质粒名称: | pCMV(MinDis).iGluSnFR |
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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 |
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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 |
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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)
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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 |
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材料和方法部分: |
pCMV(MinDis).iGluSnFR was a gift from Loren Looger (爱迪基因 plasmid # 41732)
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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 |
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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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