Flag-SIRT1 vector (V006858)

Basic Vector Information

      • Vector Name:
      • Flag-SIRT1
      • Antibiotic Resistance:
      • Ampicillin
      • Length:
      • 5420 bp
      • Type:
      • Mammalian Expression
      • Copy Number:
      • High Copy
      • Cloning Method:
      • Restriction Enzyme
      • 5' Primer:
      • SV40pro-F

Flag-SIRT1 vector Vector Map

Flag-SIRT15420 bp60012001800240030003600420048005400pBABE 3'SV40pro-FFLAGSIRT1T7M13 ForwardSV40 poly(A) signalpBRrevBamL4440oriAmpRAmpR promoter

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

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported                5420 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Mammalian expression of flag-tagged SIRT1.
ACCESSION   .
VERSION     .
KEYWORDS    Flag-SIRT1
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)
  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"
FEATURES             Location/Qualifiers
     source          1..5420
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(55..75)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     primer_bind     294..313
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             404..427
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             440..2680
                     /codon_start=1
                     /product="human NAD+-dependent protein deacetylase 
                     sirtuin-1"
                     /label=SIRT1
                     /note="homolog of budding yeast Sir2"
                     /translation="ADEAALALQPGGSPSAAGADREAASSPAGEPLRKRPRRDGPGLER
                     SPGEPGGAAPEREVPAAARGCPGAAAAALWREAEAEAAAAGGEQEAQATAAAGEGDNGP
                     GLQGPSREPPLADNLYDEDDDDEGEEEEEAAAAAIGYRDNLLFGDEIITNGFHSCESDE
                     EDRASHASSSDWTPRPRIGPYTFVQQHLMIGTDPRTILKDLLPETIPPPELDDMTLWQI
                     VINILSEPPKRKKRKDINTIEDAVKLLQECKKIIVLTGAGVSVSCGIPDFRSRDGIYAR
                     LAVDFPDLPDPQAMFDIEYFRKDPRPFFKFAKEIYPGQFQPSLCHKFIALSDKEGKLLR
                     NYTQNIDTLEQVAGIQRIIQCHGSFATASCLICKYKVDCEAVRGDIFNQVVPRCPRCPA
                     DEPLAIMKPEIVFFGENLPEQFHRAMKYDKDEVDLLIVIGSSLKVRPVALIPSSIPHEV
                     PQILINREPLPHLHFDVELLGDCDVIINELCHRLGGEYAKLCCNPVKLSEITEKPPRTQ
                     KELAYLSELPPTPLHVSEDSSSPERTSPPDSSVIVTLLDQAAKSNDDLDVSESKGCMEE
                     KPQEVQTSRNVESIAEQMENPDLKNVGSSTGEKNERTSVAGTVRKCWPNRVAKEQISRR
                     LDGNQYLFLPPNRYIFHGAEVYSDSEDDVLSSSSCGSNSDSGTCQSPSLEEPMEDESEI
                     EEFYNGLEDEPDVPERAGGAGFGTDGDDQEAINEAISVKQEVTDMNYPSNKS"
     primer_bind     complement(2755..2774)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2756..2774)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2781..2798)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2781..2797)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2790..2812)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     polyA_signal    3048..3182
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3085..3104)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3139..3158
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     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"
     primer_bind     complement(3776..3795)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4455..5315)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5078..5097
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5316..5420)
                     /gene="bla"
                     /label=AmpR promoter

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