| 质粒名称: | FAB(ER-mito) |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 214427 |
| 存储实验室: | Chi-Lun Chang |
|
载体骨架基本信息 |
|
| 载体名称: | RpsAN-mCh-Sec61_IRES_CFAST10-20nm-Halo-mito |
| 载体出品公司: | Clontech |
| 完整质粒大小: | 7508bp |
| 载体类型: | 哺乳细胞表达 |
| 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: | FRB-V5 |
| 物种来源: | H. sapiens (human) |
| 启动子: | CMV |
| 标签/融合蛋白: |
RspAN-FRB-V5 (N terminal on insert)
CFAST10-Halo (C terminal on insert)
|
| 克隆方法: | Gibson Cloning |
|
文献引用方法 |
|
| 材料和方法部分: | FAB(ER-mito) was a gift from Chi-Lun Chang (Addgene plasmid # 214427 ; http://n2t.net/addgene:214427 ; RRID:Addgene_214427) |
| 参考文献部分: | A fluorogenic complementation tool kit for interrogating lipid droplet-organelle interaction. Li X, Gamuyao R, Wu ML, Cho WJ, King SV, Petersen RA, Stabley DR, Lindow C, Climer LK, Shirinifard A, Ferrara F, Throm RE, Robinson CG, Zhou Y, Carisey AF, Tebo AG, Chang CL. J Cell Biol. 2024 Sep 2;223(9):e202311126. doi: 10.1083/jcb.202311126. Epub 2024 Jul 1. 10.1083/jcb.202311126 PubMed 38949658 |
买家导航
FAB(ER-mito)载体质粒基本信息
FAB(ER-mito)质粒图谱载体图谱和FAB(ER-mito)载体序列质粒序列多克隆位点信息
质粒图谱.png)
FAB(ER-mito)质粒载体简介
FAB(ER-mito)质粒序列载体序列
LOCUS FAB(ER-mito) 7508 bp DNA circular SYN 14-APR-2025
DEFINITION Detection of ER-mito contact site.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7508)
AUTHORS Li X, Gamuyao R, Wu ML, Cho WJ, King SV, Petersen RA, Stabley DR,
Lindow C, Climer LK, Shirinifard A, Ferrara F, Throm RE, Robinson
CG, Zhou Y, Carisey AF, Tebo AG, Chang CL
TITLE A fluorogenic complementation tool kit for interrogating lipid
droplet-organelle interaction.
JOURNAL J Cell Biol. 2024 Sep 2;223(9):e202311126. doi:
10.1083/jcb.202311126. Epub 2024 Jul 1.
PUBMED 38949658
REFERENCE 2 (bases 1 to 7508)
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."; date: "2024-09-2"; volume: "223(9):e202311126. doi"; pages:
" 10.1083/jcb.202311126. Epub 2024 Jul 1"
COMMENT Sequence Label: FAB(ER-mito)
FEATURES Location/Qualifiers
source 1..7508
/mol_type="other DNA"
/organism="synthetic DNA construct"
regulatory 154..163
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
misc_feature join(190..198,199..201,202..776)
/label=IRES2
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
primer_bind complement(363..382)
/label=IRES-F
/note="IRES internal ribosome entry site, forward primer"
primer_bind 592..609
/label=IRES reverse
/note="IRES internal ribosome entry site, reverse primer.
