| 质粒名称: | PH-PLCD1-GFP |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 51407 |
| 存储实验室: | Tamas Balla |
|
载体骨架基本信息 |
|
| 载体名称: | pEGFP-N1 |
| 载体出品公司: | Clontech |
| 空载体大小: | -- |
| 完整质粒大小: | 5520 bp |
| 载体修饰: | -- |
| 载体类型: | 哺乳动物表达 |
| 筛选标记: | 新霉素（G418筛选） |
| 载体抗性: | 卡那霉素，50μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | PH-PLCD1 (aa 1–170) |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 510 bp |
| 基因突变: | -- |
| GenBank编号 | NP_006216.2 |
| 启动子: | -- |
| 标签/融合蛋白: | EGFP（插入物上的C端） |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | EcoRI(未破坏) |
| 3’克隆位点: | BamHI (未破坏) |
| 5’测序引物 | CMV-F |
| 3’测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | PH-PLCD1-GFP was a gift from Tamas Balla (Addgene plasmid # 51407 ; http://n2t.net/addgene:51407 ; RRID:Addgene_51407) |
| 参考文献部分: | Visualization of phosphoinositides that bind pleckstrin homology domains: calcium- and agonist-induced dynamic changes and relationship to myo-[3H]inositol-labeled phosphoinositide pools. Varnai P, Balla T. J Cell Biol. 1998 Oct 19;143(2):501-10. 10.1083/jcb.143.2.501 PubMed 9786958 |
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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 PH-PLCD1-GFP 5222 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Is a biosensor for PI(4,5)P2.
ACCESSION .
VERSION .
KEYWORDS PH-PLCD1-GFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5222)
AUTHORS Varnai P, Balla T
TITLE Visualization of phosphoinositides that bind pleckstrin homology
domains: calcium- and agonist-induced dynamic changes and
relationship to myo-[3H]inositol-labeled phosphoinositide pools.
JOURNAL J Cell Biol. 1998 Oct 19;143(2):501-10.
PUBMED 9786958
REFERENCE 2 (bases 1 to 5222)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..5222
/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"
regulatory 1162..1171
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 1168..1887
/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 1168..1884
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(1213..1234)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(1474..1493)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1821..1842
/label=EGFP-C
/note="EGFP, forward primer"
polyA_signal 2010..2131
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2047..2066)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 2101..2120
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(2138..2593)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2275..2296)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2487..2506
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 2620..2724
/gene="bla"
/label=AmpR promoter
promoter 2726..3083
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(2749..2769)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 2934..3069
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2996..3015
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3118..3912
/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(3172..3191)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3782..3801
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(4100..4119)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 4144..4191
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 4520..5108
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5009..5028
/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 caagcttcga attcacggca tggactcggg ccgggacttc
661 ctgaccctgc acggcctaca ggatgatgag gatctacagg cgctgctgaa gggcagccag
721 ctcctgaagg tgaagtccag ctcatggagg agagagcgct tctacaagtt gcaggaggac
781 tgcaagacca tctggcagga gtcccgcaag gtcatgcgga ccccggagtc ccagctgttc
841 tccatcgagg acattcagga ggtgcgaatg gggcaccgca cggagggtct ggagaagttc
901 gcccgtgatg tgcccgagga ccgctgcttc tccattgtct tcaaggacca gcgcaataca
961 ctagacctca tcgccccatc gccagctgat gcccagcact gggtgctggg gctgcacaag
1021 atcatccacc actcaggctc catggaccag cgtcagaagc tacagcactg gattcactcc
1081 tgcttgcgaa aagctgacaa aaacaaggac aacaagatga gcttcaagga gctgcagaac
1141 ttcctgaagg atccaccggt cgccaccatg gtgagcaagg gcgaggagct gttcaccggg
1201 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
1261 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc
1321 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc
1381 ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
1441 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
1501 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
1561 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc
1621 tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac
1681 atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
1741 ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac
1801 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact
1861 ctcggcatgg acgagctgta caagtaaagc ggccgcgact ctagatcata atcagccata
1921 ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga
1981 aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca
2041 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
2101 gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa ttgtaagcgt taatattttg
2161 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
2221 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
2281 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
2341 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
2401 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
2461 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
2521 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
2581 ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
2641 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
2701 caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc
2761 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
2821 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
2881 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
2941 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
3001 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
3061 ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg
3121 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
3181 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
3241 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa
3301 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
3361 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
3421 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
3481 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
3541 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
3601 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc
3661 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
3721 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
3781 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
3841 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
3901 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
3961 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
4021 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
4081 accctagggg gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta
4141 tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg
4201 gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat
4261 acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc
4321 tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca tatatacttt
4381 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
4441 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
4501 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
4561 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
4621 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
4681 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
4741 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
4801 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
4861 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
4921 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
4981 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
5041 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
5101 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
5161 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc
5221 at
//
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