| 质粒名称: | Mfn2-YFP |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 28010 |
| 存储实验室: | Richard Youle |
|
载体骨架基本信息 |
|
| 载体名称: | YFP-N1 |
| 载体出品公司: | -- |
| 载体抗性: | 卡那霉素 |
| 空载体大小: | 4700 bp |
| 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: | mitofusin 2,Mfn2 |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 2282 bp |
| 基因突变: | silent S508S |
| GenBank编号 | -- |
| 启动子 | NM_014874 |
| 标签/融合蛋白: | YFP(主干上的C终端) |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | EcoRI (未破坏) |
| 3' 克隆位点: | BamH1 (未破坏) |
| 5' 测序引物 | AGGTCTATATAAGCAGAGC |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | Mfn2-YFP was a gift from Richard Youle (Addgene plasmid # 28010 ; http://n2t.net/addgene:28010 ; RRID:Addgene_28010) |
| 参考文献部分: | Spatial and temporal association of Bax with mitochondrial fission sites, Drp1, and Mfn2 during apoptosis. Karbowski M, Lee YJ, Gaume B, Jeong SY, Frank S, Nechushtan A, Santel A, Fuller M, Smith CL, Youle RJ. J Cell Biol. 2002 Dec 23. 159(6):931-8. 10.1083/jcb.200209124 PubMed 12499352 |
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Mfn2-YFP载体 Mfn2YFP质粒基本信息
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Mfn2-YFP质粒 Mfn2YFP载体简介
Mfn2-YFP质粒序列 Mfn2YFP载体序列
LOCUS Mfn2-YFP 6984 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS Mfn2-YFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6984)
AUTHORS Karbowski M, Lee YJ, Gaume B, Jeong SY, Frank S, Nechushtan A,
Santel A, Fuller M, Smith CL, Youle RJ
TITLE Spatial and temporal association of Bax with mitochondrial fission
sites, Drp1, and Mfn2 during apoptosis.
JOURNAL J Cell Biol. 2002 Dec 23. 159(6):931-8.
PUBMED 12499352
REFERENCE 2 (bases 1 to 6984)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6984
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 68..371
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 372..575
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 526..546
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
RBS 1562..1570
/label=Shine-Dalgarno sequence
/note="full consensus sequence for ribosome-binding sites
upstream of start codons in E. coli; complementary to a
region in the 3' end of the 16S rRNA (Chen et al., 1994)"
regulatory 2931..2940
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 2937..3656
/codon_start=1
/product="enhanced YFP"
/label=EYFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(2982..3003)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(3243..3262)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 3590..3611
/label=EGFP-C
/note="EGFP, forward primer"
polyA_signal 3779..3900
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3816..3835)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3870..3889
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(3907..4362)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4044..4065)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 4256..4275
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 4389..4493
/gene="bla"
/label=AmpR promoter
promoter 4495..4852
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(4518..4538)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 4703..4838
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 4765..4784
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 4887..5681
/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(4941..4960)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 5551..5570
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(5869..5888)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 5913..5960
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 6289..6877
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6778..6797
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
61 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
121 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
181 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
241 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
301 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
361 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
421 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
481 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
541 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
601 agcgctaccg gactcagatc tcgagctcaa gcttcgaatt cgcgatgtcc ctgctcttct
661 ctcgatgcaa ctctatcgtc acagtcaaga aaaataagag acacatggct gaggtgaatg
721 catccccact taagcacttt gtcactgcca agaagaagat caatggcatt tttgagcagc
781 tgggggccta catccaggag agcgccacct tccttgaaga cacgtacagg aatgcagaac
841 tggaccccgt taccacagaa gaacaggttc tggacgtcaa aggttaccta tccaaagtga
901 gaggcatcag tgaggtgctg gctcggaggc acatgaaagt ggcttttttt ggccggacga
961 gcaatgggaa gagcaccgtg atcaatgcca tgctctggga caaagttctg ccctctggga
1021 ttggccacac caccaattgc ttcctgcggg tagagggcac agatggccat gaggcctttc
1081 tccttaccga gggctcagag gaaaagagga gtgccaagac tgtgaaccag ctggcccatg
1141 ccctccacca ggacaagcag ctccatgccg gcagcctagt gagtgtgatg tggcccaact
1201 ctaagtgccc acttctgaag gatgacctcg ttttgatgga cagccctggt attgatgtca
1261 ccacagagct ggacagctgg attgacaagt tttgtctgga tgctgatgtg tttgtgctgg
1321 tggccaactc agagtccacc ctgatgcaga cggaaaagca cttcttccac aaggtgagtg
1381 agcgtctctc ccggccaaac atcttcatcc tgaacaaccg ctgggatgca tctgcctcag
1441 agcccgagta catggaggag gtgcggcggc agcacatgga gcgttgtacc agcttcctgg
1501 tggatgagct gggcgtggtg gatcgatccc aggccgggga ccgcatcttc tttgtgtctg
1561 ctaaggaggt gctcaacgcc aggattcaga aagcccaggg catgcctgaa ggagggggcg
1621 ctctcgcaga aggctttcaa gtgaggatgt ttgagtttca gaattttgag aggagatttg
1681 aggagtgcat ctcccagtct gcagtgaaga ccaagtttga gcagcacacg gtccgggcca