Also called pCDH-rev"
primer_bind complement(846..865)
/label=pMT2-F
/note="Synthetic intron, forward primer"
intron 848..1077
/label=chimeric intron
/note="chimera between introns from adenovirus and
immunoglobulin heavy chain genes"
CDS complement(1182..1472)
/codon_start=1
/gene="SEC61B"
/product="human Sec61-beta subunit of the translocon in the
endoplasmic reticulum"
/label=Sec61-beta
/translation="MPGPTPSGTNVGSSGRSPSKAVAARAAGSTVRQRKNASCGTRSAG
RTTSAGTGGMWRFYTEDSPGLKVGPVPVLVMSLLFIASVFMLHIWGKYTRS"
CDS complement(1518..1559)
/codon_start=1
/product="epitope tag from simian virus 5"
/label=V5 tag
/translation="GKPIPNPLLGLDST"
regulatory 1898..1907
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
regulatory 2288..2297
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
promoter complement(2324..2527)
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind complement(2353..2373)
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
enhancer 2528..2831
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
rep_origin complement(3006..3594)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3086..3105)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
polyA_signal 3923..3970
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
primer_bind 3995..4014
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
CDS complement(4202..4996)
/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 complement(4202..4996)
/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(4313..4332)
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind 4923..4942
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
promoter complement(5031..5388)
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 5045..5180
/label=SV40 ori
/note="SV40 origin of replication"
enhancer 5197..5388
/label=SV40 enhancer
/note="enhancer for the SV40 early promoter (Herr, 1993)"
primer_bind 5345..5365
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter complement(5390..5494)
/gene="bla"
/label=AmpR promoter
rep_origin 5521..5976
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(5608..5627)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 5818..5839
/label=F1ori-F
/note="F1 origin, forward primer"
polyA_signal 5983..6104
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(5994..6013)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 6048..6067
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
CDS complement(6457..7347)
/codon_start=1
/product="modified bacterial dehalogenase that forms
covalent bonds with chloroalkane derivatives"
/label=HaloTag(R)
/translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
IGSEIARWLSTLEISG"
regulatory 7344..7353
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
ORIGIN
1 cggcaccttc tagaattcta cctccggcac ttccaccagc actgcctcca gctgagaccg
61 gtagggtgga acctccagaa ccgccgctgc ctccggagcc gcctccggtt tcggtttcac
121 tagtccgttt cacaaagacc caatagctgt cacccatggt ggctagcgga tctgacggcg
181 cgcctcccgg gttgtggcaa gcttatcatc gtgtttttca aaggaaaacc acgtccccgt
241 ggttcggggg gcctagacgt ttttttaacc tcgactaaac acatgtaaag catgtgcacc
301 gaggccccag atcagatccc atacaatggg gtaccttctg ggcatccttc agccccttgt
361 tgaatacgct tgaggagagc catttgactc tttccacaac tatccaactc acaacgtggc
421 actggggttg tgccgccttt gcaggtgtat cttatacacg tggcttttgg ccgcagaggc
481 acctgtcgcc aggtgggggg ttccgctgcc tgcaaagggt cgctacagac gttgtttgtc
541 ttcaagaagc ttccagagga actgcttcct tcacgacatt caacagacct tgcattcctt
601 tggcgagagg ggaaagaccc ctaggaatgc tcgtcaagaa gacagggcca ggtttccggg
661 ccctcacatt gccaaaagac ggcaatatgg tggaaaataa catatagaca aacgcacacc
721 ggccttattc caagcggctt cggccagtaa cgttaggggg gggggaggga gagggggggg
781 gggagagggg cggaattggc cgccctagat gcatgctcga cctgcagttg gacctgggag
841 tggacacctg tggagagaaa ggcaaagtgg atgtcattgt cactcaagtg tatggccaga
