Basic Vector Information
pTriEx-4 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-4 vector Sequence
LOCUS pTriEx-4. 5238 bp DNA circular SYN 01-JAN-1980 DEFINITION Insect, bacterial, and mammalian vector with a CMV promoter, encoding an N-terminal 6xHis-S-Tag and a C-terminal HSV-8xHis. ACCESSION . VERSION . KEYWORDS pTriEx-4. SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5238) AUTHORS Novagen (EMD Millipore) TITLE Direct Submission REFERENCE 2 (bases 1 to 5238) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article" FEATURES Location/Qualifiers source 1..5238 /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 1076..1379 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 1381..1580 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" exon 1597..1698 /note="Exon 1" intron 1699..1862 promoter 1931..1949 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 1950..1974 /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 1990..2099 /label=p10 promoter /note="baculovirus promoter for expression in insect cells" RBS 2101..2106 /note="ribosome binding site" CDS 2114..2116 /codon_start=1 /product="start codon" /label=start codon /note="ATG" /translation="M" CDS 2120..2137 /label=6xHis /note="6xHis affinity tag" CDS 2147..2191 /label=S-Tag /note="affinity and epitope tag derived from pancreatic ribonuclease A" CDS 2207..2224 /label=thrombin site /note="thrombin recognition and cleavage site" CDS 2243..2257 /label=enterokinase site /note="enterokinase recognition and cleavage site" CDS 2381..2413 /label=HSV tag /note="HSV (herpes simplex virus) epitope tag" CDS 2420..2443 /label=8xHis /note="8xHis affinity tag" polyA_signal 2598..2653 /label=beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal (Gil and Proudfoot, 1987)" terminator 2741..2788 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" misc_recomb 2801..3506 /label=baculovirus recombination region (ORF1629) /note="contains part of ORF1629" protein_bind complement(3515..3548) /label=loxP /note="Cre-mediated recombination occurs in the 8-bp core sequence (ATGTATGC) (Shaw et al., 2021)." rep_origin complement(3716..4303) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(4319..5176) /label=AmpR /note="beta-lactamase"
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