Basic Vector Information
- Vector Name:
- SIRT5 Flag
- Antibiotic Resistance:
- Ampicillin
- Length:
- 6351 bp
- Type:
- Mammalian Expression
- Replication origin:
- ori
- Selection Marker:
- Neomycin (select with G418)
- Copy Number:
- High Copy
- Cloning Method:
- Restriction Enzyme
- 5' Primer:
- T7
- 3' Primer:
- BGH Reverse
SIRT5 Flag 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 ℃.
SIRT5 Flag vector Sequence
LOCUS SIRT5_Flag. 6351 bp DNA circular SYN 13-MAY-2021
DEFINITION Mammalian expression of human SIRT5 with flag tag.
ACCESSION .
VERSION .
KEYWORDS SIRT5 Flag.
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6351)
AUTHORS North BJ, Marshall BL, Borra MT, Denu JM, Verdin E
TITLE The human Sir2 ortholog, SIRT2, is an NAD+-dependent tubulin
deacetylase.
JOURNAL Mol Cell. 2003 Feb . 11(2):437-44.
PUBMED 12620231
REFERENCE 2 (bases 1 to 6351)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 6351)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Mol Cell.
2003 Feb . 11(2):437-44."
COMMENT SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..6351
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind complement(63..80)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(234..822)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(996..1853)
/label=AmpR
/note="beta-lactamase"
promoter complement(1854..1958)
/label=AmpR promoter
primer_bind complement(2033..2052)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 2224..2603
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 2604..2807
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
promoter 2852..2870
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 2931..3860
/gene="SIRT5"
/label=SIRT5
/note="NAD-dependent protein deacylase sirtuin-5,
mitochondrial from Homo sapiens. Accession#: Q9NXA8"
CDS 3873..3896
/label=FLAG
/note="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
polyA_signal 3937..4161
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 4207..4635
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 4649..4978
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
CDS 5045..5836
/label=NeoR/KanR
/note="aminoglycoside phosphotransferase"
polyA_signal 6013..6146
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(6183..6199)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(6207..6223)
/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(6231..6261)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(6276..6297)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
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