| 质粒名称: | AAV.CBA.eGFP.2A.P301L-Tau |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 140425 |
| 存储实验室: | Bradley Hyman |
|
载体骨架基本信息 |
|
| 载体名称: | AAV.CBA.WPRE |
| 载体出品公司: | Bradley Hyman |
| 空载体大小: | 8570 bp |
| 完整质粒大小: | 6468 bp |
| 载体类型: | AAV |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | NEB Stable |
| 拷贝数: | 未知 |
|
插入基因信息 |
|
| 插入基因名称: | GFP-2A-Tau(P301L) |
| 物种来源: | H. sapiens (human), Synthetic |
| 基因突变: | changed Proline 301 to Leucine |
| GenBank编号 | NM_005910.5 |
| 启动子: | CBA |
| 标签/融合蛋白: | eGFP (N terminal on insert) |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | Xho1 (unknown if destroyed) |
| 3’克隆位点: | EcoRV (unknown if destroyed) |
| 5’测序引物 | primerdo not have |
| 3’测序引物 | primerdo not have |
|
文献引用方法 |
|
| 材料和方法部分: |
AAV.CBA.eGFP.2A.P301L-Tau was a gift from Bradley Hyman (Addgene plasmid # 140425 ; http://n2t.net/addgene:140425 ; RRID:Addgene_140425)
|
| 参考文献部分: | Experimental evidence for the age dependence of tau protein spread in the brain. Wegmann S, Bennett RE, Delorme L, Robbins AB, Hu M, McKenzie D, Kirk MJ, Schiantarelli J, Tunio N, Amaral AC, Fan Z, Nicholls S, Hudry E, Hyman BT. Sci Adv. 2019 Jun 26;5(6):eaaw6404. doi: 10.1126/sciadv.aaw6404. eCollection 2019 Jun. 10.1126/sciadv.aaw6404 PubMed 31249873 |
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 AAV.CBA.eGFP.2A.P301L-Tau 8568 bp ds-DNA circular SYN 27-MAR-2024
DEFINITION AAV.CBA.eGFP.2A.P301L-Tau propagation reporter P301L-2N4R-hTau.
ACCESSION .
VERSION .
KEYWORDS AAV.CBA.eGFP.2A.P301L-Tau
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8568)
AUTHORS Wegmann S, Bennett RE, Delorme L, Robbins AB, Hu M, McKenzie D, Kirk
MJ, Schiantarelli J, Tunio N, Amaral AC, Fan Z, Nicholls S, Hudry E,
Hyman BT
TITLE Experimental evidence for the age dependence of tau protein spread
in the brain.
JOURNAL Sci Adv. 2019 Jun 26;5(6):eaaw6404. doi: 10.1126/sciadv.aaw6404.
eCollection 2019 Jun.
PUBMED 31249873
REFERENCE 2 (bases 1 to 8568)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Mar 27, 2024 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..8568
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(102..121)
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
enhancer 188..567
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 569..848
/label=chicken beta-actin promoter
primer_bind 1261..1283
/label=pCAGGS-5
/note="Chimeric intron in CAG promoter, forward primer"
primer_bind 1345..1364
/label=pCAG-F
/note="Rabbit beta-globin intron, for pCAG plasmids,
forward primer"
misc_feature 1390..1442
/label=MCS
/note="multiple cloning site"
regulatory 1450..1459
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 1456..2172
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
CDS 1456..2172
/codon_start=1
/product="the original enhanced GFP (Yang et al., 1996)"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(1501..1522)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(1762..1781)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 2109..2130
/label=EGFP-C
/note="EGFP, forward primer"
CDS 2182..2235
/codon_start=1
/product="2A peptide from Thosea asigna virus capsid
protein"
/label=T2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="EGRGSLLTCGDVEENPGP"
misc_feature 3586..4174
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(3639..3659)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(4057..4068)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
primer_bind complement(4266..4285)
/label=Bglob-pA-R
/note="Rabbit beta-globin polyA region, reverse primer"
polyA_signal 4331..4386
/label=-beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal (Gil and
Proudfoot, 1987)"
primer_bind complement(4385..4404)
/label=rbglobpA-R
/note="Rabbit beta-globin polyA, reverse primer. Also
called rb-glob-pA-term-R"
repeat_region 4786..4915
/label=AAV2 ITR
rep_origin 5286..5799
/direction=RIGHT
/label=M13 ori
/note="M13 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(5373..5392)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 5583..5604
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(6281..6300)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 6400..6422
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(6460..6478)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 6546..6650
/gene="bla"
/label=AmpR promoter
CDS 6651..7511
/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(6869..6888)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 7682..8270
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 8171..8190
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 8424..8441
/label=L4440
/note="L4440 vector, forward primer"
