| 质粒名称: | Flag-SIRT1 H363Y,pECE-FLAG-SIRT1-H363Y |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 1792 |
| 存储实验室: | Michael Greenberg |
|
载体骨架基本信息 |
|
| 载体名称: | pECE |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 2900 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | -- |
| 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,Sir2 |
| 物种来源: | H. sapiens (human) |
| 插入大小: | -- |
| 基因突变: | H363Y, deacetylase domain mutation |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | Flag （插入N端子） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | HindIII（未销毁） |
| 3' 克隆位点: | XbaI（未销毁） |
| 5' 测序引物 | SV40pro-F |
| 3' 测序引物 | SV40pA-R |
|
文献引用方法 |
|
| 材料和方法部分: | Flag-SIRT1 H363Y was a gift from Michael Greenberg (Addgene plasmid # 1792 ; http://n2t.net/addgene:1792 ; RRID:Addgene_1792) |
| 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 |
买家导航
载体基本信息
载体图谱质粒图谱和多克隆位点信息
载体简介
载体序列
LOCUS Flag-SIRT1 H363Y 5420 bp ds-DNA circular SYN 06-AUG-2018
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
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
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(4587..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"
ORIGIN
1 aggcaaaggt tctctattaa ttaatatctg aggcacttca tggggtatgg aacttggaat
61 tagtgctact ggtcttactt tgagggaaga cccaataaca atgaggaggt caacttcatc
121 tttgtcatac ttcatggctc tatgaaactg ttctggtaaa ttttcaccaa aaaacacaat
181 ctctggtttc atgatagcaa gcggttcatc agctgggcac ctaggacatc gaggaactac
241 ctgattaaaa atatctcctc gtacagcttc acagtcaact ttgtatttac aaatcaggca
301 agatgctgtt gcaaaggaac cataacactg aattatcctt tggattcccg caacctgttc
361 cagcgtgtct atgttctggg tatagttgcg aagtagtttt ccttccttat ctgacaaggc
421 tatgaatttg tgacagagag atggctggaa ttgtccagga tatatttcct ttgcaaactt
481 gaagaatggt cttggatctt ttctgaaata ttcaatatca aacatcgctt gaggatctgg
541 aagatctggg aagtctacag caaggcgagc ataaatacca tcccttgacc tgaagtcagg
601 tattccacat gaaacagaca ccccagctcc agttagaact ataatttttt tgcactcttg
661 cagtaatttc acagcatctt caattgtatt aatatctttt ctttttttcc tttttggtgg
721 ttctgaaagg atattaataa caatctgcca cagtgtcata tcatccaact caggtggagg
781 tattgtttcc ggcaataaat ctttaagaat tgttcgagga tctgtgccaa tcataagatg
841 ttgctgaaca aaagtatatg gacctatccg tggccttgga gtccagtcac tagagcttgc
901 atgtgaggct ctatcctcct catcactttc acaggaatga aaaccattag tgataatttc
961 atcaccgaac agaaggttat ctcggtaccc aatcgccgcc gccgccgcct cttcctcctc
1021 ctcgccctcg tcgtcgtcgt cttcgtcgta caagttgtcg gccagcggtg gctcccgaga
1081 tgggccctgc aggcccggcc cattgtctcc ttccccagcc gccgcagtcg cctgggcctc
1141 ttgctccccg cctgccgccg ccgcctctgc ctccgcctcc cgccacagcg ccgccgccgc
1201 cgcacccggg cagcccctgg ccgccgccgg cacctcacgc tctggggccg ccccaccggg
1261 ctcgcccggg ctccgctcga ggccgggacc atctctccgc ggcctcttgc ggagcggctc
1321 cccggcgggg gacgacgcgg cctccctgtc ggcccccgcc gccgaggggg agccgccggg
1381 ctgaagggcg agggccgcct cgtccgccga attcccgggt ttatcgtcat cgtccttgta
1441 gtccatcctg aggatccgag ctcggtacca agcttagatc tcctccaaaa aagcctcttc
1501 actacttctg gaatagctca gaggccgagg cggcctcggc ctctgcataa ataaaaaaaa
1561 ttagtcagcc atggggcgga gaatgggcgg aactgggcgg agttaggggc gggatgggcg
1621 gagttagggg cgggactatg gttgctgact aattgagatg catgctttgc atacttctgc
1681 ctgctgggga gcctggggac tttccttgct gactaattga gatgcatgct ttgcatactt
1741 ctgcctgctg gggagcctgg ggactttcca caccctaact gacacacatt ccacagctgg
1801 ttctttccgc ctcagaaggt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
1861 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
1921 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
1981 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
2041 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
2101 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
2161 taaagttctg ctatgtggcg cggtattatc ccgtgttgac gccgggcaag agcaactcgg
2221 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
2281 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
2341 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
2401 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
2461 cataccaaac gacgagcgtg acaccacgat gcctgcagca atggcaacaa cgttgcgcaa
2521 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
2581 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
2641 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
2701 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
2761 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
2821 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
2881 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
2941 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
3001 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
3061 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
3121 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
3181 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
3241 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
3301 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
3361 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
3421 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
3481 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
3541 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
3601 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
3661 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
3721 gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt ttctccttac
3781 gcatctgtgc ggtatttcac accgcatatc gacgctctcc cttatgcgac tcctgcatta
3841 ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc cgccgcaagg aatggtgcat
3901 gcaaggagat ggcgcccaac agtcccccgg ccacggggcc tgccaccata cccacgccga
3961 aacaagcgct catgagcccg aagtggcgag cccgatcttc cccatcggtg atgtcggcga
4021 tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt
4081 agaggatcca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca
4141 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
4201 aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg
4261 ggaggtgtgg gaggtttttt aaagcaagta aaacctctac agatgtgata tggctgatta
4321 tgatcattac ttatctaggt ataggctgcg caactgttgg gaagggcgat cggtgcgggc
4381 ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt
4441 aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgaat tgtaatacga
4501 ctcactatag ggcgaattga atttagcggc cgcgaattcg cccttgtcta gagtggaaca
4561 attcctgtac ctgcacaatt attacactat gatttgtttg atggatagtt catgtctgtt
4621 acttcctgtt tcacagatat agcttcatta attgcctctt gatcatctcc atcagtccca
4681 aatccagctc ctccagctct ctctggaaca tcaggctcat cttctaagcc attgtagaat
4741 tcttcaattt cactttcatc ctccatgggt tcttctaaac ttggactctg gcatgtccca
4801 ctatcactgt tactgccaca agaactagag gataagacgt catcttcaga gtctgaatat
4861 acctcagcgc catggaaaat gtaacgattt ggtggcaaaa acagatactg attaccatca
4921 agccgcctac taatctgctc ctttgccact ctattaggcc agcattttct cactgttcca
4981 gccactgaag ttctttcatt tttctcccca gtactagaac caacattctt caaatccgga
5041 ttttccatct gttcagcaat actttcaaca ttcctagaag tttgtacttc ctgtggtttt
5101 tcttccatac aaccttttga ttcagacaca tctaaatcat cattactctt agctgcttgg
5161 tctaaaagtg tgacaatcac tgaagaatct ggtggtgaag ttctttctgg tgaacttgag
5221 tcttctgaaa catgaagagg tgtgggtggc aactctgaca aataagccaa ttctttttgt
5281 gttcgtggag gtttttcagt aatttctgaa agctttacag ggttacagca aagtttggca
5341 tattcaccac ctaacctatg acacaattca ttaattatga catcacagtc tccaagaagc
5401 tctacatcaa aatgcagatg
//
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