| 质粒名称: | pEGFP-n1-APP |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 69924 |
| 存储实验室: | Zita Balklava, Thomas Wassmer |
|
载体骨架基本信息 |
|
| 载体名称: | pEGFP-N1 |
| 载体出品公司: | Clontech |
| 载体抗性: | 卡那霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | 6809 bp |
| 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: | APP,APP695 |
| 物种来源: | H. sapiens (human) |
| 插入大小: | -- |
| 基因突变: | - |
| GenBank编号 | -- |
| 启动子 | CMV |
| 标签/融合蛋白: | EGFP（主干上的C端） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | NheI (未破坏) |
| 3' 克隆位点: | SalI (未破坏) |
| 5' 测序引物 | CMV-F |
| 3' 测序引物 | EGFP-N |
|
文献引用方法 |
|
| 材料和方法部分: | pEGFP-n1-APP was a gift from Zita Balklava & Thomas Wassmer (Addgene plasmid # 69924 ; http://n2t.net/addgene:69924 ; RRID:Addgene_69924) |
| 参考文献部分: | APP controls the formation of PI(3,5)P vesicles through its binding of the PIKfyve complex. Currinn H, Guscott B, Balklava Z, Rothnie A, Wassmer T. Cell Mol Life Sci. 2015 Jul 28. 10.1007/s00018-015-1993-0 PubMed 26216398 |
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 pEGFP-n1-APP 6815 bp ds-DNA circular SYN 12-MAR-2019
DEFINITION mammalian expression of human APP fused to EGFP.
ACCESSION .
VERSION .
KEYWORDS pEGFP-n1-APP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6815)
AUTHORS Currinn H, Guscott B, Balklava Z, Rothnie A, Wassmer T
TITLE APP controls the formation of PI(3,5)P vesicles through its binding
of the PIKfyve complex.
JOURNAL Cell Mol Life Sci. 2015 Jul 28.
PUBMED 26216398
REFERENCE 2 (bases 1 to 6815)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Mar 12, 2019 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6815
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 61..364
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 365..568
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 519..539
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
CDS complement(961..972)
/codon_start=1
/gene="cspA"
/product="translation enhancing element for E. coli (Qing
et al., 2004)"
/label=TEE
/translation="NHKV"
CDS 1452..1472
/codon_start=1
/product="6xHis affinity tag"
/label=7xHis
/translation="HHHHHHH"
regulatory 2755..2764
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 2761..3480
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(2806..2827)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(3067..3086)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 3414..3435
/label=EGFP-C
/note="EGFP, forward primer"
polyA_signal 3603..3724
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3640..3659)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3694..3713
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(3731..4186)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3868..3889)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 4080..4099
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 4213..4317
/gene="bla"
/label=AmpR promoter
promoter 4319..4676
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(4342..4362)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 4527..4662
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 4589..4608
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 4711..5505
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(4765..4784)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 5375..5394
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(5693..5712)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 5737..5784
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 6113..6701
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6602..6621
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
601 ccggactcag atctcgagct caagcttgcc accatgctgc ccggtttggc actgctcctg
661 ctggccgcct ggacggctcg ggcgctggag gtacccactg atggtaatgc tggcctgctg
721 gctgaacccc agattgccat gttctgtggc agactgaaca tgcacatgaa tgtccagaat
781 gggaagtggg attcagatcc atcagggacc aaaacctgca ttgataccaa ggaaggcatc
841 ctgcagtatt gccaagaagt ctaccctgaa ctgcagatca ccaatgtggt agaagccaac
901 caaccagtga ccatccagaa ctggtgcaag cggggccgca agcagtgcaa gacccatccc
961 cactttgtga ttccctaccg ctgcttagtt ggtgagtttg taagtgatgc ccttctcgtt
1021 cctgacaagt gcaaattctt acaccaggag aggatggatg tttgcgaaac tcatcttcac
1081 tggcacaccg tcgccaaaga gacatgcagt gagaagagta ccaacttgca tgactacggc
1141 atgttgctgc cctgcggaat tgacaagttc cgaggggtag agtttgtgtg ttgcccactg
1201 gctgaagaaa gtgacaatgt ggattctgct gatgcggagg aggatgactc ggatgtctgg
1261 tggggcggag cagacacaga ctatgcagat gggagtgaag acaaagtagt agaagtagca
1321 gaggaggaag aagtggctga ggtggaagaa gaagaagccg atgatgacga ggacgatgag
1381 gatggtgatg aggtagagga agaggctgag gaaccctacg aagaagccac agagagaacc
1441 accagcattg ccaccaccac caccaccacc acagagtctg tggaagaggt ggttcgagtt
1501 cctacaacag cagccagtac ccctgatgcc gttgacaagt atctcgagac acctggggat
1561 gagaatgaac atgcccattt ccagaaagcc aaagagaggc ttgaggccaa gcaccgagag
1621 agaatgtccc aggtcatgag agaatgggaa gaggcagaac gtcaagcaaa gaacttgcct
1681 aaagctgata agaaggcagt tatccagcat ttccaggaga aagtggaatc tttggaacag
1741 gaagcagcca acgagagaca gcagctggtg gagacacaca tggccagagt ggaagccatg
1801 ctcaatgacc gccgccgcct ggccctggag aactacatca ccgctctgca ggctgttcct
