| 质粒名称: | mito-PAGFP |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 23348 |
| 存储实验室: | Richard Youle |
|
载体骨架基本信息 |
|
| 载体名称: | mito-GFP (BD Biosciences) |
| 载体出品公司: | -- |
| 载体抗性: | 卡那霉素 |
| 空载体大小: | 4059 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: | PAGFP,Mitochondrial targeting sequence from subunit VIII of human cytochrome c oxidase，photoactivatable mito-GFP |
| 物种来源: | -- |
| 插入大小: | 741 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | mito（主干网上的N终端） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | BamH1(未破坏) |
| 3' 克隆位点: | Not1(未破坏) |
| 5' 测序引物 | -- |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | mito-PAGFP was a gift from Richard Youle (Addgene plasmid # 23348 ; http://n2t.net/addgene:23348 ; RRID:Addgene_23348) |
| 参考文献部分: | Quantitation of mitochondrial dynamics by photolabeling of individual organelles shows that mitochondrial fusion is blocked during the Bax activation phase of apoptosis. Karbowski M, Arnoult D, Chen H, Chan DC, Smith CL, Youle RJ. J Cell Biol. 2004 Feb 16. 164(4):493-9. 10.1083/jcb.200309082 PubMed 14769861 |
买家导航
mitoPAGFP载体基本信息
mitoPAGFP载体图谱质粒图谱和多克隆位点信息
mitoPAGFP载体简介
mitoPAGFP载体序列
LOCUS mito-PAGFP 4759 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Expresses a mitochondrial matrix-targeted photoactivable GFP that
enables detection and quantification of organelle fusion in living
cells..
ACCESSION .
VERSION .
KEYWORDS mito-PAGFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4759)
AUTHORS Karbowski M, Arnoult D, Chen H, Chan DC, Smith CL, Youle RJ
TITLE Quantitation of mitochondrial dynamics by photolabeling of
individual organelles shows that mitochondrial fusion is blocked
during the Bax activation phase of apoptosis.
JOURNAL J Cell Biol. 2004 Feb 16. 164(4):493-9.
PUBMED 14769861
REFERENCE 2 (bases 1 to 4759)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..4759
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 115..418
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 419..622
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 573..593
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
regulatory 753..762
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 759..1475
/codon_start=1
/product="photoactivatable GFP variant (Patterson et al.,
2002)"
/label=PA-GFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSHQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(804..825)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(1065..1084)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1412..1433
/label=EGFP-C
/note="EGFP, forward primer"
polyA_signal 1601..1722
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1638..1657)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 1692..1711
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(1729..2184)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(1866..1887)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2078..2097
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 2211..2315
/gene="bla"
/label=AmpR promoter
promoter 2317..2674
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(2340..2360)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 2525..2660
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2587..2606
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 2709..3503
/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(2763..2782)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3373..3392
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(3691..3710)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 3735..3782
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 4111..4699
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4600..4619
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat gcattagtta
61 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
121 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
181 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
241 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
301 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
361 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
421 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
481 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
541 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
601 ggaggtctat ataagcagag ctggtttagt gaaccgtcag atccgctagc atgtccgtcc
661 tgacgccgct gctgctgcgg ggcttgacag gctcggcccg gcggctccca gtgccgcgcg
721 ccaagatcca ttcgttgggg gatccaccgg tcgccaccat ggtgagcaag ggcgaggagc
781 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
841 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca
901 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ttcagctacg
961 gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
1021 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
1081 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
1141 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca
1201 gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggcc aacttcaaga
1261 tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc
1321 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagccac cagtccgccc
1381 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg
1441 ccgggatcac tctcggcatg gacgagctgt acaagtaaag cggccgcgac tctagatcat
1501 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
1561 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
1621 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
1681 gcattctagt tgtggtttgt ccaaactcat caatgtatct taaggcgtaa attgtaagcg
1741 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
1801 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
1861 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
1921 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
1981 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
2041 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
2101 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
2161 ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
2221 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
2281 gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg
2341 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
2401 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
2461 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
2521 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
2581 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
2641 aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat
2701 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
2761 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
2821 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
2881 atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
2941 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
3001 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
3061 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
3121 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
3181 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca
3241 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
3301 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
3361 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
3421 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
3481 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
3541 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
3601 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
3661 gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg
3721 aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt
3781 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca
3841 ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga
3901 aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc
3961 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
4021 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
4081 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
4141 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
4201 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
4261 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
4321 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
4381 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
4441 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
4501 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
4561 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
4621 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
4681 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
4741 ctggcctttt gctcacatg
//
