| 质粒名称: | NES-mCherry-P4Mx1 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 108143 |
| 存储实验室: | Gerry Hammond |
|
载体骨架基本信息 |
|
| 载体名称: | pmCherry-C1 |
| 载体出品公司: | -- |
| 空载体大小: | -- |
| 完整质粒大小: | 5067 bp |
| 载体修饰: | -- |
| 载体类型: | 哺乳动物表达 |
| 筛选标记: | -- |
| 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: | X.leavis map2k1.L(32-44):mCherry:L. penomophila SidM(546-647) |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子: | CMV |
| 标签/融合蛋白: | -- |
| 克隆方法: | Gibson Cloning |
| 5’克隆位点: | -- |
| 3’克隆位点: | -- |
| 5’测序引物 | -- |
| 3’测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | NES-mCherry-P4Mx1 was a gift from Gerry Hammond (Addgene plasmid # 108143 ; http://n2t.net/addgene:108143 ; RRID:Addgene_108143) |
| 参考文献部分: | SAC1 degrades its lipid substrate PtdIns4P in the endoplasmic reticulum to maintain a steep chemical gradient with donor membranes. Zewe JP, Wills RC, Sangappa S, Goulden BD, Hammond GR. Elife. 2018 Feb 20;7. pii: 35588. doi: 10.7554/eLife.35588. 35588 [pii] PubMed 29461204 |
买家导航
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 NES-mCherry-P4Mx1 5067 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION PI4P biosensor .
ACCESSION .
VERSION .
KEYWORDS NES-mCherry-P4Mx1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5067)
AUTHORS Zewe JP, Wills RC, Sangappa S, Goulden BD, Hammond GR
TITLE SAC1 degrades its lipid substrate PtdIns4P in the endoplasmic
reticulum to maintain a steep chemical gradient with donor
membranes.
JOURNAL Elife. 2018 Feb 20;7. pii: 35588. doi: 10.7554/eLife.35588.
PUBMED 29461204
REFERENCE 2 (bases 1 to 5067)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..5067
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 49..352
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 353..556
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 507..527
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
regulatory 583..592
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
regulatory 640..649
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 646..1353
/codon_start=1
/product="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
/label=mCherry
/note="mammalian codon-optimized"
/translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
EGRHSTGGMDELYK"
primer_bind complement(791..809)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind 1059..1078
/label=mCherry-F
/note="mCherry, forward primer"
CDS 1369..1674
/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 1843..1964
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1880..1899)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 1934..1953
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(1971..2426)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2108..2129)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2320..2339
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 2453..2557
/gene="bla"
/label=AmpR promoter
promoter 2559..2916
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(2582..2602)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 2767..2902
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2829..2848
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 2951..3745
/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(3005..3024)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3615..3634
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(3933..3952)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 3977..4024
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 4353..4941
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4842..4861
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
61 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
121 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
181 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
241 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
301 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
361 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
421 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
481 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
541 ctatataagc agagctggtt tagtgaaccg tcagatccgc tagccaccat ggctctgcag
601 aaaaagttgg aagagcttga gctggatgag gcaccggtcg ccaccatggt gagcaagggc
661 gaggaggata acatggccat catcaaggag ttcatgcgct tcaaggtgca catggagggc
721 tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc
781 acccagaccg ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc
841 ctgtcccctc agttcatgta cggctccaag gcctacgtga agcaccccgc cgacatcccc
901 gactacttga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag
961 gacggcggcg tggtgaccgt gacccaggac tcctccctgc aggacggcga gttcatctac
1021 aaggtgaagc tgcgcggcac caacttcccc tccgacggcc ccgtaatgca gaagaagacc
1081 atgggctggg aggcctcctc cgagcggatg taccccgagg acggcgccct gaagggcgag
1141 atcaagcaga ggctgaagct gaaggacggc ggccactacg acgctgaggt caagaccacc
1201 tacaaggcca agaagcccgt gcagctgccc ggcgcctaca acgtcaacat caagttggac
1261 atcacctccc acaacgagga ctacaccatc gtggaacagt acgaacgcgc cgagggccgc
1321 cactccaccg gcggcatgga cgagctgtac aagtccggac tcagatccac ggcaagcacg
1381 gaaaacttta aaaatgttaa agaaaaatat cagcaaatgc gaggtgatgc tttaaaaaca
1441 gaaatcctgg ctgatttcaa ggataaactg gctgaagcta ccgatgaaca gagccttaag
1501 caaattgttg ctgaattgaa aagtaaagat gaatatagaa ttttggctaa ggggcaaggg
1561 ttaacaaccc agcttttagg gttaaagaca agttcagtgt cttcatttga gaaaatggtt
1621 gaggaaacaa gagaaagtat taagtctcaa gaaagacaaa ccattaagat aaaataagaa
1681 ttcccgggtc gactcgacgg taccgcgggc ccgggatcca ccggatctag ataactgatc
1741 ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc
1801 cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct
1861 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
1921 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaacgcgt aaattgtaag
1981 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
2041 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag
2101 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
2161 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt
2221 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
2281 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
2341 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
2401 gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac
2461 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
2521 ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa gaaccagctg
2581 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
2641 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
2701 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
2761 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
2821 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
2881 tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga caggatgagg
2941 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
3001 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
3061 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
3121 gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
3181 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
3241 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
3301 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
3361 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
3421 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag
3481 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
3541 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
3601 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
3661 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
3721 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
3781 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
3841 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
3901 gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac aataccggaa
3961 ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt
4021 gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc
4081 cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt
4141 gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc ctcaggttac
4201 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
4261 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
4321 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
4381 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
4441 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
4501 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
4561 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
4621 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
4681 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
4741 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
4801 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
4861 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
4921 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
4981 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
5041 attaccgcca tgcattagtt attaata
//
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