| 质粒名称: | pCMV(MinDis).iGluSnFR |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 41732 |
| 存储实验室: | Loren Looger |
|
载体骨架基本信息 |
|
| 载体名称: | pCMV(MinDis) [variant of pDisplay] |
| 载体出品公司: | modified from Invitrogen |
| 载体抗性: | 氨苄霉素 |
| 空载体大小: | 5300 bp |
| 完整质粒大小: | 6829 bp |
| 载体修饰: | From pDisplay: Hemagglutinin (HA) tag removed |
| 载体类型: | 哺乳细胞表达，谷氨酸生物传感器 |
| 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: | GltI based Glutamate Biosensor |
| 物种来源: | Synthetic; E. coli |
| 插入大小: | 1570 bp |
| 基因又名: | iGluSnFR;gltI/ybeJ;cpGFP146 |
| 基因突变: | cpGFP146 variant from EcMBP165-cpGFP.PPYF.pRSET inserted between AA253-254 of E. coli gltI/ybeJ |
| GenBank编号 | JN247409 |
| 启动子 | CMV |
| 标签/融合蛋白: |
mouse Ig κ-chain (N terminal on backbone)
Myc (C terminal on backbone)
PDGFR transmembrane helix (aa 513-561) (C terminal on backbone)
|
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | BglII(未破坏) |
| 3' 克隆位点: | PstI(未破坏) |
| 5' 测序引物 | CMV-F |
| 3' 测序引物 | BGH-rev |
|
文献引用方法 |
|
| 材料和方法部分: |
pCMV(MinDis).iGluSnFR was a gift from Loren Looger (爱迪基因 plasmid # 41732)
|
| 参考文献部分: | An optimized fluorescent probe for visualizing glutamate neurotransmission. Marvin JS, Borghuis BG, Tian L, Cichon J, Harnett MT, Akerboom J, Gordus A, Renninger SL, Chen TW, Bargmann CI, Orger MB, Schreiter ER, Demb JB, Gan WB, Hires SA, Looger LL. Nat Methods. 2013 Feb;10(2):162-70. doi: 10.1038/nmeth.2333. Epub 2013 Jan 13. 10.1038/nmeth.2333 PubMed 23314171 |
买家导航
pCMV(MinDis).iGluSnFR载体质粒基本信息
pCMV(MinDis).iGluSnFR质粒图谱载体图谱和pCMV(MinDis).iGluSnFR载体序列质粒序列多克隆位点信息
.iGluSnFR质粒图谱.png)
pCMV(MinDis).iGluSnFR质粒载体简介
pCMV(MinDis).iGluSnFR质粒序列载体序列
LOCUS pCMV(MinDis).iGluSnFR 6829 bp ds-DNA circular SYN 10-JAN-2018
DEFINITION a single-wavelength extracellular glutamate sensor constructed from
E. coli Gltl and cpGFP. Membrane-displayed..
ACCESSION .
VERSION .
KEYWORDS pCMV(MinDis).iGluSnFR
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6829)
AUTHORS Marvin JS, Borghuis BG, Tian L, Cichon J, Harnett MT, Akerboom J,
Gordus A, Renninger SL, Chen TW, Bargmann CI, Orger MB, Schreiter
ER, Demb JB, Gan WB, Hires SA, Looger LL
TITLE An optimized fluorescent probe for visualizing glutamate
neurotransmission.