901 tctcaagcct gccacacctc aagcttgaca acaaaaagat tgtcttttct gaccagatgg
961 acgcggccac cctcaaaggc atcaccgcgg gccaggtgaa tatcaaatcc tcctcgtttt
1021 tggaaactga caatcttagc gcagaagtca tgcccgcttt tgagagggag tactcacccc
1081 aacagctggc cctcgcagac agcgaattaa ttccagcaca ctggatcagt tatctatgcg
1141 gccgcttagt cctcggccat gtaccgtcga ctgcagaatt cctacgaacg agtgtacttg
1201 ccccaaatgt gcaacataaa tacagaagcg atgaacagaa gactcataac caatactgga
1261 acagggccaa ctttgagccc aggtgaatct tctgtgtaga atcgccacat ccccccggtg
1321 cctgccgagg ttgtgcggcc tgcactcctt gtcccacagc tggcattttt cctctgccgg
1381 acagtggatc ccgccgcccg ggcggccact gctttgctgg gagagcgccc tgaggatccc
1441 acgttagtgc cactgggggt cggaccaggc atagatctga gtccggactt gtacagctcg
1501 tccatgccat taagtttggt gctgtccagg cccagcaggg ggttggggat gggcttgcca
1561 ccgctcccac tacctccgct gccacttgag attcgtcgga acacatgata atagaggtcc
1621 caggcttggg tgaggtcctt gacattccct gatttcatgt acttcctgca ccactcttgg
1681 gcctccatta aatctcgacc ataggcctga ttaaaggatg tttccttcag agtctggggg
1741 ccccgttcca tcatagcatg caagggctcc agcacctcaa acatgccttt cacgttcctt
1801 tccccaaagt acaaacgaga tgcctcttcc aggccttcat gccacatctc gccccctcct
1861 gatccccctc cgccgctgcc ctcctcgccc ttgctcacca tggtggcgac cggtagagat
1921 ccccctccgc cgctgccgcc tcctccggag gtcatggcct tcttcatgtg caccttcacc
1981 ttggtggggc ctctgctggt ggggatcatc cactcgaaca tggtgttcag gtcgcccttc
2041 tgcacgccct ccttgaatct gccctggaac tccttgctct gggtgccggg ggccacgtcg
2101 gtgaagaagt tcttgccgat cacggtcttg gggtctctgc cggtgatggt gccctcggcg
2161 gcgttgtagt ggatgatctt gccgttgccg tccagctgga tggcgccgaa ggccagcttg
2221 tccagggcct tgtcgtccat cttggccagg ctgttctcga tgtcgtcgcc gccgaatctc
2281 acggtctcca tggtggctag cggatctgac ggttcactaa accagctctg cttatataga
2341 cctcccaccg tacacgccta ccgcccattt gcgtcaatgg ggcggagttg ttacgacatt
2401 ttggaaagtc ccgttgattt tggtgccaaa acaaactccc attgacgtca atggggtgga
2461 gacttggaaa tccccgtgag tcaaaccgct atccacgccc attgatgtac tgccaaaacc
2521 gcatcaccat ggtaatagcg atgactaata cgtagatgta ctgccaagta ggaaagtccc
2581 ataaggtcat gtactgggca taatgccagg cgggccattt accgtcattg acgtcaatag
2641 ggggcgtact tggcatatga tacacttgat gtactgccaa gtgggcagtt taccgtaaat
2701 actccaccca ttgacgtcaa tggaaagtcc ctattggcgt tactatggga acatacgtca
2761 ttattgacgt caatgggcgg gggtcgttgg gcggtcagcc aggcgggcca tttaccgtaa
2821 gttatgtaac gcggaactcc atatatgggc tatgaactaa tgaccccgta attgattact
2881 attaataact aatgcatggc ggtaatacgg ttatccacag aatcagggga taacgcagga
2941 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
3001 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
3061 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
3121 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
3181 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
3241 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
3301 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
3361 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
3421 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
3481 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
3541 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
3601 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
3661 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
3721 tttaaatcaa tctaaagtat atatgagtaa cctgaggcta tggcagggcc tgccgccccg
3781 acgttggctg cgagccctgg gccttcaccc gaacttgggg ggtggggtgg ggaaaaggaa
3841 gaaacgcggg cgtattggcc ccaatggggt ctcggtgggg tatcgacaga gtgccagccc
3901 tgggaccgaa ccccgcgttt atgaacaaac gacccaacac cgtgcgtttt attctgtctt
3961 tttattgccg tcatagcgcg ggttccttcc ggtattgtct ccttccgtgt ttcagttagc
4021 ctccccctag ggtgggcgaa gaactccagc atgagatccc cgcgctggag gatcatccag
4081 ccggcgtccc ggaaaacgat tccgaagccc aacctttcat agaaggcggc ggtggaatcg
4141 aaatctcgtg atggcaggtt gggcgtcgct tggtcggtca tttcgaaccc cagagtcccg
4201 ctcagaagaa ctcgtcaaga aggcgataga aggcgatgcg ctgcgaatcg ggagcggcga
