Flag-SIRT1

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载体基本信息

质粒名称: Flag-SIRT1 
出品公司: 爱迪基因
目录编号: 1791
存储实验室: Michael Greenberg

载体骨架基本信息
载体名称: pECE
载体抗性: 氨苄霉素
空载体大小: 2877 bp
完整质粒大小: 5191 bp
载体类型: 哺乳细胞表达
高拷贝/低拷贝: 高拷贝
生长条件: 37 ℃

插入基因信息
插入基因名称: SIRT1
物种来源: H. sapiens (human)
插入大小: 2314 bp
基因又名: Sir2
GenBank编号 NM_012238 NP_036370
标签/融合蛋白: Flag (N terminal on insert)
克隆方法: 限制性内切酶
5' 克隆位点: HindIII(未破坏)
3' 克隆位点: XbaI (未破坏)
5' 测序引物 SV40pro-F

文献引用方法
材料和方法部分: Flag-SIRT1 was a gift from Michael Greenberg (爱迪基因 plasmid # 1791)
参考文献部分: Stress-dependent regulation of FOXO transcription factors by the SIRT1 deacetylase. 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. Science 2004 Mar 26;303(5666):2011-5. Epub 2004 Feb 19. 10.1126/science.1094637 PubMed 14976264

载体图谱质粒图谱和多克隆位点信息

Flag-SIRT1质粒图谱

载体简介

载体序列

LOCUS       Flag-SIRT1                5420 bp ds-DNA     circular SYN 10-JAN-2018
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
COMMENT     This file is created by Vector NTI
            http://www.biofeng.com/
COMMENT     ORIGDB|GenBank
COMMENT     VNTAUTHORNAME|biofeng.com|
COMMENT     VNTNAME|Flag-SIRT1|
FEATURES             Location/Qualifiers
     source          1..5420
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     255..274
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     polyA_signal    333..467
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(357..376)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     411..430
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     703..725
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     717..734
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     718..734
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     741..760
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        741..759
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(835..3075)
                     /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"
     CDS             complement(3088..3111)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(3202..3221)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     3440..3460
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3515..3619
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3620..4480
                     /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(3838..3857)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4651..5239
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5140..5159
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5393..5410
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatatcga
       61 cgctctccct tatgcgactc ctgcattagg aagcagccca gtagtaggtt gaggccgttg
      121 agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg cgcccaacag tcccccggcc
      181 acggggcctg ccaccatacc cacgccgaaa caagcgctca tgagcccgaa gtggcgagcc
      241 cgatcttccc catcggtgat gtcggcgata taggcgccag caaccgcacc tgtggcgccg
      301 gtgatgccgg ccacgatgcg tccggcgtag aggatccaga catgataaga tacattgatg
      361 agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg
      421 atgctattgc tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt
      481 gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa
      541 acctctacag atgtgatatg gctgattatg atcattactt atctaggtat aggctgcgca
      601 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg
      661 gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta
      721 aaacgacggc cagtgaattg taatacgact cactataggg cgaattgaat ttagcggccg
      781 cgaattcgcc cttgtctaga gtggaacaat tcctgtacct gcacaattat tacactatga
      841 tttgtttgat ggatagttca tgtctgttac ttcctgtttc acagatatag cttcattaat
      901 tgcctcttga tcatctccat cagtcccaaa tccagctcct ccagctctct ctggaacatc
      961 aggctcatct tctaagccat tgtagaattc ttcaatttca ctttcatcct ccatgggttc
     1021 ttctaaactt ggactctggc atgtcccact atcactgtta ctgccacaag aactagagga
     1081 taagacgtca tcttcagagt ctgaatatac ctcagcgcca tggaaaatgt aacgatttgg
     1141 tggcaaaaac agatactgat taccatcaag ccgcctacta atctgctcct ttgccactct
     1201 attaggccag cattttctca ctgttccagc cactgaagtt ctttcatttt tctccccagt
     1261 actagaacca acattcttca aatccggatt ttccatctgt tcagcaatac tttcaacatt
     1321 cctagaagtt tgtacttcct gtggtttttc ttccatacaa ccttttgatt cagacacatc
     1381 taaatcatca ttactcttag ctgcttggtc taaaagtgtg acaatcactg aagaatctgg
     1441 tggtgaagtt ctttctggtg aacttgagtc ttctgaaaca tgaagaggtg tgggtggcaa
     1501 ctctgacaaa taagccaatt ctttttgtgt tcgtggaggt ttttcagtaa tttctgaaag
     1561 ctttacaggg ttacagcaaa gtttggcata ttcaccacct aacctatgac acaattcatt
