pTriEx-2 vector (V011586)

Basic Vector Information

Vector Name:
pTriEx-2
Antibiotic Resistance:
Ampicillin
Length:
5457 bp
Type:
Insect Cell Vectors
Replication origin:
ori
Source/Author:
Novagen (EMD Millipore)
Copy Number:
High copy number
Promoter:
chicken β-actin

pTriEx-2 vector Vector Map

pTriEx-25457 bp60012001800240030003600420048005400contains ORF603 and part of lef2CMV enhancerchicken beta-actin promoterExon 1T7 promoterlac operatorp10 promoterRBSATG6xHisS-Tagthrombin siteenterokinase siteHSV tag8xHisbeta-globin poly(A) signalT7 terminatorcontains part of ORF1629loxPoriAmpR

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 ℃.

pTriEx-2 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       pTriEx-2.        5457 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Insect, bacterial, and mammalian vector for expressing proteins with
            a cleavable N-terminal 6xHis-S-Tag cassette and a C-terminal 
            HSV-8xHis cassette.
ACCESSION   .
VERSION     .
KEYWORDS    pTriEx-2.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5457)
  AUTHORS   Novagen (EMD Millipore)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 5457)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5457
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_recomb     181..1008
                     /label=baculovirus recombination region (lef2/ORF603)
                     /note="contains ORF603 and part of lef2"
     enhancer        1140..1443
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1449..1726
                     /label=chicken beta-actin promoter
     exon            1727..1819
                     /note="Exon 1"
     intron          1820..2081
     promoter        2150..2168
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2169..2193
                     /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        2209..2318
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     RBS             2320..2325
                     /note="ribosome binding site"
     CDS             2333..2335
                     /codon_start=1
                     /product="start codon"
                     /label=start codon
                     /note="ATG"
                     /translation="M"
     CDS             2339..2356
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             2366..2410
                     /label=S-Tag
                     /note="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
     CDS             2426..2443
                     /label=thrombin site
                     /note="thrombin recognition and cleavage site"
     CDS             2462..2476
                     /label=enterokinase site
                     /note="enterokinase recognition and cleavage site"
     CDS             2600..2632
                     /label=HSV tag
                     /note="HSV (herpes simplex virus) epitope tag"
     CDS             2639..2662
                     /label=8xHis
                     /note="8xHis affinity tag"
     polyA_signal    2817..2872
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     terminator      2960..3007
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_recomb     3020..3725
                     /label=baculovirus recombination region (ORF1629)
                     /note="contains part of ORF1629"
     protein_bind    complement(3734..3767)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     rep_origin      complement(3935..4522)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4538..5395)
                     /label=AmpR
                     /note="beta-lactamase"

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