Basic Vector Information
pTriEx-2 Neo vector Vector Map
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 Neo vector Sequence
LOCUS pTriEx-2_Neo. 6820 bp DNA circular SYN 01-JAN-1980
DEFINITION Insect, bacterial, and mammalian vector with a neomycin resistance
marker, encoding an N-terminal 6xHis-S-Tag cassette and a C-terminal
HSV-8xHis cassette.
ACCESSION .
VERSION .
KEYWORDS pTriEx-2 Neo.
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6820)
AUTHORS Novagen (EMD Millipore)
TITLE Direct Submission
REFERENCE 2 (bases 1 to 6820)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..6820
/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"
misc_feature 2761..3223
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
CDS 3259..4059
/label=NeoR/KanR
/note="aminoglycoside phosphotransferase from Tn5"
polyA_signal 4180..4235
/label=beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal (Gil and
Proudfoot, 1987)"
terminator 4323..4370
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
misc_recomb 4383..5088
/label=baculovirus recombination region (ORF1629)
/note="contains part of ORF1629"
protein_bind complement(5097..5130)
/label=loxP
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
rep_origin complement(5298..5885)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5901..6758)
/label=AmpR
/note="beta-lactamase"
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