1741 agcagattgc agaggcggtt cgactcatca tggactccct gcacatggcg gctcgggagc
1801 agcaggttta ctgcgaggaa atgcgtgaag agcggcaaga ccgactgaaa tttattgaca
1861 aacagctgga gctcttggct caagactata agctgcgaat taagcagatt acggaggaag
1921 tggagaggca ggtgtcgact gcaatggccg aggagatcag gcgcctctct gtactggtgg
1981 acgattacca gatggacttc cacccttctc cagtagtcct caaggtttat aagaatgagc
2041 tgcaccgcca catagaggaa ggactgggtc gaaacatgtc tgaccgctgc tccacggcca
2101 tcaccaactc cctgcagacc atgcagcagg acatgataga tggcttgaaa cccctccttc
2161 ctgtgtccgt gcggagtcag atagacatgc tggtcccacg ccagtgcttc tccctcaact
2221 atgacctaaa ctgtgacaag ctgtgtgctg acttccagga agacattgag ttccatttct
2281 ctctcggatg gaccatgctg gtgaataggt tcctgggccc caagaacagc cgtcgggcct
2341 tgatgggcta caatgaccag gtccagcgtc ccatccctct gacgccagcc aaccccagca
2401 tgcccccact gccacagggc tcgctcaccc aggaggagtt catggtttcc atggttaccg
2461 gcctggcctc cttgacatcc aggacctcca tgggcattct tgttgttgga ggagtggtgt
2521 ggaaggcagt gggctggcgg ctcattgccc tctcctttgg gctctatggc ctcctctacg
2581 tctatgagcg tctgacctgg accaccaagg ccaaggagag ggccttcaag cgccagtttg
2641 tggagcatgc cagcgagaag ctgcagcttg tcatcagcta cactggctcc aactgcagcc
2701 accaagtcca gcaggaactg tctgggacct ttgctcatct gtgtcagcaa gttgacgtca
2761 cccgggagaa cctggagcag gaaattgccg ccatgaacaa gaaaattgag gttcttgact
2821 cacttcagag caaagcaaag ctgctcagga ataaagccgg ttggttggac agtgagctca
2881 acatgttcac acaccagtac ctgcagccca gcagagcgga tccaccggtc gccaccatgg
2941 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
3001 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
3061 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
3121 tgaccacctt cggctacggc ctgcagtgct tcgcccgcta ccccgaccac atgaagcagc
3181 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
3241 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
3301 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
3361 tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca
3421 tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc
3481 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
3541 tgagctacca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
3601 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaaagcg
3661 gccgcgactc tagatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa
3721 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
3781 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
3841 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
3901 aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
3961 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
4021 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
4081 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
4141 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
4201 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
4261 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
4321 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg
4381 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
4441 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg
4501 cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
4561 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
4621 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
4681 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
4741 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
4801 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc
4861 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
4921 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
4981 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
5041 acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca
5101 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
5161 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
5221 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
5281 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
5341 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
5401 ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
5461 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
5521 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
5581 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
5641 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
5701 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
5761 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
5821 cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa
5881 ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca
5941 cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga
6001 taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca
6061 ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc
6121 catagcctca ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa
6181 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
6241 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
6301 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
6361 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
6421 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
6481 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
6541 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
6601 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
6661 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
6721 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
6781 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
6841 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
6901 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
6961 ttctgtggat aaccgtatta ccgc
//
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