| 质粒名称: | CMV-Mito4x-GCaMP6f |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 127870 |
| 存储实验室: | Timothy Ryan |
|
载体骨架基本信息 |
|
| 载体名称: | pcDNA3 |
| 载体出品公司: | Clontech |
| 空载体大小: | 5433bp |
| 完整质粒大小: | 7197 bp |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 新霉素（G418） |
| 载体抗性: | 氨苄霉素 |
| 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: | 4 repeats of COX8 signal peptide plus GCaMP6f |
| 物种来源: | Synthetic |
| 插入大小: | 1764bp |
| 启动子: | CMV |
| 克隆方法: | 未知 |
| 5’测序引物 | CGCAAATGGGCGGTAGGCGTG |
| 3’测序引物 |
TAGAAGGCACAGTCGAG
(Common Sequencing Primers)
|
|
文献引用方法 |
|
| 材料和方法部分: | CMV-Mito4x-GCaMP6f was a gift from Timothy Ryan (Addgene plasmid # 127870 ; http://n2t.net/addgene:127870 ; RRID:Addgene_127870) |
| 参考文献部分: | Molecular Tuning of the Axonal Mitochondrial Ca(2+) Uniporter Ensures Metabolic Flexibility of Neurotransmission. Ashrafi G, de Juan-Sanz J, Farrell RJ, Ryan TA. Neuron. 2019 Dec 10. pii: S0896-6273(19)30984-5. doi: 10.1016/j.neuron.2019.11.020. 10.1016/j.neuron.2019.11.020 PubMed 31862210 |
买家导航
CMV-Mito4x-GCaMP6f载体质粒基本信息
CMV-Mito4x-GCaMP6f质粒图谱载体图谱和CMV-Mito4x-GCaMP6f载体序列质粒序列多克隆位点信息
CMV-Mito4x-GCaMP6f质粒载体简介
CMV-Mito4x-GCaMP6f质粒序列载体序列
LOCUS CMV-Mito4x-GCaMP6f 7129 bp DNA circular SYN 14-APR-2025
DEFINITION Fluorescent reporter for mitochondrial matrix Ca2+.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7129)
AUTHORS Ashrafi G, de Juan-Sanz J, Farrell RJ, Ryan TA
TITLE Molecular Tuning of the Axonal Mitochondrial Ca(2+) Uniporter
Ensures Metabolic Flexibility of Neurotransmission.
JOURNAL Neuron. 2019 Dec 10. pii: S0896-6273(19)30984-5. doi:
10.1016/j.neuron.2019.11.020.
PUBMED 31862210
REFERENCE 2 (bases 1 to 7129)
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: "Neuron.
2019 Dec 10. pii: S0896-6273(19)30984-5. doi:
10.1016/j.neuron.2019.11.020."
COMMENT Sequence Label: CMV-Mito4x-GCaMP6f
FEATURES Location/Qualifiers
source 1..7129
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS 75..869
/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(129..148)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 739..758
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 1043..1164
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1080..1099)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 1134..1153
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(1213..1229)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(1213..1229)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(1226..1248)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 1237..1253
/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(join(1261..1267,1268..1285,1286..1291))
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 1306..1327
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(1444..1461)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(1615..2203)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(1695..1714)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(join(2374..3165,3166..3234))
/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 2997..3016
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(3235..3339)
/gene="bla"
/label=AmpR promoter
primer_bind complement(3414..3433)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 3605..3984
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 3985..4188
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 4139..4159
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 4233..4252
/label=T7
/note="T7 promoter, forward primer"
promoter 4233..4251
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 4283..4357
/codon_start=1
/product="mitochondrial presequence of human cytochrome c
oxidase subunit VIII (Rizutto et al., 1989)"
/label=COX8 presequence
/translation="MSVLTPLLLRGLTGSARRLPVPRAK"
CDS 4379..4453
/codon_start=1
/product="mitochondrial presequence of human cytochrome c
oxidase subunit VIII (Rizutto et al., 1989)"
/label=COX8 presequence
/translation="MSVLTPLLLRGLTGSARRLPVPRAK"
CDS 4475..4549
/codon_start=1
/product="mitochondrial presequence of human cytochrome c
oxidase subunit VIII (Rizutto et al., 1989)"
/label=COX8 presequence
/translation="MSVLTPLLLRGLTGSARRLPVPRAK"
CDS 4571..4645
/codon_start=1
/product="mitochondrial presequence of human cytochrome c
oxidase subunit VIII (Rizutto et al., 1989)"
/label=COX8 presequence
/translation="MSVLTPLLLRGLTGSARRLPVPRAK"
CDS 4703..4720
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS join(4760..4768,4769..4783)
/codon_start=1
/product="Xpress(TM) epitope tag, including an enterokinase
recognition and cleavage site"
/label=Xpress(TM) tag
/translation="DLYDDDDK"
CDS 4811..4867
/codon_start=1
/product="calmodulin-binding peptide from avian smooth
muscle myosin light chain kinase"
/label=calmodulin-binding peptide
/translation="RRKWNKTGHAVRAIGRLSS"
primer_bind complement(5468..5487)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind complement(6090..6107)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 6096..6320
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 6366..6794
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(6453..6472)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 6663..6684
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(6803..6823)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter join(6808..7129,1..8)
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 6988..7123
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 7050..7069
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
ORIGIN
1 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat
61 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
121 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
181 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
241 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
301 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
361 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
421 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
481 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
541 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
601 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
661 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
721 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
781 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
841 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca
901 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
961 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
1021 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag
1081 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
1141 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt
1201 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
1261 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
1321 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
1381 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
1441 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
1501 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
1561 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
1621 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
1681 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
1741 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
1801 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
1861 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
1921 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
1981 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
2041 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
2101 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
2161 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
2221 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
2281 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
2341 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
2401 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
2461 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
2521 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
2581 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
2641 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
2701 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
2761 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
2821 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
2881 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
2941 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
3001 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
3061 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
3121 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
3181 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
3241 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
3301 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
3361 acctgacgtc gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc
3421 tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct
3481 gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg
3541 aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg
3601 cgttgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
3661 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
3721 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
3781 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
3841 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
3901 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
3961 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
4021 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
4081 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
4141 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact
4201 agagaaccca ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa
4261 gctggctagc ggatccaccg ccatgagcgt gctgacacct ctgctgctga gaggcctgac
4321 aggctctgct agaaggctgc ctgtgcctag agccaagatc cactctctgg gcgaccctat
4381 gtctgtgctg actccactgc tcctgcgggg actgactgga tctgctcgta gactgccagt
4441 gccacgggct aagattcaca gcctgggcga tccaatgagt gtgcttaccc cactcctgct
4501 cagaggactg accggaagtg ctagacgcct gccagttcca agggccaaaa tccatagtct
4561 gggagatccc atgtccgtgt tgacacccct cttgctgagg ggacttaccg gctcagctcg
4621 gagactccct gttccaagag caaagattca ttccctcggc gacccaagat ctggcagcgc
4681 taaagaccct accatgggta gccaccacca ccatcaccac ggcatggctt ctatgacagg
4741 cggccagcag atgggccgcg acctgtacga tgacgacgat aaggacctgg ccacaatggt
4801 ggacagcagc aggcggaagt ggaacaagac aggccacgcc gttagagcca tcggcagact
4861 gtctagcctg gaaaacgtgt acatcaaggc cgacaagcag aagaacggca tcaaggctaa
4921 cttcaagatc cggcacaaca tcgaggacgg cggagtgcag ctggcttacc actaccagca
4981 gaacacccct atcggcgacg gaccagttct gctgcctgac aaccactacc tgagcgtgca
5041 gagcaagctg agcaaggacc ccaacgagaa gagggaccac atggtgctgc tggaatttgt
5101 gaccgccgct ggcatcacac tcggcatgga cgaactgtac aaaggcggca caggcggctc
5161 tatggtgtcc aagggcgaag aactgttcac cggcgtggtg cccattctgg tggaactgga
5221 tggggatgtg aacggccaca agttcagcgt cagcggagaa ggcgaaggcg acgccacata
5281 cggaaagctg accctgaagt tcatctgcac caccggcaaa ctgcccgtgc cttggcctac
5341 actggtcacc acactgacat acggcgtgca gtgcttcagc agataccccg accatatgaa
5401 gcagcacgac ttcttcaaga gcgccatgcc tgagggctac atccaagaga gaaccatctt
5461 ctttaaggac gacggcaact acaagaccag ggccgaagtg aagttcgagg gcgacaccct
5521 ggtcaacaga atcgagctga agggcatcga cttcaaagag gacggcaaca tcctgggcca
5581 caagctcgag tacaacctgc ctgaccagct gaccgaggaa cagatcgccg agttcaaaga
5641 agagttcagc ctgttcgaca aggacggcga cggcacaatc accaccaaag aactgggcac
5701 cgtgatgaga agcctgggac agaaccctac agaggccgag ctgcaggaca tgatcaacga
5761 ggttgacgct gatggcgacg gaactatcga cttccccgag ttcctgacca tgatggccag
5821 aaagatgaag tacagggaca cagaggaaga gatccgcgag gccttcggcg tgttcgataa
5881 ggatggaaac ggctacatct ctgccgctga gctgagacac gtgatgacca acctgggcga
5941 gaagctgaca gacgaagagg tggacgagat gatcagagag gccgacattg acggcgacgg
6001 gcaagtgaac tacgaagagt ttgtccagat gatgaccgcc aagtgaaccg gtgtcgacga
6061 attcgggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc
6121 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
6181 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
6241 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
6301 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta
6361 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
6421 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
6481 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
6541 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
6601 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
6661 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
6721 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
6781 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
6841 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
6901 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
6961 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
7021 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc
7081 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggct
//
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CMV-Mito4x-GCaMP6f质粒载体应用举例
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