| 质粒名称: | mCherry-Parkin |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 23956 |
| 存储实验室: | Richard Youle |
|
载体骨架基本信息 |
|
| 载体名称: | mCherry-C1 |
| 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: | Park2,Parkin |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 1300 bp |
| 基因突变: | -- |
| GenBank编号 | NM_004562 |
| 启动子 | -- |
| 标签/融合蛋白: | mCherry（主干上的N终端） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | HindIII(未破坏) |
| 3' 克隆位点: | BamH1(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: | mCherry-Parkin was a gift from Richard Youle (Addgene plasmid # 23956 ; http://n2t.net/addgene:23956 ; RRID:Addgene_23956) |
| 参考文献部分: | Parkin is recruited selectively to impaired mitochondria and promotes their autophagy. Narendra D, Tanaka A, Suen DF, Youle RJ. J Cell Biol. 2008 Dec 1. 183(5):795-803. 10.1083/jcb.200809125 PubMed 19029340 |
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mCherryParkin载体基本信息
mCherryParkin载体图谱质粒图谱和多克隆位点信息
mCherryParkin载体简介
mCherryParkin载体序列
LOCUS mCherry-Parkin 6087 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Mammalian expression of human Park2 fused to mCherry.
ACCESSION .
VERSION .
KEYWORDS mCherry-Parkin
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6087)
AUTHORS Narendra D, Tanaka A, Suen DF, Youle RJ
TITLE Parkin is recruited selectively to impaired mitochondria and
promotes their autophagy.
JOURNAL J Cell Biol. 2008 Dec 1. 183(5):795-803.
PUBMED 19029340
REFERENCE 2 (bases 1 to 6087)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6087
/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 661..670
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 667..1374
/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(812..830)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind 1080..1099
/label=mCherry-F
/note="mCherry, forward primer"
polyA_signal 2929..3050
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2966..2985)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3020..3039
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(3057..3512)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3194..3215)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 3406..3425
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 3539..3643
/gene="bla"
/label=AmpR promoter
promoter 3645..4002
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(3668..3688)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 3853..3988
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3915..3934
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 4037..4831
/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(4091..4110)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 4701..4720
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(5019..5038)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 5063..5110
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 5439..6027
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5928..5947
/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 gctaccggtc
661 gccaccatgg tgagcaaggg cgaggaggat aacatggcca tcatcaagga gttcatgcgc
721 ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc
781 gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc
841 ctgcccttcg cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg
901 aagcaccccg ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg
961 gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg
1021 caggacggcg agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc
1081 cccgtaatgc agaagaagac catgggctgg gaggcctcct ccgagcggat gtaccccgag
1141 gacggcgccc tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac
1201 gacgctgagg tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac
1261 aacgtcaaca tcaagttgga catcacctcc cacaacgagg actacaccat cgtggaacag
1321 tacgaacgcg ccgagggccg ccactccacc ggcggcatgg acgagctgta caagtccgga
1381 ctcagatctc gagctcaagc tttgatagtg tttgtcaggt tcaactccag ccatggtttc
1441 ccagtggagg tcgattctga caccagcatc ttccagctca aggaggtggt tgctaagcga
1501 cagggggttc cggctgacca gttgcgtgtg attttcgcag ggaaggagct gaggaatgac
1561 tggactgtgc agaattgtga cctggatcag cagagcattg ttcacattgt gcagagaccg
1621 tggagaaaag gtcaagaaat gaatgcaact ggaggcgacg accccagaaa cgcggcggga
1681 ggctgtgagc gggagcccca gagcttgact cgggtggacc tcagcagctc agtcctccca
1741 ggagactctg tggggctggc tgtcattctg cacactgaca gcaggaagga ctcaccacca
1801 gctggaagtc cagcaggtag atcaatctac aacagctttt atgtgtattg caaaggcccc
1861 tgtcaaagag tgcagccggg aaaactcagg gtacagtgca gcacctgcag gcaggcaacg
1921 ctcaccttga cccagggtcc atcttgctgg gatgatgttt taattccaaa ccggatgagt
1981 ggtgaatgcc aatccccaca ctgccctggg actagtgcag aatttttctt taaatgtgga
2041 gcacacccca cctctgacaa ggaaacatca gtagctttgc acctgatcgc aacaaatagt
2101 cggaacatca cttgcattac gtgcacagac gtcaggagcc ccgtcctggt tttccagtgc
2161 aactcccgcc acgtgatttg cttagactgt ttccacttat actgtgtgac aagactcaat
2221 gatcggcagt ttgtccacga ccctcagctt ggctactccc tgccttgtgt ggctggctgt
2281 cccaactcct tgattaaaga gctccatcac ttcaggattc tgggagaaga gcagtacaac
2341 cggtaccagc agtatggtgc agaggagtgt gtcctgcaga tggggggcgt gttatgcccc
2401 cgccctggct gtggagcggg gctgctgccg gagcctgacc agaggaaagt cacctgcgaa
2461 gggggcaatg gcctgggctg tgggtttgcc ttctgccggg aatgtaaaga agcgtaccat
2521 gaaggggagt gcagtgccgt atttgaagcc tcaggaacaa ctactcaggc ctacagagtc
2581 gatgaaagag ccgccgagca ggctcgttgg gaagcagcct ccaaagaaac catcaagaaa
2641 accaccaagc cctgtccccg ctgccatgta ccagtggaaa aaaatggagg ctgcatgcac
2701 atgaagtgtc cgcagcccca gtgcaggctc gagtggtgct ggaactgtgg ctgcgagtgg
2761 aaccgcgtct gcatggggga ccactggttc gacgtgtaag gatccaccgg atctagataa
2821 ctgatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca
2881 cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt
2941 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
3001 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta acgcgtaaat
3061 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
3121 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
3181 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
3241 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
3301 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
3361 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
3421 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
3481 cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg
3541 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
3601 ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac
3661 cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
3721 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
3781 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
3841 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
3901 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
3961 aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg
4021 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
4081 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
4141 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
4201 tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt
4261 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
4321 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
4381 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
4441 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
4501 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
4561 ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
4621 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
4681 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
4741 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
4801 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
4861 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
4921 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
4981 atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata
5041 ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg
5101 tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg
5161 agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt
5221 tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca
5281 ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
5341 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
5401 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
5461 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
5521 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
5581 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
5641 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
5701 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
5761 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
5821 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
5881 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
5941 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
6001 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
6061 gccttttgct ggccttttgc tcacatg
//
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