     1621 aattatgaca tcacagtctc caagaagctc tacatcaaaa tgcagatgag gcaaaggttc
     1681 tctattaatt aatatctgag gcacttcatg gggtatggaa cttggaatta gtgctactgg
     1741 tcttactttg agggaagacc caataacaat gaggaggtca acttcatctt tgtcatactt
     1801 catggctcta tgaaactgtt ctggtaaatt ttcaccaaaa aacacaatct ctggtttcat
     1861 gatagcaagc ggttcatcag ctgggcacct aggacatcga ggaactacct gattaaaaat
     1921 atctcctcgt acagcttcac agtcaacttt gtatttacaa atcaggcaag atgctgttgc
     1981 aaaggaacca tgacactgaa ttatcctttg gattcccgca acctgttcca gcgtgtctat
     2041 gttctgggta tagttgcgaa gtagttttcc ttccttatct gacaaggcta tgaatttgtg
     2101 acagagagat ggctggaatt gtccaggata tatttccttt gcaaacttga agaatggtct
     2161 tggatctttt ctgaaatatt caatatcaaa catcgcttga ggatctggaa gatctgggaa
     2221 gtctacagca aggcgagcat aaataccatc ccttgacctg aagtcaggta ttccacatga
     2281 aacagacacc ccagctccag ttagaactat aatttttttg cactcttgca gtaatttcac
     2341 agcatcttca attgtattaa tatcttttct ttttttcctt tttggtggtt ctgaaaggat
     2401 attaataaca atctgccaca gtgtcatatc atccaactca ggtggaggta ttgtttccgg
     2461 caataaatct ttaagaattg ttcgaggatc tgtgccaatc ataagatgtt gctgaacaaa
     2521 agtatatgga cctatccgtg gccttggagt ccagtcacta gagcttgcat gtgaggctct
     2581 atcctcctca tcactttcac aggaatgaaa accattagtg ataatttcat caccgaacag
     2641 aaggttatct cggtacccaa tcgccgccgc cgccgcctct tcctcctcct cgccctcgtc
     2701 gtcgtcgtct tcgtcgtaca agttgtcggc cagcggtggc tcccgagatg ggccctgcag
     2761 gcccggccca ttgtctcctt ccccagccgc cgcagtcgcc tgggcctctt gctccccgcc
     2821 tgccgccgcc gcctctgcct ccgcctcccg ccacagcgcc gccgccgccg cacccgggca
     2881 gcccctggcc gccgccggca cctcacgctc tggggccgcc ccaccgggct cgcccgggct
     2941 ccgctcgagg ccgggaccat ctctccgcgg cctcttgcgg agcggctccc cggcggggga
     3001 cgacgcggcc tccctgtcgg cccccgccgc cgagggggag ccgccgggct gaagggcgag
     3061 ggccgcctcg tccgccgaat tcccgggttt atcgtcatcg tccttgtagt ccatcctgag
     3121 gatccgagct cggtaccaag cttagatctc ctccaaaaaa gcctcttcac tacttctgga
     3181 atagctcaga ggccgaggcg gcctcggcct ctgcataaat aaaaaaaatt agtcagccat
     3241 ggggcggaga atgggcggaa ctgggcggag ttaggggcgg gatgggcgga gttaggggcg
     3301 ggactatggt tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc
     3361 ctggggactt tccttgctga ctaattgaga tgcatgcttt gcatacttct gcctgctggg
     3421 gagcctgggg actttccaca ccctaactga cacacattcc acagctggtt ctttccgcct
     3481 cagaaggtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     3541 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     3601 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     3661 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     3721 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     3781 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     3841 atgtggcgcg gtattatccc gtgttgacgc cgggcaagag caactcggtc gccgcataca
     3901 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     3961 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     4021 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     4081 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     4141 cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg ttgcgcaaac tattaactgg
     4201 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     4261 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     4321 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     4381 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     4441 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     4501 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     4561 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     4621 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     4681 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     4741 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct
     4801 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     4861 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     4921 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     4981 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     5041 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     5101 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     5161 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     5221 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     5281 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     5341 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     5401 agtgagcgag gaagcggaag
//

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