ORIGIN
1 cgagcgcgca gagagggagt ggccaactcc atcactaggg gttccttgta gttaatgatt
61 aacccgccat gctacttatc tacgtagcca tgctctagcg gcctcggcct ctgcataaat
121 aaaaaaaatt agtcagccat gagcttggcc cattgcatac gttgtatcca tatcataata
181 tgtactcgac attgattatt gactagttat taatagtaat caattacggg gtcattagtt
241 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
301 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
361 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
421 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
481 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
541 tacgtattag tcatcgctat taccatggtc gaggtgagcc ccacgttctg cttcactctc
601 cccatctccc ccccctcccc acccccaatt ttgtatttat ttatttttta attattttgt
661 gcagcgatgg gggcgggggg gggggggggg ggcgcgcgcc aggcggggcg gggcggggcg
721 aggggcgggg cggggcgagg cggagaggtg cggcggcagc caatcagagc ggcgcgctcc
781 gaaagtttcc ttttatggcg aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc
841 ggcgggcggg agtcgctgcg cgctgccttc gccccgtgcc ccgctccgcc gccgcctcgc
901 gccgcccgcc ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc
961 cttctcctcc gggctgtaat tagcgcttgg tttaatgacg gcttgtttct tttctgtggc
1021 tgcgtgaaag ccttgagggg ctccgggagg gccctttgtg cggggggagc ggctcggggg
1081 gtgcgtgcgt gtgtgtgtgc gtggggagcg ccgcgtgcgg ctccgcgctg cccggcggct
1141 gtgagcgctg cgggcgcggc gcggggcttt gtgcgctccg cagtgtgcgc gaggggagcg
1201 cggccggggg cggtgccccg cggggggacg gctgccttcg ggggggacgg ggcagggcgg
1261 ggttcggctt ctggcgtgtg accggcggct ctagagcctc tgctaaccat gttcatgcct
1321 tcttcttttt cctacagctc ctgggcaacg tgctggttat tgtgctgtct catcattttg
1381 gcaaagaatc tcgagctcaa gcttcgaatt ctgcagtcga cggtaccgcg ggcccgggat
1441 ccaccggtcg ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
1501 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
1561 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
1621 gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac
1681 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
1741 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
1801 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
1861 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc
1921 gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc
1981 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
2041 ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag
2101 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
2161 gagctgtaca agggcagtgg agagggcaga ggaagtctgc taacatgcgg tgacgtcgag
2221 gagaatcctg gcccagctga gccccgccag gagttcgaag tgatggaaga tcacgctggg
2281 acgtacgggt tgggggacag gaaagatcag gggggctaca ccatgcacca agaccaagag
2341 ggtgacacgg acgctggcct gaaagaatct cccctgcaga cccccactga ggacggatct
2401 gaggaaccgg gctctgaaac ctctgatgct aagagcactc caacagcgga agatgtgaca
2461 gcacccttag tggatgaggg agctcccggc aagcaggctg ccgcgcagcc ccacacggag
2521 atcccagaag gaaccacagc tgaagaagca ggcattggag acacccccag cctggaagac
2581 gaagctgctg gtcacgtgac ccaagctcgc atggtcagta aaagcaaaga cgggactgga
2641 agcgatgaca aaaaagccaa gggggctgat ggtaaaacga agatcgccac accgcgggga
2701 gcagcccctc caggccagaa gggccaggcc aacgccacca ggattccagc aaaaaccccg
2761 cccgctccaa agacaccacc cagctctggt gaacctccaa aatcagggga tcgcagcggc
2821 tacagcagcc ccggctcccc aggcactccc ggcagccgct cccgcacccc gtcccttcca
2881 accccaccca cccgggagcc caagaaggtg gcagtggtcc gtactccacc caagtcgccg
2941 tcttccgcca agagccgcct gcagacagcc cccgtgccca tgccagacct gaagaatgtc
3001 aagtccaaga tcggctccac tgagaacctg aagcaccagc cgggaggcgg gaaggtgcag
3061 ataattaata agaagctgga tcttagcaac gtccagtcca agtgtggctc aaaggataat
3121 atcaaacacg tcctgggagg cggcagtgtg caaatagtct acaaaccagt tgacctgagc
3181 aaggtgacct ccaagtgtgg ctcattaggc aacatccatc ataaaccagg aggtggccag
3241 gtggaagtaa aatctgagaa gcttgacttc aaggacagag tccagtcgaa gattgggtcc
3301 ctggacaata tcacccacgt ccctggcgga ggaaataaaa agattgaaac ccacaagctg
3361 accttccgcg agaacgccaa agccaagaca gaccacgggg cggagatcgt gtacaagtcg
3421 ccagtggtgt ctggggacac gtctccacgg catctcagca atgtctcctc caccggcagc
3481 atcgacatgg tagactcgcc ccagctcgcc acgctagctg acgaggtgtc tgcctccctg
3541 gccaagcagg gtttgtagac tagcgatatc ggtaccggat ccgataatca acctctggat
3601 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
3661 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
3721 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg
3781 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc
3841 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa
3901 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
3961 tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc
4021 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt
4081 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag
4141 acgagtcgga tctccctttg ggccgcctcc ccgcatcgga tccaattcac tcctcaggtg
