| 质粒名称: | pRK5-EGFP-Tau |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 46904 |
| 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: | MAPT,Tau,microtubule-associated protein tau |
| 物种来源: | H. sapiens (human) |
| 插入大小: | -- |
| 基因突变: | 这个WT-htau结构包含缺少N末端序列（4R0N）的人类四个重复tau，并且包含外显子1、4和5、7和9-13、内含子13和外显子14 |
| 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 was a gift from Karen Ashe (Addgene plasmid # 46904 ; http://n2t.net/addgene:46904 ; RRID:Addgene_46904) |
| 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 6519 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION expresses EGFP tagged Tau in mammalian cells.
ACCESSION .
VERSION .
KEYWORDS pRK5-EGFP-Tau
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"
primer_bind 94..111
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 228..249
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 264..294
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 302..318
/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 307..329
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 326..342
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 326..342
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
enhancer 371..750
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 751..954
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 905..925
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 951..975
/label=LNCX
/note="Human CMV promoter, forward primer"
promoter 1185..1203
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind 1185..1202
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind 1216..1235
/label=pMT2-F
/note="Synthetic intron, forward primer"
regulatory 1279..1288
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 1285..2001
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSNQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(1330..1351)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(1591..1610)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1938..1959
/label=EGFP-C
/note="EGFP, forward primer"
regulatory 3176..3185
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
polyA_signal 3189..3323
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3226..3245)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3280..3299
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
promoter 3392..3749
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(3415..3435)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 3600..3735
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3662..3681
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
primer_bind complement(3769..3786)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(3769..3785)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3778..3800)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 3998..4453
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4085..4104)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 4295..4316
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(4470..4489)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 4589..4611
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(4649..4667)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 4735..4839
/gene="bla"
/label=AmpR promoter
CDS 4840..5700
/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(5058..5077)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 5871..6459
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6360..6379
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
61 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
121 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
181 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
241 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
301 attgtgagcg gataacaatt tcacacagga aacagctatg acatgattac gaattaattc
361 gagctcgccc gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta
421 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
481 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
541 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
601 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
661 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
721 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
781 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
841 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
901 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta
961 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac
1021 cgggaccgat ccagcctccg cggccgggaa cggtgcattg gaacgcggat tccccgtgcc
1081 aagagtgacg taagtaccgc ctatagagtc tataggccca cccccttggc ttcgttagaa
1141 cgcggctaca attaatacat aaccttatgt atcatacaca tacgatttag gtgacactat
1201 agaataacat ccactttgcc tttctctcca caggtgtcca ctcccaggtc caactgcacc
1261 tcggttctat cgatggtcgc caccatggtg agcaagggcg aggagctgtt caccggggtg
1321 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
1381 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
1441 aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc
1501 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
1561 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
1621 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
1681 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
1741 atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc
1801 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
1861 cccgtgctgc tgcccgacaa ccactacctg agcaaccagt ccgccctgag caaagacccc
1921 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
1981 ggcatggacg agctgtacaa gggatccgct gagccccgcc aggagttcga agtgatggaa
2041 gatcacgctg ggacgtacgg gttgggggac aggaaagatc aggggggcta caccatgcac
2101 caagaccaag agggtgacac ggacgctggc ctgaaagctg aagaagcagg cattggagac
2161 acccccagcc tggaagacga agctgctggt cacgtgaccc aagctcgcat ggtcagtaaa
2221 agcaaagacg ggactggaag cgatgacaaa aaagccaagg gggctgatgg taaaacgaag
2281 atcgccacac cgcggggagc agcccctcca ggccagaagg gccaggccaa cgccaccagg
2341 attccagcaa aaaccccgcc cgctccaaag acaccaccca gctctggtga acctccaaaa
2401 tcaggggatc gcagcggcta cagcagcccc ggctccccag gcactcccgg cagccgctcc
2461 cgcaccccgt cccttccaac cccacccacc cgggagccca agaaggtggc agtggtccgt
2521 actccaccca agtcgccgtc ttccgccaag agccgcctgc agacagcccc cgtgcccatg
2581 ccagacctga agaatgtcaa gtccaagatc ggctccactg agaacctgaa gcaccagccg
2641 ggaggcggga aggtgcagat aattaataag aagctggatc ttagcaacgt ccagtccaag
2701 tgtggctcaa aggataatat caaacacgtc ccgggaggcg gcagtgtgca aatagtctac
2761 aaaccagttg acctgagcaa ggtgacctcc aagtgtggct cattaggcaa catccatcat
2821 aaaccaggag gtggccaggt ggaagtaaaa tctgagaagc ttgacttcaa ggacagagtc
2881 cagtcgaaga ttgggtccct ggacaatatc acccacgtcc ctggcggagg aaataaaaag
2941 attgaaaccc acaagctgac cttccgcgag aacgccaaag ccaagacaga ccacggggcg
3001 gagatcgtgt acaagtcgcc agtggtgtct ggggacacgt ctccacggca tctcagcaat
3061 gtctcctcca ccggcagcat cgacatggta gactcgcccc agctcgccac gctagctgac
3121 gaggtgtctg cctccctggc caagcagggt ttgtgagtcg acctgcagaa gcttggccgc
3181 catggcccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
3241 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
3301 atgtatctta tcatgtctgg atcgggaatt aattcggcgc agcaccatgg cctgaaataa
3361 cctctgaaag aggaacttgg ttaggtacct tctgaggcgg aaagaaccag ctgtggaatg
3421 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
3481 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
3541 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
3601 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
3661 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
3721 gcttttttgg aggcctaggc ttttgcaaaa agctgttaac agcttggcac tggccgtcgt
3781 tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca
3841 tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
3901 gttgcgcagc ctgaatggcg aatggcgcct gatgcggtat tttctcctta cgcatctgtg
3961 cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta
4021 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
4081 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
4141 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
4201 aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt
4261 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
4321 acactcaacc ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggcc
4381 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
4441 acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
4501 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
4561 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
4621 tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat
4681 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
4741 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
4801 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
4861 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
4921 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
4981 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
5041 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
5101 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
5161 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
5221 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
5281 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
5341 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
5401 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
5461 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
5521 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
5581 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
5641 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
5701 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
5761 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
5821 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
5881 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
5941 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
6001 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
6061 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
6121 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
6181 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
6241 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
6301 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
6361 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
6421 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
6481 ttacggttcc tggccttttg ctggcctttt gctcacatg
//
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