Basic Vector Information
- Vector Name:
- Flag-SIRT1
- Antibiotic Resistance:
- Ampicillin
- Length:
- 5420 bp
- Type:
- Mammalian Expression
- Replication origin:
- ori
- Copy Number:
- High Copy
- Cloning Method:
- Restriction Enzyme
- 5' Primer:
- SV40pro-F
Flag-SIRT1 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 ℃.
Flag-SIRT1 vector Sequence
LOCUS 40924_835 5420 bp DNA circular SYN 13-MAY-2021
DEFINITION Mammalian expression of flag-tagged SIRT1.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5420)
AUTHORS Brunet A, Sweeney LB, Sturgill JF, Chua KF, Greer PL, Lin Y, Tran H,
Ross SE, Mostoslavsky R, Cohen HY, Hu LS, Cheng HL, Jedrychowski MP,
Gygi SP, Sinclair DA, Alt FW, Greenberg ME
TITLE Stress-dependent regulation of FOXO transcription factors by the
SIRT1 deacetylase.
JOURNAL Science 2004 Mar 26;303(5666):2011-5. Epub 2004 Feb 19.
PUBMED 14976264
REFERENCE 2 (bases 1 to 5420)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 5420)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Science";
date: "2004-03-26"; volume: "303(5666)"; pages: "2011-5. Epub 2004
Feb 19"
COMMENT SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..5420
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind complement(55..75)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 73..362
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
CDS 404..427
/codon_start=1
/label=FLAG
/note="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/translation="DYKDDDDK"
CDS 440..2677
/codon_start=1
/label=SIRT1
/note="human NAD+-dependent protein deacetylase sirtuin-1"
/translation="ADEAALALQPGGSPSAAGADREAASSPAGEPLRKRPRRDGPGLER
SPGEPGGAAPEREVPAAARGCPGAAAAALWREAEAEAAAAGGEQEAQATAAAGEGDNGP
GLQGPSREPPLADNLYDEDDDDEGEEEEEAAAAAIGYRDNLLFGDEIITNGFHSCESDE
EDRASHASSSDWTPRPRIGPYTFVQQHLMIGTDPRTILKDLLPETIPPPELDDMTLWQI
VINILSEPPKRKKRKDINTIEDAVKLLQECKKIIVLTGAGVSVSCGIPDFRSRDGIYAR
LAVDFPDLPDPQAMFDIEYFRKDPRPFFKFAKEIYPGQFQPSLCHKFIALSDKEGKLLR
NYTQNIDTLEQVAGIQRIIQCHGSFATASCLICKYKVDCEAVRGDIFNQVVPRCPRCPA
DEPLAIMKPEIVFFGENLPEQFHRAMKYDKDEVDLLIVIGSSLKVRPVALIPSSIPHEV
PQILINREPLPHLHFDVELLGDCDVIINELCHRLGGEYAKLCCNPVKLSEITEKPPRTQ
KELAYLSELPPTPLHVSEDSSSPERTSPPDSSVIVTLLDQAAKSNDDLDVSESKGCMEE
KPQEVQTSRNVESIAEQMENPDLKNVGSSTGEKNERTSVAGTVRKCWPNRVAKEQISRR
LDGNQYLFLPPNRYIFHGAEVYSDSEDDVLSSSSCGSNSDSGTCQSPSLEEPMEDESEI
EEFYNGLEDEPDVPERAGGAGFGTDGDDQEAINEAISVKQEVTDMNYPSNKS"
promoter complement(2756..2774)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(2781..2797)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
polyA_signal 3048..3182
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3241..3260)
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
primer_bind complement(3525..3542)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3696..4284)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4458..5315)
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(5316..5420)
/label=AmpR promoter
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