Basic Vector Information
p3xFLAG-CMV-14 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 ℃.
p3xFLAG-CMV-14 vector Sequence
LOCUS p3xFLAG-CMV-14. 6310 bp DNA circular SYN 01-JAN-1980 DEFINITION Vector with a neomycin resistance marker for expression of C-terminally 3xFLAG-tagged proteins in mammalian cells. ACCESSION . VERSION . KEYWORDS p3xFLAG-CMV-14. SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6310) AUTHORS Sigma-Aldrich TITLE Direct Submission REFERENCE 2 (bases 1 to 6310) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article" FEATURES Location/Qualifiers source 1..6310 /mol_type="other DNA" /organism="synthetic DNA construct" primer_bind 141..157 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" enhancer 318..697 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 698..901 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" misc_feature 924..990 /label=MCS /note="MCS" /note="multiple cloning site" CDS 993..1058 /label=3xFLAG /note="three tandem FLAG(R) epitope tags, followed by an enterokinase cleavage site" polyA_signal 1069..1691 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" promoter 1733..2030 /label=SV40 promoter /note="SV40 enhancer and early promoter" CDS 2084..2875 /label=NeoR/KanR /note="aminoglycoside phosphotransferase" polyA_signal 3530..3664 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" promoter complement(3670..3688) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(3702..3718) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind complement(3726..3742) /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 complement(3750..3780) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind complement(3795..3816) /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(4104..4692) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(4866..5723) /label=AmpR /note="beta-lactamase" promoter complement(5724..5828) /label=AmpR promoter rep_origin 5855..6310 /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis"
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