| 质粒名称: | CMV-mito-GEM-GECO1 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 32461 |
| 存储实验室: | Robert Campbell |
|
载体骨架基本信息 |
|
| 载体名称: | pcDNA3.1 (-) |
| 载体出品公司: | Invitrogen |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 5600 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: | -- |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | GEM-GECO,emission ratiometric genetically encoded Ca2+-indicators for optical imaging |
| 物种来源: | -- |
| 插入大小: | 1248 bp |
| 基因突变: | GCaMP3 L60P/K69E/N77Y/D86G/N98I/K119I/L173Q/T223S/N302S/R377P/K380Q/S 404G/E430V |
| GenBank编号 | JN258409 |
| 启动子 | CMV |
| 标签/融合蛋白: | 细胞色素c氧化酶亚基VIII（主干N端）线粒体靶向信号的双重性 |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | BamHI(未破坏) |
| 3' 克隆位点: | HindIII(未破坏) |
| 5' 测序引物 | TAATACGACTCACTATAGGG |
| 3' 测序引物 | TAGAAGGCACAGTCGAGG |
|
文献引用方法 |
|
| 材料和方法部分: | CMV-mito-GEM-GECO1 was a gift from Robert Campbell (Addgene plasmid # 32461 ; http://n2t.net/addgene:32461 ; RRID:Addgene_32461) |
| 参考文献部分: | An Expanded Palette of Genetically Encoded Ca2+ Indicators. Zhao Y, Araki S, Wu J, Teramoto T, Chang YF, Nakano M, Abdelfattah AS, Fujiwara M, Ishihara T, Nagai T, Campbell RE. Science. 2011 Sep 8. 10.1126/science.1208592 PubMed 21903779 |
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 CMV-mito-GEM-GECO1 6815 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Mammalian expression of mitochondria-targeting blue-green emission
ratiometric genetically encoded Ca2+ indicator for optical imaging.
ACCESSION .
VERSION .
KEYWORDS CMV-mito-GEM-GECO1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6815)
AUTHORS Zhao Y, Araki S, Wu J, Teramoto T, Chang YF, Nakano M, Abdelfattah
AS, Fujiwara M, Ishihara T, Nagai T, Campbell RE
TITLE An Expanded Palette of Genetically Encoded Ca2+ Indicators.
JOURNAL Science. 2011 Sep 8.
PUBMED 21903779
REFERENCE 2 (bases 1 to 6815)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6815
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(49..68)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 240..619
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 620..823
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 774..794
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 868..887
/label=T7
/note="T7 promoter, forward primer"
promoter 868..886
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 1152..1208
/codon_start=1
/product="calmodulin-binding peptide from avian smooth
muscle myosin light chain kinase"
/label=calmodulin-binding peptide
/translation="RRKWNKTGHAVRAIGRLSS"
primer_bind 1421..1442
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind complement(1806..1825)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind complement(2411..2428)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 2417..2641
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 2687..3115
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2774..2793)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2984..3005
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(3124..3144)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 3129..3458
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 3309..3444
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3371..3390
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3525..4319
/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(3579..3598)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 4189..4208
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 4493..4614
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4530..4549)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4584..4603
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(4663..4679)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4663..4679)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4676..4698)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4687..4703
/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(4711..4741)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4756..4777
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4894..4911)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5065..5653)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5145..5164)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5824..6684)
/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 6447..6466
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6685..6789)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 acgtcgacgg atcgggagat ctcccgatcc cctatggtgc actctcagta caatctgctc
61 tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta
121 gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa
181 tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg
241 acattgatta ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc
301 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
361 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
421 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
481 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
541 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
601 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
661 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
721 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
781 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga
841 acccactgct tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg
901 ctagcgttta aacgggccct ctagactcga gcatgtctgt tctgactcct ctgctgctcc
961 ggggtctcac aggttccgca agaagactcc ccgtgcctag ggccaaaatt cattcactgg
1021 gggaccccat gagcgtgctc accccactcc tgctgcgggg gctgaccggc agcgctaggc
1081 ggctgccagt ccccagggcc aagatccaca gtctcggcga tcccaaggat ccaaccatgg
1141 tcgactcatc acgtcgtaag tggaataaga caggtcacgc agtcagagct ataggtcggc
1201 tgagctcacc agagaacgtg tatataaagg ccgacgagca gaagaacggc atcaaggcgt
1261 acttcaagat ccgccacaac atcgagggcg gcggcgtgca gctcgcctac cactaccagc
1321 agatcacccc cataggcgac ggccccgtgc tgctgcccga caaccactac ctgagcgtgc
1381 agtccatact ttcgaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
1441 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagggtggc agcggtggca
1501 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catccaggtc gagctggacg
1561 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg tgagggcgat gccacctacg
1621 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
1681 tcgtgaccac cctgtcctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc
1741 agcacgactt cttcaagtcc gccatgcccg aaggctacat ccaggagcgc accatcttct
1801 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
1861 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
1921 agctggagta cagcacgcgt gaccaactga ctgaagagca gatcgcagaa tttaaagagg
1981 ctttctccct atttgacaag gacggggatg ggacgataac aaccaaggag ctggggacgg
2041 tgatgcggtc tctggggcag aaccccacag aagcagagct gcaggacatg atcaatgaag
2101 tagatgccga cggtgacggc acaatcgact tccctgagtt cctgacaatg atggcaccta
2161 aaatgcagga cacagacagt gaagaagaaa ttagagaagc gttccgtgtg tttgataagg
2221 acggcaatgg ctacatcggc gcagcagagc ttcgccacgt gatgacaaac cttggagaga
2281 agttaacaga tgaagaggtt gatgaaatga tcagggttgc agacatcgat ggggatggtc
2341 aggtaaacta cgaagagttt gtacaaatga tgacagcgaa gtagaagctt aagtttaaac
2401 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
2461 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
2521 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
2581 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
2641 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc
2701 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
2761 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
2821 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
2881 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
2941 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
3001 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
3061 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
3121 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
3181 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
3241 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
3301 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
3361 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
3421 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
3481 ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
3541 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
3601 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
3661 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc
3721 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
3781 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
3841 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
3901 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
3961 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
4021 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg
4081 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
4141 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
4201 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
4261 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
4321 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
4381 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
4441 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
4501 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
4561 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
4621 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
4681 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
4741 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
4801 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
4861 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
4921 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
4981 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
5041 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
5101 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
5161 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
5221 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
5281 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
5341 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
5401 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
5461 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
5521 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
5581 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
5641 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
5701 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
5761 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
5821 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
5881 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
5941 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
6001 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
6061 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
6121 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
6181 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
6241 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
6301 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
6361 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
6421 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
6481 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
6541 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
6601 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
6661 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
6721 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
6781 ggttccgcgc acatttcccc gaaaagtgcc acctg
//
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