| 质粒名称: | pDisplay-GACh2.0 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 106073 |
| 存储实验室: | Yulong Li |
|
载体骨架基本信息 |
|
| 载体名称: | pDisplay vector |
| 载体出品公司: | Invitrogen |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 7242 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: | -- |
| 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: | GPCR activation based ACh sensor (GACh) 2.0 |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 2049 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | CMV |
| 标签/融合蛋白: | -- |
| 克隆方法: | Gibson克隆 |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | -- |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pDisplay-GACh2.0 was a gift from Yulong Li (Addgene plasmid # 106073 ; http://n2t.net/addgene:106073 ; RRID:Addgene_106073) |
| 参考文献部分: | A genetically encoded fluorescent acetylcholine indicator for in vitro and in vivo studies. Jing M, Zhang P, Wang G, Feng J, Mesik L, Zeng J, Jiang H, Wang S, Looby JC, Guagliardo NA, Langma LW, Lu J, Zuo Y, Talmage DA, Role LW, Barrett PQ, Zhang LI, Luo M, Song Y, Zhu JJ, Li Y. Nat Biotechnol. 2018 Sep;36(8):726-737. doi: 10.1038/nbt.4184. Epub 2018 Jul 9. 10.1038/nbt.4184 PubMed 29985477 |
买家导航
pDisplayGACh2.0载体基本信息
pDisplayGACh2.0载体图谱质粒图谱和多克隆位点信息
pDisplayGACh2.0载体简介
pDisplayGACh2.0载体序列
LOCUS pDisplay-GACh2.0 7242 bp ds-DNA circular SYN 24-OCT-2019
DEFINITION express the genetically-encoded fluorescent ACh indicator GACh2.0 in
mammalian cells.
ACCESSION .
VERSION .
KEYWORDS pDisplay-GACh2.0
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7242)
AUTHORS Jing M, Zhang P, Wang G, Feng J, Mesik L, Zeng J, Jiang H, Wang S,
Looby JC, Guagliardo NA, Langma LW, Lu J, Zuo Y, Talmage DA, Role
LW, Barrett PQ, Zhang LI, Luo M, Song Y, Zhu JJ, Li Y
TITLE A genetically encoded fluorescent acetylcholine indicator for in
vitro and in vivo studies.
JOURNAL Nat Biotechnol. 2018 Sep;36(8):726-737. doi: 10.1038/nbt.4184. Epub
2018 Jul 9.
PUBMED 29985477
REFERENCE 2 (bases 1 to 7242)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Oct 24, 2019 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7242
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 10..389
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 390..593
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 544..564
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 638..657
/label=T7
/note="T7 promoter, forward primer"
promoter 638..656
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 737..799
/codon_start=1
/product="leader sequence from mouse immunoglobulin kappa
light chain"
/label=Ig-kappa leader
/translation="METDTLLLWVLLLWVPGSTGD"
protein_bind 800..824
/gene="mutant version of attB"
/label=attB1
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
primer_bind 1885..1906
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind complement(2012..2033)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(2273..2292)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
CDS 2792..2821
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 2825..2971
/codon_start=1
/product="transmembrane domain from platelet derived growth
factor receptor beta"
/label=PDGFR-beta TM domain
/translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
KKPR"
primer_bind complement(2993..3010)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 2999..3223
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin complement(3355..3943)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3435..3454)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
polyA_signal 4272..4319
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
primer_bind 4344..4363
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
CDS complement(4551..5345)
/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
GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(4662..4681)
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind 5272..5291
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind complement(5444..5463)
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
primer_bind 5690..5710
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
CDS complement(5793..6653)
/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 6416..6435
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6654..6758)
/gene="bla"
/label=AmpR promoter
rep_origin 6785..7240
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(6872..6891)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 7082..7103
/label=F1ori-F
/note="F1 origin, forward primer"
ORIGIN
1 gcgcgcgttg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag
61 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
121 gaccgcccaa cgacccctgc ccattgacgt caataatgac gtatgttccc atagtaacgc
181 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
241 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
301 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
