| 质粒名称: | GFP-P4M-SidM |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 51469 |
| 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: | 4038 bp |
| 完整质粒大小: | 5052bp |
| 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: | 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: | 321bp |
| GenBank编号 | DQ845395.1 |
| 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: | SalI (not destroyed) |
| 3’克隆位点: | BamHI (not destroyed) |
| 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-SidM was a gift from Tamas Balla (Addgene plasmid # 51469 ; http://n2t.net/addgene:51469 ; RRID:Addgene_51469) |
| 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-SidM载体质粒基本信息
GFP-P4M-SidM质粒图谱载体图谱和GFP-P4M-SidM载体序列质粒序列多克隆位点信息
GFP-P4M-SidM质粒载体简介
GFP-P4M-SidM质粒序列载体序列
LOCUS pEGFP-RhoA Biosensor 5052 bp DNA circular SYN 14-APR-2025
DEFINITION Biosensor for PtdIns4P.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5052)
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 5052)
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: "J Cell
Biol. 2014 Apr 7."
COMMENT Sequence Label: GFP-P4M-SidM
FEATURES Location/Qualifiers
source 1..5052
/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 1402..1707
/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 1840..1961
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1877..1896)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 1931..1950
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(1968..2423)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2105..2126)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2317..2336
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 2450..2554
/gene="bla"
/label=AmpR promoter
promoter 2556..2913
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
enhancer 2556..2747
/label=SV40 enhancer
/note="enhancer for the SV40 early promoter (Herr, 1993)"
primer_bind complement(2579..2599)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 2764..2899
/label=SV40 ori
/note="SV40 origin of replication"
CDS 2948..3742
/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 2948..3742
/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(3002..3021)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3612..3631
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(3930..3949)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 3974..4021
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 4350..4938
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4839..4858
/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 attctgcagt cgacggtggt
1381 agtgcttccg gactcagatc cacggcaagc acggaaaact ttaaaaatgt taaagaaaaa
1441 tatcagcaaa tgcgaggtga tgctttaaaa acagaaatcc tggctgattt caaggataaa
1501 ctggctgaag ctaccgatga acagagcctt aagcaaattg ttgctgaatt gaaaagtaaa
1561 gatgaatata gaattttggc taaggggcaa gggttaacaa cccagctttt agggttaaag
1621 acaagttcag tgtcttcatt tgagaaaatg gttgaggaaa caagagaaag tattaagtct
1681 caagaaagac aaaccattaa gataaaataa ggatccaccg gatctagata actgatcata
1741 atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc
1801 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat
1861 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
1921 cattctagtt gtggtttgtc caaactcatc aatgtatctt aacgcgtaaa ttgtaagcgt
1981 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
2041 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
2101 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
2161 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
2221 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
2281 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
2341 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
2401 taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
2461 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
2521 ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg
2581 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
2641 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
2701 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
2761 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
2821 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
2881 ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc
2941 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
3001 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
3061 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
3121 tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
3181 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
3241 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
3301 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
3361 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
3421 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat
3481 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
3541 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
3601 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
3661 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
3721 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
3781 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
3841 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
3901 ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat accggaagga
3961 acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt
4021 cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat
4081 tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa
4141 ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca
4201 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
4261 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
4321 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
4381 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
4441 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt
4501 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
4561 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
4621 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
4681 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
4741 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
4801 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
4861 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
4921 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
4981 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
5041 accgccatgc at
//
