| 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 |
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 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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