| 质粒名称: | mCherry-P4M-SidM |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 51471 |
| 存储实验室: | Tamas Balla |
|
载体骨架基本信息 |
|
| 载体名称: | pmCherry-N1 |
| 载体出品公司: | Clontech |
| 载体抗性: | 卡那霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 新霉素（G418） |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
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 |
| 基因突变: | -- |
| GenBank编号 | DQ845395.1 |
| 启动子 | CMV |
| 标签/融合蛋白: | mCherry（插入件上的N端子） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | AccIII(未破坏) |
| 3' 克隆位点: | EcoRI (未破坏) |
| 5' 测序引物 | CMV-F |
| 3' 测序引物 | SV40-R |
|
文献引用方法 |
|
| 材料和方法部分: | mCherry-P4M-SidM was a gift from Tamas Balla (Addgene plasmid # 51471 ; http://n2t.net/addgene:51471 ; RRID:Addgene_51471) |
| 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
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 mCherry-P4M-SidM 5022 bp ds-DNA circular SYN 12-NOV-2018
DEFINITION Biosensor for PtdIns4P.
ACCESSION .
VERSION .
KEYWORDS mCherry-P4M-SidM
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5022)
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 5022)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Nov 12, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5022
/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 607..616
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 613..1320
/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(758..776)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind 1026..1045
/label=mCherry-F
/note="mCherry, forward primer"
polyA_signal 1810..1931
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1847..1866)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 1901..1920
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(1938..2393)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2075..2096)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2287..2306
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 2420..2524
/gene="bla"
/label=AmpR promoter
promoter 2526..2883
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(2549..2569)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 2734..2869
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2796..2815
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 2918..3712
/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(2972..2991)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3582..3601
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(3900..3919)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 3944..3991
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 4320..4908
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4809..4828
/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 gaggataaca tggccatcat caaggagttc
661 atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc
721 gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt
781 ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc
841 tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc
901 aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc
961 tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc
1021 gacggccccg taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac
1081 cccgaggacg gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc
1141 cactacgacg ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc
1201 gcctacaacg tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg
1261 gaacagtacg aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag
1321 tccggactca gatccacggc aagcacggaa aactttaaaa atgttaaaga aaaatatcag
1381 caaatgcgag gtgatgcttt aaaaacagaa atcctggctg atttcaagga taaactggct
1441 gaagctaccg atgaacagag ccttaagcaa attgttgctg aattgaaaag taaagatgaa
1501 tatagaattt tggctaaggg gcaagggtta acaacccagc ttttagggtt aaagacaagt
1561 tcagtgtctt catttgagaa aatggttgag gaaacaagag aaagtattaa gtctcaagaa
1621 agacaaacca ttaagataaa ataagaattc ccgggtcgac tcgacggtac cgcgggcccg
1681 ggatccaccg gatctagata actgatcata atcagccata ccacatttgt agaggtttta
1741 cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt
1801 gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
1861 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
1921 aatgtatctt aacgcgtaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt
1981 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
2041 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
2101 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
2161 acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
2221 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
2281 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac
2341 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg
2401 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
2461 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
2521 gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc
2581 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
2641 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
2701 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
2761 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
2821 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
2881 aaagatcgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
2941 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
3001 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
3061 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc
3121 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
3181 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
3241 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
3301 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
3361 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
3421 cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg tcgtgaccca
3481 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
3541 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
3601 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
3661 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
3721 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
3781 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
3841 atcctccagc gcggggatct catgctggag ttcttcgccc accctagggg gaggctaact
3901 gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag
3961 aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg
4021 cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt
4081 tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg
4141 gcaggccctg ccatagcctc aggttactca tatatacttt agattgattt aaaacttcat
4201 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
4261 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
4321 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
4381 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
4441 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
4501 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
4561 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
4621 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
4681 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
4741 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
4801 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
4861 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
4921 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
4981 ttatcccctg attctgtgga taaccgtatt accgccatgc at
//