4261 taccgtaaag cacgaggaag cggtcagccc attcgccgcc aagctcttca gcaatatcac
4321 gggtagccaa cgctatgtcc tgatagcggt ccgccacacc cagccggcca cagtcgatga
4381 atccagaaaa gcggccattt tccaccatga tattcggcaa gcaggcatcg ccatgggtca
4441 cgacgagatc ctcgccgtcg ggcatgctcg ccttgagcct ggcgaacagt tcggctggcg
4501 cgagcccctg atgctcttcg tccagatcat cctgatcgac aagaccggct tccatccgag
4561 tacgtgctcg ctcgatgcga tgtttcgctt ggtggtcgaa tgggcaggta gccggatcaa
4621 gcgtatgcag ccgccgcatt gcatcagcca tgatggatac tttctcggca ggagcaaggt
4681 gagatgacag gagatcctgc cccggcactt cgcccaatag cagccagtcc cttcccgctt
4741 cagtgacaac gtcgagcaca gctgcgcaag gaacgcccgt cgtggccagc cacgatagcc
4801 gcgctgcctc gtcttgcagt tcattcaggg caccggacag gtcggtcttg acaaaaagaa
4861 ccgggcgccc ctgcgctgac agccggaaca cggcggcatc agagcagccg attgtctgtt
4921 gtgcccagtc atagccgaat agcctctcca cccaagcggc cggagaacct gcgtgcaatc
4981 catcttgttc aatcatgcga aacgatcctc atcctgtctc ttgatcgatc tttgcaaaag
5041 cctaggcctc caaaaaagcc tcctcactac ttctggaata gctcagaggc cgaggcggcc
5101 tcggcctctg cataaataaa aaaaattagt cagccatggg gcggagaatg ggcggaactg
5161 ggcggagtta ggggcgggat gggcggagtt aggggcggga ctatggttgc tgactaattg
5221 agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc acacctggtt
5281 gctgactaat tgagatgcat gctttgcata cttctgcctg ctggggagcc tggggacttt
5341 ccacacccta actgacacac attccacagc tggttctttc cgcctcagga ctcttccttt
5401 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
5461 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
5521 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
5581 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
5641 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
5701 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
5761 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
5821 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
5881 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
5941 acgcgaattt taacaaaata ttaacgctta caatttacgc gttaagatac attgatgagt
6001 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
6061 ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca
6121 ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc
6181 tctacaaatg tggtatggct gattatgatc agttatctag agttaattaa ttggccctcg
6241 agttacttgt accgatccgg tggatcctta tcaaagttgt tgccggtatc tcatgaaagc
6301 ccaggctgct accatggtga gggcaaacac tggcaccagc accataaata tgggaatacc
6361 acttgggtcc ccttcacctc gatgtcctat aggtccattc tgcacctgca aagaattcga
6421 agcttgagct cgagatctga gtccggactt gtacatgccg gaaatctcga gcgtcgacag
6481 ccagcgcgcg atctcgctgc cgatcaggtc cgggttgtct tcttgcagca gattcagacc
6541 cgggccgatg tccacagcct tgcagttagg caggcttttg gccaggcgag cggcttcggc
6601 cggtgggatc agaacgcctg gggtgcccca gaacagcagc ttcgggacag gggactggtg
6661 cagccagtcc atgtattctt cgaccagcgc gacgatgttc gctggctcac cggcgattgg
6721 cagctcgttt gggaagcgcc acagtggctc gcggtcaaca ggattcagga acggctcgcg
6781 gtaatggtcc atctcgactt cagtcagcgg gcggacgaca cccatcggca gcgtaccctc
6841 gataaaaacg ttctgatcga tgatcagctt gcggccgacg tcggtggtgc ggaaggcctg
6901 gaaggtctcg cgggcaaatt ctggccattc gtcccaggtc gggatagggc ggatgaactc
6961 cataaatgca atacctttga cgcgctctgg attgcgcttg gcccagtgga aacccagagc
7021 ggagccccag tcgtgaatga ccaggacgac ctcttccaga cccagggctt cgatgaaggc
7081 atccatgaag cggacgtggt cgtcgaagaa ataacccagg tctggtttgt cggatttgcc
7141 cataccgatc aggtctggag caatgcagcg atgggtcggt gcaacatgcg ggatgatgtt
7201 gcgccacacg taggaggagg tcgggttacc gtgcaggaac agcacagggg tgccatcgcg
7261 cggaccaaca tcgacgtagt gcatgcgctc gcccaggact tccacataat gggggtcgaa
7321 tggaaagcca gtaccgattt ctgccatggt ggcgaccggt gtcgaacctc ccccacccga
7381 tccgccacct ccagatccac ctccgcctga accgccacca cctgatgctc tagctgcagc
7441 ctccctggct gcagcttctc tagcagctgc ctcacgtgca gctgcttccc tagcggcagc
7501 ctctcttg
//
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FAB(ER-mito)质粒载体应用举例
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