4201 caggctgcct atcagaaggt ggtggctggt gtggccaatg ccctggctca caaataccac
4261 tgagatcttt ttccctctgc caaaaattat ggggacatca tgaagcccct tgagcatctg
4321 acttctggct aataaaggaa atttattttc attgcaatag tgtgttggaa ttttttgtgt
4381 ctctcactcg gaaggacata tgggagggca aatcatttaa aacatcagaa tgagtatttg
4441 gtttagagtt tggcaacata tgcccatatg ctggctgcca tgaacaaagg ttggctataa
4501 agaggtcatc agtatatgaa acagccccct gctgtccatt ccttattcca tagaaaagcc
4561 ttgacttgag gttagatttt ttttatattt tgttttgtgt tatttttttc tttaacatcc
4621 ctaaaatttt ccttacatgt tttactagcc agatttttcc tcctctcctg actactccca
4681 gtcatagctg tccctcttct cttatggaga tccctcgacc tgcagcccaa gctctagagc
4741 atggctacgt agataagtag catggcgggt taatcattaa ctacaaggaa cccctagtga
4801 tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg
4861 tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctggc
4921 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
4981 aatggcgatt ccgttgcaat ggctggcggt aatattgttc tggatattac cagcaaggcc
5041 gatagtttga gttcttctac tcaggcaagt gatgttatta ctaatcaaag aagtattgcg
5101 acaacggtta atttgcgtga tggacagact cttttactcg gtggcctcac tgattataaa
5161 aacacttctc aggattctgg cgtaccgttc ctgtctaaaa tccctttaat cggcctcctg
5221 tttagctccc gctctgattc taacgaggaa agcacgttat acgtgctcgt caaagcaacc
5281 atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
5341 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
5401 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
5461 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
5521 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
5581 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
5641 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
5701 atttaacgcg aattttaaca aaatattaac gcttacaatt taaatatttg cttatacaat
5761 cttcctgttt ttggggcttt tctgattatc aaccggggta catatgattg acatgctagt
5821 tttacgatta ccgttcatcg attctcttgt ttgctccaga ctctcaggca atgacctgat
5881 agcctttgta gagacctctc aaaaatagct accctctccg gcatgaattt atcagctaga
5941 acggttgaat atcatattga tggtgatttg actgtctccg gcctttctca cccgtttgaa
6001 tctttaccta cacattactc aggcattgca tttaaaatat atgagggttc taaaaatttt
6061 tatccttgcg ttgaaataaa ggcttctccc gcaaaagtat tacagggtca taatgttttt
6121 ggtacaaccg atttagcttt atgctctgag gctttattgc ttaattttgc taattctttg
6181 ccttgcctgt atgatttatt ggatgttgga atcgcctgat gcggtatttt ctccttacgc
6241 atctgtgcgg tatttcacac cgcatatggt gcactctcag tacaatctgc tctgatgccg
6301 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
6361 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
6421 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt
6481 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
6541 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
6601 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
6661 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
6721 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
6781 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
6841 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
6901 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
6961 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
7021 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
7081 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
7141 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
7201 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
7261 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
7321 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
7381 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
7441 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
7501 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
7561 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
7621 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
7681 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
7741 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
7801 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
7861 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
7921 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
7981 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
8041 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
8101 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
8161 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
8221 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
8281 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
8341 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
8401 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
8461 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgc gcgctcgctc
8521 gctcactgag gccgcccggg caaagcccgg gcggcctcag tgagcgag
//
