| 质粒名称: | GFP-P4M-SidMx2 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 51472 |
| 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 |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 新霉素（G418） |
| 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: | P4M-SidM(546-647) |
| 物种来源: | Legionella pneumophila |
| 插入大小: | 321 bp |
| GenBank编号 | DQ845395.1 |
| 启动子: | CMV |
| 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: | EcoRI（未销毁） |
| 3’克隆位点: | SalI（未被破坏） |
| 5’测序引物 | EGFP-C |
| 3’测序引物 | SV40-R |
|
文献引用方法 |
|
| 材料和方法部分: |
GFP-P4M-SidMx2 was a gift from Tamas Balla (Addgene plasmid # 51472 ; http://n2t.net/addgene:51472 ; RRID:Addgene_51472)
|
| 参考文献部分: |
A novel probe for phosphatidylinositol 4-phosphate reveals multiple pools beyond the Golgi. Hammond GR, Machner MP, Balla T. J Cell Biol. 2014 Apr 7. 10.1083/jcb.201312072 PubMed 24711504
|
买家导航
GFP-P4M-SidMx2载体质粒基本信息
GFP-P4M-SidMx2质粒图谱载体图谱和GFP-P4M-SidMx2载体序列质粒序列多克隆位点信息
GFP-P4M-SidMx2质粒载体简介
GFP-P4M-SidMx2质粒序列载体序列
LOCUS GFP-P4M-SidMx2 5394 bp DNA circular SYN 15-DEC-2025
DEFINITION Biosensor for PtdIns4P .
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5394)
AUTHORS Hammond GR, Machner MP, Balla T
TITLE A novel probe for phosphatidylinositol 4-phosphate reveals multiple
pools beyond the Golgi.
JOURNAL J Cell Biol. 2014 Apr 7.
PUBMED 24711504
REFERENCE 2 (bases 1 to 5394)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Dec 15, 2025 from SnapGene Server 8.0.1
https://www.biofeng.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "J Cell
Biol. 2014 Apr 7."
COMMENT Sequence Label: GFP-P4M-SidMx2
FEATURES Location/Qualifiers
source 1..5394
/mol_type="other DNA"
/organism="synthetic DNA construct"
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 607..616
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS join(613..615,616..618,619..1329)
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
CDS join(613..615,616..618,619..1329)
/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(658..679)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(919..938)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1266..1287
/label=EGFP-C
/note="EGFP, forward primer"
CDS 1393..1698
/codon_start=1
/product="high-affinity phosphatidylinositol 4-phosphate
binding module of SidM (DrrA) from Legionella pneumophila
(Hammond et al., 2014)"
/label=P4M
/translation="TASTENFKNVKEKYQQMRGDALKTEILADFKDKLAEATDEQSLKQ
IVAELKSKDEYRILAKGQGLTTQLLGLKTSSVSSFEKMVEETRESIKSQERQTIKIK"
CDS 1744..2049
/codon_start=1
/product="high-affinity phosphatidylinositol 4-phosphate
binding module of SidM (DrrA) from Legionella pneumophila
(Hammond et al., 2014)"
/label=P4M
/translation="TASTENFKNVKEKYQQMRGDALKTEILADFKDKLAEATDEQSLKQ
IVAELKSKDEYRILAKGQGLTTQLLGLKTSSVSSFEKMVEETRESIKSQERQTIKIK"
polyA_signal 2182..2303
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2219..2238)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 2273..2292
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(2310..2765)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2447..2468)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2659..2678
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 2792..2896
/gene="bla"
/label=AmpR promoter
promoter 2898..3255
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
enhancer 2898..3089
/label=SV40 enhancer
/note="enhancer for the SV40 early promoter (Herr, 1993)"
primer_bind complement(2921..2941)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 3106..3241
/label=SV40 ori
/note="SV40 origin of replication"
CDS 3290..4084
/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 3290..4084
/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(3344..3363)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3954..3973
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(4272..4291)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 4316..4363
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 4692..5280
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5181..5200
/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 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
1321 ctgtacaagt ccggactcag atctcgagct caagcttcga attctggtgg tagtgcttcc
1381 ggactcagat ccacggcaag cacggaaaac tttaaaaatg ttaaagaaaa atatcagcaa
1441 atgcgaggtg atgctttaaa aacagaaatc ctggctgatt tcaaggataa actggctgaa
1501 gctaccgatg aacagagcct taagcaaatt gttgctgaat tgaaaagtaa agatgaatat
1561 agaattttgg ctaaggggca agggttaaca acccagcttt tagggttaaa gacaagttca
1621 gtgtcttcat ttgagaaaat ggttgaggaa acaagagaaa gtattaagtc tcaagaaaga
1681 caaaccatta agataaaagg tggtagtgct gtcgacggtg gtagtgcttc cggactcaga
1741 tccacggcaa gcacggaaaa ctttaaaaat gttaaagaaa aatatcagca aatgcgaggt
1801 gatgctttaa aaacagaaat cctggctgat ttcaaggata aactggctga agctaccgat
1861 gaacagagcc ttaagcaaat tgttgctgaa ttgaaaagta aagatgaata tagaattttg
1921 gctaaggggc aagggttaac aacccagctt ttagggttaa agacaagttc agtgtcttca
1981 tttgagaaaa tggttgagga aacaagagaa agtattaagt ctcaagaaag acaaaccatt
2041 aagataaaat aaggatccac cggatctaga taactgatca taatcagcca taccacattt
2101 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa
2161 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc
2221 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
2281 tccaaactca tcaatgtatc ttaacgcgta aattgtaagc gttaatattt tgttaaaatt
2341 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
2401 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
2461 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
2521 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
2581 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
2641 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
2701 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
2761 cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
2821 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
2881 ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg
2941 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
3001 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
3061 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
3121 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
3181 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
3241 ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca
3301 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
3361 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
3421 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc
3481 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
3541 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
3601 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
3661 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
3721 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
3781 gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct
3841 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
3901 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
3961 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
4021 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
4081 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga
4141 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
4201 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg
4261 gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca
4321 ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg
4381 tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc
4441 gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc
4501 aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat
4561 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
4621 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
4681 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
4741 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
4801 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
4861 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
4921 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
4981 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
5041 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
5101 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
5161 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
5221 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
5281 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
5341 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat gcat
//
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