JOURNAL Nat Methods. 2013 Feb;10(2):162-70. doi: 10.1038/nmeth.2333. Epub
2013 Jan 13.
PUBMED 23314171
COMMENT This file is created by Vector NTI
http://www.biofeng.com/
COMMENT ORIGDB|GenBank
COMMENT VNTAUTHORNAME|biofeng.com|
COMMENT VNTNAME|pCMV(MinDis).iGluSnFR|
FEATURES Location/Qualifiers
source 1..6829
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 9..388
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 389..592
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 543..563
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 637..656
/label=T7
/note="T7 promoter, forward primer"
promoter 637..655
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 736..798
/codon_start=1
/product="leader sequence from mouse immunoglobulin kappa
light chain"
/label=Ig-kappa leader
/translation="METDTLLLWVLLLWVPGSTGD"
primer_bind 1764..1785
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind complement(1891..1912)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(2152..2171)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
CDS 2377..2406
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 2410..2556
/codon_start=1
/product="transmembrane domain from platelet derived growth
factor receptor beta"
/label=PDGFR-beta TM domain
/translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
KKPR"
primer_bind complement(2578..2595)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 2584..2808
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin complement(2940..3528)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3020..3039)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
polyA_signal 3857..3904
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
primer_bind 3929..3948
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
CDS complement(4136..4924)
/codon_start=1
/product="aminoglycoside phosphotransferase"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDQEHQGL
APAELFARLKASKPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIALA
TRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(4247..4266)
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind 4851..4870
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
promoter complement(4959..5319)
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 4973..5111
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind complement(5030..5049)
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
primer_bind 5276..5296
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
CDS complement(5379..6239)
/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 6002..6021
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6240..6344)
/gene="bla"
/label=AmpR promoter
rep_origin 6371..6826
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(6458..6477)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 6668..6689
/label=F1ori-F
/note="F1 origin, forward primer"
ORIGIN
1 cgcgcgttga cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt
61 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
121 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
181 aatagggact ttccattgac gtcaatgggt ggactattta cggtaaactg cccacttggc
241 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
301 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
361 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg
421 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
481 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt
541 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagaa ctctctggct
601 aactagagaa cccactgctt actggcttat cgaaattaat acgactcact atagggagac
661 ccaagcttgg taccgaactc ggatccacta gtaacggccg ccagtgtgct ggaattcggc
721 ttggggatat ccaccatgga gacagacaca ctcctgctat gggtactgct gctctgggtt
781 ccaggttcca ctggtgacag atctgccgca ggcagcacgc tggacaaaat cgccaaaaac
841 ggtgtgattg tcgtcggtca ccgtgaatct tcagtgcctt tctcttatta cgacaatcag
901 caaaaagtgg tgggttactc gcaggattac tccaacgcca ttgttgaagc agtgaaaaag
961 aaactcaaca aaccggactt gcaggtaaaa ctgattccga ttacctcaca aaaccgtatt
1021 ccactgctgc aaaacggcac tttcgatttt gaatgtggtt ctaccaccaa caacgtcgaa
1081 cgccaaaaac aggcggcttt ctctgacact attttcgtgg tcggtacgcg cctgttgacc
1141 aaaaagggtg gcgatatcaa agattttgcc aacctgaaag acaaagccgt agtcgtcact
1201 tccggcacta cctctgaagt tttgctcaac aaactgaatg aagagcaaaa aatgaatatg
1261 cgcatcatca gcgccaaaga tcacggtgac tctttccgca ccctggaaag cggtcgtgcc
1321 gttgccttta tgatggatga cgtgctgctg gccggtgaac gtgcgaaagc gaagaaacca
1381 gacaactggg aaatcgtcgg caagccgcag tctcaggagg cctacggttg tatgttgcgt
1441 aaagatgatc cgcagttcaa aaagctgatg gatgacacca tcgctcaggt gcagacctcc
1501 ggtgaagcgg aaaaatggtt tgataagtgg ttcaaaaatc caattctggt gagccacaac
1561 gtctatatca tggccgacaa gcagaggaac ggcatcaagg cgaacttcaa gatccgccac
1621 aacatcgagg acggcggcgt gcagctcgcc taccactacc agcagaacac ccccatcggc
1681 gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccaa actttcgaaa
1741 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
1801 actctcggca tggacgagct gtacaagggc ggtaccggag ggagcatggt gagcaagggc
1861 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
1921 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
1981 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
2041 acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc
2101 aagtccgcca tgcccgaagg ctacatccag gagcgcacca tcttcttcaa ggacgacggc
2161 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
2221 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcataagct ggagtacaac
2281 tttaacaacc ccctgaacat gaatttcgaa ctgtcagacg aaatgaaagc actgttcaaa
2341 gaaccgaatg acaaggcact gaaactgcag gtcgacgaac aaaaactcat ctcagaagag
2401 gatctgaatg ctgtgggcca ggacacgcag gaggtcatcg tggtgccaca ctccttgccc
2461 tttaaggtgg tggtgatctc agccatcctg gccctggtgg tgctcaccat catctccctt
2521 atcatcctca tcatgctttg gcagaagaag ccacgttagg cggccgctcg agatcagcct
2581 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
2641 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
2701 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
2761 attgggaaga caatagcagg cgtgctgggg atgcggtggg ctctatggct tctgaggcgg
2821 aaagaaccag tggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
2881 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
2941 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
3001 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
3061 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
3121 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
3181 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
3241 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
3301 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
3361 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc
3421 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
3481 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
3541 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
3601 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
3661 tcaatctaaa gtatatatga gtaacctgag gctatggcag ggcctgccgc cccgacgttg
3721 gctgcgagcc ctgggccttc acccgaactt ggggggtggg gtggggaaaa ggaagaaacg
3781 cgggcgtatt ggccccaatg gggtctcggt ggggtatcga cagagtgcca gccctgggac
3841 cgaaccccgc gtttatgaac aaacgaccca acaccgtgcg ttttattctg tctttttatt
3901 gccgtcatag cgcgggttcc ttccggtatt gtctccttcc gtgtttcagt tagcctcccc
3961 ctagggtggg cgaagaactc cagcatgaga tccccgcgct ggaggatcat ccagccggcg
4021 tcccggaaaa cgattccgaa gcccaacctt tcatagaagg cggcggtgga atcgaaatct
4081 cgtgatggca ggttgggcgt cgcttggtcg gtcatttcga accccagagt cccgctcaga
4141 agaactcgtc aagaaggcga tagaaggcga tgcgctgcga atcgggagcg gcgataccgt
4201 aaagcacgag gaagcggtca gcccattcgc cgccaagctc ttcagcaata tcacgggtag
4261 ccaacgctat gtcctgatag cggtccgcca cacccagccg gccacagtcg atgaatccag
4321 aaaagcggcc attttccacc atgatattcg gcaagcaggc atcgccatgg gtcacgacga
4381 gatcctcgcc gtcgggcttg ctcgccttga gcctggcgaa cagttcggct ggcgcgagcc
4441 cctgatgctc ttgatcatcc tgatcgacaa gaccggcttc catccgagta cgtgctcgct
4501 cgatgcgatg tttcgcttgg tggtcgaatg ggcaggtagc cggatcaagc gtatgcagcc
4561 gccgcattgc atcagccatg atggatactt tctcggcagg agcaaggtga gatgacagga
4621 gatcctgccc cggcacttcg cccaatagca gccagtccct tcccgcttca gtgacaacgt
4681 cgagcacagc tgcgcaagga acgcccgtcg tggccagcca cgatagccgc gctgcctcgt
4741 cttgcagttc attcagggca ccggacaggt cggtcttgac aaaaagaacc gggcgcccct
4801 gcgctgacag ccggaacacg gcggcatcag agcagccgat tgtctgttgt gcccagtcat
4861 agccgaatag cctctccacc caagcggccg gagaacctgc gtgcaatcca tcttgttcaa
4921 tcatgcgaaa cgatcctcat cctgtctctt gatcgatctt tgcaaaagcc taggcctcca
4981 aaaaagcctc ctcactactt ctggaatagc tcagaggccg aggaggcggc ctcggcctct
5041 gcataaataa aaaaaattag tcagccatgg ggcggagaat gggcggaact gggcggagtt
5101 aggggcggga tgggcggagt taggggcggg actatggttg ctgactaatt gagatgcttg
5161 ctttgcatac ttctgcctgc tggggagcct ggggactttc cacacctggt tgctgactaa
5221 ttgagatgct tgctttgcat acttctgcct gctggggagc ctggggactt tccacaccct
5281 aactgacaca cattccacag ctggttcttt ccgcctcagg actcttcctt tttcaataaa
5341 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
5401 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
5461 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
5521 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
5581 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
5641 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
5701 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
5761 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
5821 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
5881 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
5941 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
6001 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
6061 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
6121 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
6181 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
6241 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
6301 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
6361 gtgccacctg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
6421 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
6481 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
6541 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
6601 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
6661 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
6721 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
6781 caaaaattta acgcgaattt taacaaaata ttaacgctta caatttacg
//
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pCMV(MinDis).iGluSnFR质粒载体应用举例
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