SIRT6 Flag vector (V007170)

Basic Vector Information

      • Vector Name:
      • SIRT6 Flag
      • Antibiotic Resistance:
      • Ampicillin
      • Length:
      • 6486 bp
      • Type:
      • Mammalian Expression
      • Selection Marker:
      • Neomycin (select with G418)
      • Copy Number:
      • High Copy
      • Cloning Method:
      • Restriction Enzyme
      • 5' Primer:
      • T7
      • 3' Primer:
      • BGH Reverse

SIRT6 Flag vector Vector Map

SIRT6 Flag6486 bp300600900120015001800210024002700300033003600390042004500480051005400570060006300FLAGT7CMV promoterCMV enhancerpRS-markerAmpR promoterAmpRoriL4440CAP binding sitelac promoterlac operatorM13 revSV40 poly(A) signalNeoR/KanRSV40 promoterf1 oribGH poly(A) signal

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 ℃.

SIRT6 Flag vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported                6486 bp ds-DNA     circular SYN 20-DEC-2021
DEFINITION  Mammalian expression of human SIRT6 with flag tag.
ACCESSION   .
VERSION     .
KEYWORDS    SIRT6 Flag
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6486)
  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 6486)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol Cell. 
            2003 Feb . 11(2):437-44."
FEATURES             Location/Qualifiers
     source          1..6486
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             complement(9..32)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(1169..1188)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1170..1188)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(1233..1436)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1262..1282)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1437..1816
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     1988..2007
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        2082..2186
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2187..3047
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2405..2424)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3218..3806
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3707..3726
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3960..3977
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4094..4115
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4130..4160
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4168..4184
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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)."
     primer_bind     4173..4195
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4192..4208
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4192..4208
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    4257..4378
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4268..4287)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4322..4341
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(4552..5346)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4663..4682)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5273..5292
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(5413..5742)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5427..5562
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5481..5500)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     5727..5747
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(5756..6184)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5866..5887)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6078..6097
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    6230..6454
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     6443..6460
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"

This page is informational only.