| 质粒名称: | GFP-C1-PLCdelta-PH |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 21179 |
| 存储实验室: | Vladislav Verkhusha |
|
载体骨架基本信息 |
|
| 载体名称: | C1 |
| 载体出品公司: | Clontech |
| 空载体大小: | 4700bp |
| 完整质粒大小: | 5253bp |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 新霉素（G418） |
| 载体抗性: | Kanamycin, 50 μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 未知 |
|
插入基因信息 |
|
| 插入基因名称: | PLCdelta |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 529bp |
| 基因突变: | PH domain of human PLC delta, amino acids Met1–Ile175. (Lab plasmid WC 0003) |
| 启动子: | CMV |
| 标签/融合蛋白: | GFP (N terminal on insert) |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | BamHI (unknown if destroyed) |
| 3’克隆位点: | XbaI (unknown if destroyed) |
| 5’测序引物 | n/a |
|
文献引用方法 |
|
| 材料和方法部分: |
GFP-C1-PLCdelta-PH was a gift from Tobias Meyer (Addgene plasmid # 21179 ; http://n2t.net/addgene:21179 ; RRID:Addgene_21179)
|
| 参考文献部分: | Receptor-induced transient reduction in plasma membrane PtdIns(4,5)P2 concentration monitored in living cells. Stauffer TP, Ahn S, Meyer T. Curr Biol. 1998 Mar 12. 8(6):343-6. 10.1016/S0960-9822(98)70135-6 PubMed 9512420 |
买家导航
GFP-C1-PLCdelta-PH载体质粒基本信息
GFP-C1-PLCdelta-PH质粒图谱载体图谱和GFP-C1-PLCdelta-PH载体序列质粒序列多克隆位点信息
GFP-C1-PLCdelta-PH质粒载体简介
GFP-C1-PLCdelta-PH质粒序列载体序列
LOCUS GFP-C1-PLCdelta-PH 5253 bp DNA circular SYN 14-APR-2025
DEFINITION encodes a PI(4,5)P2 lipid selective PH domain that can be used as a
fluorescent translocation biosensor to monitor changes or local
differences in the concentration of plasma membrane PI(4,5)P2
lipids.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5253)
AUTHORS Stauffer TP, Ahn S, Meyer T
TITLE Receptor-induced transient reduction in plasma membrane
PtdIns(4,5)P2 concentration monitored in living cells.
JOURNAL Curr Biol. 1998 Mar 12. 8(6):343-6.
PUBMED 9512420
REFERENCE 2 (bases 1 to 5253)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 14, 2025 from SnapGene Server 8.0.1
https://www.biofeng.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Curr Biol.
1998 Mar 12. 8(6):343-6."
COMMENT Sequence Label: GFP-C1-PLCdelta-PH
FEATURES Location/Qualifiers
source 1..5253
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 115..418
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 419..622
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 573..593
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
regulatory 661..670
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS join(667..669,670..672,673..1383)
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
CDS join(667..669,670..672,673..1383)
/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(712..733)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(973..992)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1320..1341
/label=EGFP-C
/note="EGFP, forward primer"
misc_feature 1384..1449
/label=MCS
/note="multiple cloning site"
polyA_signal 2095..2216
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2132..2151)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 2186..2205
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(2223..2678)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2360..2381)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2572..2591
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 2705..2809
/gene="bla"
/label=AmpR promoter
promoter 2811..3168
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
enhancer 2811..3002
/label=SV40 enhancer
/note="enhancer for the SV40 early promoter (Herr, 1993)"
primer_bind complement(2834..2854)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 3019..3154
/label=SV40 ori
/note="SV40 origin of replication"
CDS 3203..3997
/codon_start=1
/product="aminoglycoside phosphotransferase"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
CDS 3203..3997
/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(3257..3276)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3867..3886
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(4185..4204)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 4229..4276
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 4605..5193
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5094..5113
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat gcattagtta
61 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
121 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
181 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
241 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
301 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
361 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
421 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
481 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
541 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
601 ggaggtctat ataagcagag ctggtttagt gaaccgtcag atccgctagc gctaccggtc
661 gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
721 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc
781 acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg
841 cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac
901 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
961 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac
1021 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
1081 gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag
1141 aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag
1201 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
1261 aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac
1321 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
1381 aagtccggac tcagatctcg agctcaagct tcgaattctg cagtcgacgg taccgcgggc
1441 ccgggatcca tggactcggg ccgggacttc ctgaccctgc acggcctaca ggatgatgag
1501 gatctacagg cgctgctgaa gggcagccag ctcctgaagg tgaagtccag ctcatggagg
1561 agagagcgct tctacaagtt gcaggaggac tgcaagacca tctggcagga gtcccgcaag
1621 gtcatgcgga ccccggagtc ccagctgttc tccatcgagg acattcagga ggtgcgaatg
1681 gggcaccgca cggagggtct ggagaagttc gcccgtgatg tgcccgagga ccgctgcttc
1741 tccattgtct tcaaggacca gcgcaataca ctagacctca tcgccccatc gccagctgat
1801 gcccagcact gggtgctggg gctgcacaag atcatccacc actcaggctc catggaccag
1861 cgtcagaagc tacagcactg gattcactcc tgcttgcgaa aagctgacaa aaacaaggac
1921 aacaagatga gcttcaagga gctgcagaac ttcctgaagg agctcaacat ccagtaatct
1981 agataactga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac
2041 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
2101 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
2161 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaacgc
2221 gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
2281 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
2341 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
2401 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
2461 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
2521 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
2581 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
2641 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa
2701 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
2761 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga
2821 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
2881 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
2941 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
3001 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
3061 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
3121 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga
3181 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
3241 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
3301 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
3361 gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac
3421 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
3481 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
3541 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
3601 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
3661 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
3721 gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
3781 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
3841 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
3901 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
3961 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
4021 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
4081 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
4141 gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag
4201 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt
4261 gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc
4321 ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc
4381 ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata
4441 gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
4501 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
4561 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
4621 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
4681 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
4741 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
4801 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
4861 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
4921 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
4981 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
5041 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
5101 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
5161 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
5221 ttcctggcct tttgctggcc ttttgctcac atg
//