1861 cctcggcctc gtcacgtgtt caatatgcta aagaagtatg tccgcgcaga acagaaggac
1921 agacagcaca ccctaaagca tttcgagcat gtgcgcatgg tggatcccaa gaaagccgct
1981 cagatccggt cccaggttat gacacacctc cgtgtgattt atgagcgcat gaatcagtct
2041 ctctccctgc tctacaacgt gcctgcagtg gccgaggaga ttcaggatga agttgatgag
2101 ctgcttcaga aagagcaaaa ctattcagat gacgtcttgg ccaacatgat tagtgaacca
2161 aggatcagtt acggaaacga tgctctcatg ccatctttga ccgaaacgaa aaccaccgtg
2221 gagctccttc ccgtgaatgg agagttcagc ctggacgatc tccagccgtg gcattctttt
2281 ggggctgact ctgtgccagc caacacagaa aacgaagttg agcctgttga tgcccgccct
2341 gctgccgacc gaggactgac cactcgacca ggttctgggt tgacaaatat caagacggag
2401 gagatctctg aagtgaagat ggatgcagaa ttccgacatg actcaggata tgaagttcat
2461 catcaaaaat tggtgttctt tgcagaagat gtgggttcaa acaaaggtgc aatcattgga
2521 ctcatggtgg gcggtgttgt catagcgaca gtgatcgtca tcaccttggt gatgctgaag
2581 aagaaacagt acacatccat tcatcatggt gtggtggagg ttgacgccgc tgtcacccca
2641 gaggagcgcc acctgtccaa gatgcagcag aacggctacg aaaatccaac ctacaagttc
2701 tttgagcaga tgcagaacgg gtcgacggta ccgcgggccc gggatccacc ggtcgccacc
2761 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
2821 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
2881 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
2941 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag
3001 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
3061 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
3121 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
3181 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
3241 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
3301 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
3361 tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc
3421 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa
3481 agcggccgcg actctagatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt
3541 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg
3601 ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
3661 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
3721 cttaaggcgt aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
3781 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
3841 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
3901 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
3961 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
4021 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
4081 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
4141 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt
4201 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
4261 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtcct
4321 gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
4381 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
4441 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
4501 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
4561 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
4621 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc
4681 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
4741 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
4801 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
4861 accgacctgt ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg
4921 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
4981 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
5041 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
5101 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
5161 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
5221 ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
5281 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
5341 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
5401 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
5461 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
5521 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
5581 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
5641 agcgcgggga tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac
5701 ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa
5761 cgcacggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg
5821 tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac
5881 cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc
5941 ctgccatagc ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat
6001 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
6061 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
6121 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
6181 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
6241 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
6301 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
6361 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
6421 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
6481 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
6541 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
6601 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
6661 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
6721 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
6781 ctgattctgt ggataaccgt attaccgcca tgcat
//
