Flag-SIRT1 H363Y vector (V000710)

Basic Vector Information

      • Vector Name:
      • Flag-SIRT1 H363Y
      • Antibiotic Resistance:
      • Ampicillin
      • Length:
      • 5420 bp
      • Type:
      • Mammalian Expression
      • Copy Number:
      • High Copy
      • Cloning Method:
      • Restriction Enzyme
      • 5' Primer:
      • SV40pro-F
      • 3' Primer:
      • SV40pA-R

Flag-SIRT1 H363Y vector Vector Map

Flag-SIRT1 H363Y5420 bp60012001800240030003600420048005400FLAGSV40pro-FpBABE 3'AmpR promoterAmpRoriL4440pBRrevBamSV40 poly(A) signalIn lacZ geneT7SIRT1

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 H363Y vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported                5420 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    Flag-SIRT1 H363Y
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"
     CDS             complement(1420..1443)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(1534..1553)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     1772..1792
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1847..1951
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1952..2812
                     /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     complement(2170..2189)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2983..3571
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3472..3491
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3725..3742
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     4007..4026
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     polyA_signal    4085..4219
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4109..4128)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4163..4182
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4455..4477
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4469..4486
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4470..4486
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4493..4512
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4493..4511
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(join(4587..5420,1..1407))
                     /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
                     NYTQNIDTLEQVAGIQRIIQCYGSFATASCLICKYKVDCEAVRGDIFNQVVPRCPRCPA
                     DEPLAIMKPEIVFFGENLPEQFHRAMKYDKDEVDLLIVIGSSLKVRPVALIPSSIPHEV
                     PQILINREPLPHLHFDVELLGDCDVIINELCHRLGGEYAKLCCNPVKLSEITEKPPRTQ
                     KELAYLSELPPTPLHVSEDSSSPERTSPPDSSVIVTLLDQAAKSNDDLDVSESKGCMEE
                     KPQEVQTSRNVESIAEQMENPDLKNVGSSTGEKNERTSVAGTVRKCWPNRVAKEQISRR
                     LDGNQYLFLPPNRYIFHGAEVYSDSEDDVLSSSSCGSNSDSGTCQSPSLEEPMEDESEI
                     EEFYNGLEDEPDVPERAGGAGFGTDGDDQEAINEAISVKQEVTDMNYPSNKS"

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