| 质粒名称: | pRK5-EGFP-Tau P301L |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 46908 |
| 存储实验室: | Karen Ashe |
|
载体骨架基本信息 |
|
| 载体名称: | pRK5 |
| 载体出品公司: | Clontech |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | -- |
| 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: | -- |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | Tau P301L,Tau,microtubule-associated protein tau,MAPT |
| 物种来源: | H. sapiens (human) |
| 插入大小: | -- |
| 基因突变: | P301L |
| GenBank编号 | -- |
| 启动子 | CMV |
| 标签/融合蛋白: | EGFP（插入的N端） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | Cla1 (未破坏) |
| 3' 克隆位点: | SalI (未破坏) |
| 5' 测序引物 | SP6 |
| 3' 测序引物 | SV40pA-R |
|
文献引用方法 |
|
| 材料和方法部分: | pRK5-EGFP-Tau P301L was a gift from Karen Ashe (Addgene plasmid # 46908 ; http://n2t.net/addgene:46908 ; RRID:Addgene_46908) |
| 参考文献部分: | Tau mislocalization to dendritic spines mediates synaptic dysfunction independently of neurodegeneration. Hoover BR, Reed MN, Su J, Penrod RD, Kotilinek LA, Grant MK, Pitstick R, Carlson GA, Lanier LM, Yuan LL, Ashe KH, Liao D. Neuron. 2010 Dec 22;68(6):1067-81. doi: 10.1016/j.neuron.2010.11.030. 10.1016/j.neuron.2010.11.030 PubMed 21172610 |
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 pRK5-EGFP-Tau P301L 6519 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION expresses EGFP tagged Tau P301L in mammalian cells.
ACCESSION .
VERSION .
KEYWORDS pRK5-EGFP-Tau P301L
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6519)
AUTHORS Hoover BR, Reed MN, Su J, Penrod RD, Kotilinek LA, Grant MK,
Pitstick R, Carlson GA, Lanier LM, Yuan LL, Ashe KH, Liao D
TITLE Tau mislocalization to dendritic spines mediates synaptic
dysfunction independently of neurodegeneration.
JOURNAL Neuron. 2010 Dec 22;68(6):1067-81. doi:
10.1016/j.neuron.2010.11.030.
PUBMED 21172610
REFERENCE 2 (bases 1 to 6519)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6519
/organism="synthetic DNA construct"
/mol_type="other DNA"
protein_bind 107..128
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 143..173
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 181..197
/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 186..208
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 205..221
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 205..221
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
enhancer 250..629
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 630..833
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 784..804
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 830..854
/label=LNCX
/note="Human CMV promoter, forward primer"
promoter 1064..1082
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind 1064..1081
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind 1095..1114
/label=pMT2-F
/note="Synthetic intron, forward primer"
regulatory 1158..1167
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 1164..1880
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(1209..1230)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(1470..1489)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1817..1838
/label=EGFP-C
/note="EGFP, forward primer"
regulatory 3055..3064
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
polyA_signal 3068..3202
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3105..3124)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3159..3178
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
promoter 3271..3628
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(3294..3314)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 3479..3614
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3541..3560
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
primer_bind complement(3648..3665)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(3648..3664)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3657..3679)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 3877..4332
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3964..3983)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 4174..4195
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(4349..4368)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 4468..4490
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(4528..4546)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 4614..4718
/gene="bla"
/label=AmpR promoter
CDS 4719..5579
/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(4937..4956)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 5750..6338
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6239..6258
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 6492..6509
/label=L4440
/note="L4440 vector, forward primer"
ORIGIN
1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
181 ttgtgagcgg ataacaattt cacacaggaa acagctatga catgattacg aattaattcg
241 agctcgcccg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag
301 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
361 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
421 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
481 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
541 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
601 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
661 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
721 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
781 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctcgtttag
841 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagacacc
901 gggaccgatc cagcctccgc ggccgggaac ggtgcattgg aacgcggatt ccccgtgcca
961 agagtgacgt aagtaccgcc tatagagtct ataggcccac ccccttggct tcgttagaac
1021 gcggctacaa ttaatacata accttatgta tcatacacat acgatttagg tgacactata
1081 gaataacatc cactttgcct ttctctccac aggtgtccac tcccaggtcc aactgcacct
