the target plasmid-resistant LB plate.

4. Invert the plate and incubate at 37℃ for 14h, or at 30℃ for 20h.

5. Pick a single colony into LB liquid medium, add the corresponding antibiotics, incubate at 220rpm for 14h, and extract the plasmid according to the experimental needs and the instructions of the plasmid extraction kit.

pQE-TriSystem His-Strep1 vector Sequence

LOCUS       pQE-TriSystem_Hi        5760 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Vector for expression in bacterial, insect, and vertebrate cells of 
            proteins with C-terminal Strep-Tag II and 8xHis tags.
ACCESSION   .
VERSION     .
KEYWORDS    pQE-TriSystem His-Strep1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5760)
  AUTHORS   Qiagen
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 5760)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5760
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        64..367
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        373..650
                     /label=chicken beta-actin promoter
     exon            651..742
                     /note="Exon 1"
     intron          743..1004
     promoter        1073..1116
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA
                     polymerase, with embedded lac operator"
     promoter        1143..1252
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     RBS             1255..1260
                     /note="ribosome binding site"
     misc_feature    1265..1372
                     /label=MCS
                     /note="MCS"
                     /note="multiple cloning site"
     CDS             1267..1269
                     /codon_start=1
                     /product="start codon"
                     /label=start codon
                     /note="ATG"
                     /translation="M"
     CDS             1381..1404
                     /label=Strep-Tag II
                     /note="peptide that binds Strep-Tactin(R), an engineered 
                     form
                     of streptavidin"
     CDS             1411..1434
                     /label=8xHis
                     /note="8xHis affinity tag"
     misc_feature    1435..1445
                     /label=stop codons
                     /note="stop codons"
                     /note="stop codons in all three reading frames"
     polyA_signal    1589..1644
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     terminator      1732..1779
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_recomb     1792..3250
                     /label=baculovirus recombination region (ORF1629)
                     /note="contains part of ORF1629"
     rep_origin      complement(3409..3996)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4012..4869)
                     /label=AmpR
                     /note="beta-lactamase"
     misc_recomb     4935..5760
                     /label=baculovirus recombination region (lef2/ORF603)
                     /note="contains ORF603 and part of lef2"