| 质粒名称: | pcDNA3 AIF 1-90 mCherry FRB |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 67530 |
| 存储实验室: | Stephen Tait |
|
载体骨架基本信息 |
|
| 载体名称: | pcDNA3 |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 5500 bp |
| 完整质粒大小: | 6687 bp |
| 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: | AIF a.a. 1 - 90 |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 270 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | CMV |
| 标签/融合蛋白: | mCherry（插入件上的C端子） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | Hind III（销毁后未知） |
| 3' 克隆位点: | Kpn I（销毁后未知） |
| 5' 测序引物 | CMV |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pcDNA3 AIF 1-90 mCherry FRB was a gift from Stephen Tait (Addgene plasmid # 67530 ; http://n2t.net/addgene:67530 ; RRID:Addgene_67530) |
| 参考文献部分: | Limited mitochondrial permeabilization causes DNA damage and genomic instability in the absence of cell death. Ichim G, Lopez J, Ahmed SU, Muthalagu N, Giampazolias E, Delgado ME, Haller M, Riley JS, Mason SM, Athineos D, Parsons MJ, van de Kooij B, Bouchier-Hayes L, Chalmers AJ, Rooswinkel RW, Oberst A, Blyth K, Rehm M, Murphy DJ, Tait SW. Mol Cell. 2015 Mar 5;57(5):860-72. doi: 10.1016/j.molcel.2015.01.018. Epub 2015 Feb 19. 10.1016/j.molcel.2015.01.018 PubMed 25702873 |
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 pcDNA3 AIF 1-90 mCherry FRB 6687 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION For expression of mCherry fused FRB to the mitochondrial
innermembrane.
ACCESSION .
VERSION .
KEYWORDS pcDNA3 AIF 1-90 mCherry FRB
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6687)
AUTHORS Ichim G, Lopez J, Ahmed SU, Muthalagu N, Giampazolias E, Delgado ME,
Haller M, Riley JS, Mason SM, Athineos D, Parsons MJ, van de Kooij
B, Bouchier-Hayes L, Chalmers AJ, Rooswinkel RW, Oberst A, Blyth K,
Rehm M, Murphy DJ, Tait SW
TITLE Limited mitochondrial permeabilization causes DNA damage and genomic
instability in the absence of cell death.
JOURNAL Mol Cell. 2015 Mar 5;57(5):860-72. doi:
10.1016/j.molcel.2015.01.018. Epub 2015 Feb 19.
PUBMED 25702873
REFERENCE 2 (bases 1 to 6687)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6687
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(44..63)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 235..614
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 615..818
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 769..789
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 863..882
/label=T7
/note="T7 promoter, forward primer"
promoter 863..881
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 1168..1197
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 1204..1908
/codon_start=1
/product="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
/label=mCherry
/note="mammalian codon-optimized"
/translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
GRHSTGGMDELYK"
primer_bind complement(1346..1364)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind 1614..1633
/label=mCherry-F
/note="mCherry, forward primer"
CDS 1930..2208
/codon_start=1
/product="T2098L mutant of FKBP-rapamycin binding domain of
human FRAP"
/label=FRB*
/note="binds synthetic ligands such as AP21967"
/translation="ILWHEMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTL
KETSFNQAYGRDLMEAQEWCRKYMKSGNVKDLLQAWDLYYHVFRRISK"
promoter complement(2239..2257)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(2240..2257)
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind complement(2277..2294)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 2283..2507
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 2553..2981
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2640..2659)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2850..2871
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(2990..3010)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 2995..3325
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 3176..3311
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3238..3257
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3392..4186
/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
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(3446..3465)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 4056..4075
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 4360..4481
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4397..4416)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4451..4470
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(4530..4546)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4530..4546)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4543..4565)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4554..4570
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(4578..4608)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4623..4644
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4761..4778)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4932..5520)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5012..5031)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5691..6551)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 6314..6333
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6552..6656)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttatgttc
901 cggtgtggag gcctggcggc gggtgctttg aagcagaagc tggtgccctt ggtgcggacc
961 gtgtgcgtcc gaagcccgag gcagaggaac cggctcccag gcaacttgtt ccagcgatgg
1021 catgttcctc tagaactcca gatgacaaga caaatggcta gctctggtgc atcagggggc
1081 aaaatcgata attctgtgtt agtccttatt gtgggcttat caacagtagg agctggtgcc
1141 tatgcctaca agactatgaa agagatggaa caaaaactta tttctgaaga agatctgggt
1201 accgtgagca agggcgagga ggataacatg gccatcatca aggagttcat gcgcttcaag
1261 gtgcacatgg agggctccgt gaacggccac gagttcgaga tcgagggcga gggcgagggc
1321 cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc
1381 ttcgcctggg acatcctgtc ccctcagttc atgtacggct ccaaggccta cgtgaagcac
1441 cccgccgaca tccccgacta cttgaagctg tccttccccg agggcttcaa gtgggagcgc
1501 gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc aggactcctc cctgcaggac
1561 ggcgagttca tctacaaggt gaagctgcgc ggcaccaact tcccctccga cggccccgta
1621 atgcagaaga agaccatggg ctgggaggcc tcctccgagc ggatgtaccc cgaggacggc
1681 gccctgaagg gcgagatcaa gcagaggctg aagctgaagg acggcggcca ctacgacgct
1741 gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc ctacaacgtc
1801 aacatcaagt tggacatcac ctcccacaac gaggactaca ccatcgtgga acagtacgaa
1861 cgcgccgagg gccgccactc caccggcggc atggacgagc tgtacaaggc ggccgccgga
1921 ggtggcggga tcctctggca tgagatgtgg catgaaggcc tggaagaggc atctcgtttg
1981 tactttgggg aaaggaacgt gaaaggcatg tttgaggtgc tggagccctt gcatgctatg
2041 atggaacggg gcccccagac tctgaaggaa acatccttta atcaggccta tggtcgagat
2101 ttaatggagg cccaagagtg gtgcaggaag tacatgaaat cagggaatgt caaggacctc
2161 ctccaagcct gggacctcta ttatcatgtg ttccgacgaa tctcaaagtg actcgagcat
2221 gcatctagag ggccctattc tatagtgtca cctaaatgct agagctcgct gatcagcctc
2281 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
2341 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
2401 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
2461 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga
2521 aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
2581 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
2641 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
2701 aaatcggggc atccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
2761 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
2821 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
2881 caaccctatc tcggtctatt cttttgattt ataagggatt ttggggattt cggcctattg
2941 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt
3001 cagttagggt gtggaaagtc cccaggctcc ccaggcaggc agaagtatgc aaagcatgca
3061 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
3121 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
3181 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
3241 ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt
3301 ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc
3361 tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
3421 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
3481 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
3541 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct
3601 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
3661 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
3721 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
3781 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
3841 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
3901 gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
3961 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
4021 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
4081 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
4141 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
4201 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
4261 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
4321 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat
4381 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
4441 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg
4501 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
4561 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
4621 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
4681 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
4741 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
4801 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
4861 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
4921 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
4981 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
5041 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
5101 ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag
5161 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
5221 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
5281 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
5341 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
5401 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
5461 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
5521 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
5581 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
5641 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
5701 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
5761 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
5821 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
5881 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
5941 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
6001 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
6061 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
6121 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
6181 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
6241 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
6301 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
6361 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
6421 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
6481 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
6541 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
6601 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
6661 tttccccgaa aagtgccacc tgacgtc
//