1141 cggttctatc gatggtcgcc accatggtga gcaagggcga ggagctgttc accggggtgg
1201 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg
1261 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca
1321 agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca
1381 gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
1441 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
1501 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
1561 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata
1621 tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg
1681 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
1741 ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca
1801 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
1861 gcatggacga gctgtacaag ggatccgctg agccccgcca ggagttcgaa gtgatggaag
1921 atcacgctgg gacgtacggg ttgggggaca ggaaagatca ggggggctac accatgcacc
1981 aagaccaaga gggtgacacg gacgctggcc tgaaagctga agaagcaggc attggagaca
2041 cccccagcct ggaagacgaa gctgctggtc acgtgaccca agctcgcatg gtcagtaaaa
2101 gcaaagacgg gactggaagc gatgacaaaa aagccaaggg ggctgatggt aaaacgaaga
2161 tcgccacacc gcggggagca gcccctccag gccagaaggg ccaggccaac gccaccagga
2221 ttccagcaaa aaccccgccc gctccaaaga caccacccag ctctggtgaa cctccaaaat
2281 caggggatcg cagcggctac agcagccccg gctccccagg cactcccggc agccgctccc
2341 gcaccccgtc ccttccaacc ccacccaccc gggagcccaa gaaggtggca gtggtccgta
2401 ctccacccaa gtcgccgtct tccgccaaga gccgcctgca gacagccccc gtgcccatgc
2461 cagacctgaa gaatgtcaag tccaagatcg gctccactga gaacctgaag caccagccgg
2521 gaggcgggaa ggtgcagata attaataaga agctggatct tagcaacgtc cagtccaagt
2581 gtggctcaaa ggataatatc aaacacgtcc tgggaggcgg cagtgtgcaa atagtctaca
2641 aaccagttga cctgagcaag gtgacctcca agtgtggctc attaggcaac atccatcata
2701 aaccaggagg tggccaggtg gaagtaaaat ctgagaagct tgacttcaag gacagagtcc
2761 agtcgaagat tgggtccctg gacaatatca cccacgtccc tggcggagga aataaaaaga
2821 ttgaaaccca caagctgacc ttccgcgaga acgccaaagc caagacagac cacggggcgg
2881 agatcgtgta caagtcgcca gtggtgtctg gggacacgtc tccacggcat ctcagcaatg
2941 tctcctccac cggcagcatc gacatggtag actcgcccca gctcgccacg ctagctgacg
3001 aggtgtctgc ctccctggcc aagcagggtt tgtgagtcga cctgcagaag cttggccgcc
3061 atggcccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
3121 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
3181 tgtatcttat catgtctgga tcgggaatta attcggcgca gcaccatggc ctgaaataac
3241 ctctgaaaga ggaacttggt taggtacctt ctgaggcgga aagaaccagc tgtggaatgt
3301 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
3361 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
3421 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
3481 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
3541 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
3601 cttttttgga ggcctaggct tttgcaaaaa gctgttaaca gcttggcact ggccgtcgtt
3661 ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat
3721 ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
3781 ttgcgcagcc tgaatggcga atggcgcctg atgcggtatt ttctccttac gcatctgtgc
3841 ggtatttcac accgcatacg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa
3901 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
3961 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
4021 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
4081 aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
4141 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
4201 cactcaaccc tatctcgggc tattcttttg atttataagg gattttgccg atttcggcct
4261 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
4321 cgtttacaat tttatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
4381 agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
4441 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
4501 catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
4561 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
4621 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
4681 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
4741 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
4801 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
4861 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
4921 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
4981 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
5041 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
5101 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
5161 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
5221 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
5281 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
5341 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
5401 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
5461 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
5521 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
5581 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
5641 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
5701 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
5761 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
5821 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
5881 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
5941 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
6001 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
6061 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
6121 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
6181 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
6241 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
6301 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
6361 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
6421 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
6481 cgaccgagcg cagcgagtca gtgagcgagg aagcggaag
//
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