| 质粒名称: | GFP-PH-FAPP1 |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 161986 |
| 存储实验室: | Tamas Balla, David Bryant |
|
载体骨架基本信息 |
|
| 载体名称: | eGFP-C1 |
| 完整质粒大小: | 5013bp |
| 载体类型: | 哺乳细胞表达 |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | NEB Stable |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | PH-FAPP1/PLEKHA3 |
| 物种来源: | H. sapiens (human |
| 基因突变: | PLEKHA3 (a.k.a. FAPP1) |
| GenBank编号 | JN247409 |
| 标签/融合蛋白: | GFP (N terminal on backbone) |
| 克隆方法: | 未知 |
| 5’测序引物未知 | 未知 |
|
文献引用方法 |
|
| 材料和方法部分: | GFP-PH-FAPP1 was a gift from Tamas Balla & David Bryant (Addgene plasmid # 161986 ; http://n2t.net/addgene:161986 ; RRID:Addgene_161986) |
| 参考文献部分: | A plasma membrane pool of phosphatidylinositol 4-phosphate is generated by phosphatidylinositol 4-kinase type-III alpha: studies with the PH domains of the oxysterol binding protein and FAPP1. Balla A, Tuymetova G, Tsiomenko A, Varnai P, Balla T. Mol Biol Cell. 2005 Mar;16(3):1282-95. doi: 10.1091/mbc.e04-07-0578. Epub 2005 Jan 5. 10.1091/mbc.e04-07-0578 PubMed 15635101 |
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载体基本信息
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载体简介
载体序列
LOCUS GFP-PH-FAPP1 5013 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Expresses GFP-tagged PH-FAPP1. PI(4)P biosensor.
ACCESSION .
VERSION .
KEYWORDS GFP-PH-FAPP1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5013)
AUTHORS Balla A, Tuymetova G, Tsiomenko A, Varnai P, Balla T
TITLE A plasma membrane pool of phosphatidylinositol 4-phosphate is
generated by phosphatidylinositol 4-kinase type-III alpha: studies
with the PH domains of the oxysterol binding protein and FAPP1.
JOURNAL Mol Biol Cell. 2005 Mar;16(3):1282-95. doi: 10.1091/mbc.e04-07-0578.
Epub 2005 Jan 5.
PUBMED 15635101
REFERENCE 2 (bases 1 to 5013)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..5013
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 91..394
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 395..598
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 549..569
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
regulatory 983..992
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 989..1708
/codon_start=1
/product="the original enhanced GFP (Yang et al., 1996)"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
CDS 989..1705
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(1034..1055)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(1295..1314)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1642..1663
/label=EGFP-C
/note="EGFP, forward primer"
polyA_signal 1831..1952
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1868..1887)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 1922..1941
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(1959..2414)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2096..2117)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2308..2327
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 2441..2545
/gene="bla"
/label=AmpR promoter
promoter 2547..2904
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(2570..2590)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 2755..2890
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2817..2836
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 2939..3733
/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(2993..3012)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3603..3622
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(3921..3940)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 3965..4012
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 4341..4929
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4830..4849
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 tctgtggata accgtattac cgccatgcat tagttattaa tagtaatcaa ttacggggtc
61 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
121 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
181 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
241 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
301 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
361 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa
421 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
481 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
541 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctgg
601 tttagtgaac cgtcagatcc gctagcgcta ccggactcag atctcgagct caagcttcga
661 attcaccatg gagggggtgt tgtacaagtg gaccaactat ctcacaggct ggcagcctcg
721 ttggtttgtt ttagataatg gaatcttatc ctactatgat tcacaagatg atgtttgcaa
781 agggagcaaa ggaagcataa agatggcagt ttgtgaaatt aaagttcatt cagcagacaa
841 cacaagaatg gaattaatca ttcctggaga gcagcatttc tacatgaagg cagtgaatgc
901 agctgaaaga cagaggtggc tggtcgctct ggggagctcc aaagcatgtt tgactgatac
961 aaggactaag gatccaccgg tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg
1021 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc
1081 cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac
1141 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg
1201 cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
1261 aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc
1321 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt
1381 caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt
1441 ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa
1501 catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga
1561 cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga
1621 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
1681 tctcggcatg gacgagctgt acaagtaaag cggccgcgac tctagatcat aatcagccat
1741 accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg
1801 aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac
1861 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
1921 tgtggtttgt ccaaactcat caatgtatct taaggcgtaa attgtaagcg ttaatatttt
1981 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
2041 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
2101 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
2161 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
2221 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
2281 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
2341 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
2401 gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
2461 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
2521 tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt
2581 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
2641 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
2701 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
2761 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
2821 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
2881 tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat
2941 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
3001 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
3061 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
3121 agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
3181 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
3241 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
3301 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
3361 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
3421 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg
3481 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
3541 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
3601 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
3661 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
3721 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg
3781 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
3841 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
3901 caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct
3961 atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc
4021 ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa
4081 tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg
4141 ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt
4201 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
4261 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
4321 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
4381 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
4441 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
4501 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
4561 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
4621 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
4681 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
4741 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
4801 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
4861 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
4921 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
4981 gctcacatgt tctttcctgc gttatcccct gat
//
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