361 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
421 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
481 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
541 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctctctggc
601 taactagaga acccactgct tactggctta tcgaaattaa tacgactcac tatagggaga
661 cccaagcttg gtaccgagct cggatccact agtaacggcc gccagtgtgc tggaattcgg
721 cttggggata tccaccatgg agacagacac actcctgcta tgggtactgc tgctctgggt
781 tccaggttcc actggtgaca caagtttgta caaaaaagtt ggcaccatga ccttgcacaa
841 taacagtaca acctcgcctt tgtttccaaa catcagctcc tcctggatac acagcccctc
901 cgatgcaggg ctgcccccgg gaaccgtcac tcatttcggc agctacaatg tttctcgagc
961 agctggcaat ttctcctctc cagacggtac caccgatgac cctctgggag gtcataccgt
1021 ctggcaagtg gtcttcatcg ctttcttaac gggcatcctg gccttggtga ccatcatcgg
1081 caacatcctg gtaattgtgt catttaaggt caacaagcag ctgaagacgg tcaacaacta
1141 cttcctctta agcctggcct gtgccgatct gattatcggg gtcatttcaa tgaatctgtt
1201 tacgacctac atcatcatga atcgatgggc cttagggaac ttggcctgtg acctctggct
1261 tgccattgac tacgtagcca gcaatgcctc tgttatgaat cttctggtca tcagctttga
1321 cagatacttt tccatcacga ggccgctcac gtaccgagcc aaacgaacaa caaagagagc
1381 cggtgtgatg atcggtctgg cttgggtcat ctcctttgtc ctttgggctc ctgccatctt
1441 gttctggcaa tactttgttg gaaagagaac tgtgcctccg ggagagtgct tcattcagtt
1501 cctcagtgag cccaccatta cttttggcac agccatcgct gctttttata tgcctgtcac
1561 cattatgact attttatact ggagggtctt tcaggaggcc aaaaggcagc tccagaagat
1621 tgacaaatct gagggccgct tccatgtcca gaaccttagc caggtggagc agggaggaaa
1681 cgtctatatc aaggccgaca agcagaagaa cggcatcaag gcgaacttcc acatccgcca
1741 caacatcgag gacggcggcg tgcagctcgc ctaccactac cagcagaaca cccccatcgg
1801 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc gtgcagtcca aactttcgaa
1861 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
1921 cactctcggc atggacgagc tgtacaaggg cggtaccgga gggagcatgg tgagaaaggg
1981 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
2041 ccacaagttc agcgtgtccg gcgagggtga gggcgatgcc acctacggca agctgaccct
2101 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
2161 gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt
2221 caagtccgcc atgcccgaag gctacatcca ggagcgcacc atcttcttca aggacgacgg
2281 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
2341 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
2401 cgctcctagc gtcgcagatg ggcggacggg gcatggactc cgcagatctt ccaagttctg
2461 cttgaaggag cacaaagccc tcaagacgct cagtgcgatc ttgcttgcct tcatcatcac
2521 ttggacccca tacaacatca tggttctggt gaacaccttt tgtgacagct gcatacccaa
2581 aaccttttgg aatctgggct actggctgtg ctacatcaac agcaccgtga accccgtgtg
2641 ctatgctctg tgcaacaaaa cattcagaac cactttcaag atgctgctgc tgtgccagtg
2701 tgacaaaaaa aagaggcgca agcagcagta ccagcagaga cagtcggtca tttttcacaa
2761 gcgcgcaccc gagcaggcct tgtaagtcga cgaacaaaaa ctcatctcag aagaggatct
2821 gaatgctgtg ggccaggaca cgcaggaggt catcgtggtg ccacactcct tgccctttaa
2881 ggtggtggtg atctcagcca tcctggccct ggtggtgctc accatcatct cccttatcat
2941 cctcatcatg ctttggcaga agaagccacg ttaggcggcc gctcgagatc agcctcgact
3001 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
3061 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
3121 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
3181 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
3241 accagtggcg gtaatacggt tatccacaga atcgggggat aacgcaggaa agaacatgtg
3301 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
3361 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
3421 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
3481 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
3541 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
3601 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
3661 tcttgagtcc aacccggtaa gacgcgactt atcgccactg gcagcagcca ctggtaacag
3721 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
3781 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
3841 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
3901 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
3961 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
4021 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
4081 ctaaagtata tatgagtaac ctgaggctat ggcagggcct gccgccccga cgttggctgc
4141 gagccctggg ccttcacccg aacttggggg gtggggtggg gaaaaggaag aaacgcgggc
4201 gtattggccc caatggggtc tcggtggggt atcgacagag tgccagccct gggaccgaac
4261 cccgcgttta tgaacaaacg acccaacacc gtgcgtttta ttctgtcttt ttattgccgt
4321 catagcgcgg gttccttccg gtattgtctc cttccgtgtt tcagttagcc tccccctagg
4381 gtgggcgaag aactccagca tgagatcccc gcgctggagg atcatccagc cggcgtcccg
4441 gaaaacgatt ccgaagccca acctttcata gaaggcggcg gtggaatcga aatctcgtga
4501 tggcaggttg ggcgtcgctt ggtcggtcat ttcgaacccc agagtcccgc tcagaagaac
4561 tcgtcaagaa ggcgatagaa ggcgatgcgc tgcgaatcgg gagcggcgat accgtaaagc
4621 acgaggaagc ggtcagccca ttcgccgcca agctcttcag caatatcacg ggtagccaac
4681 gctatgtcct gatagcggtc cgccacaccc agccggccac agtcgatgaa tccagaaaag
4741 cggccatttt ccaccatgat attcggcaag caggcatcgc catgggtcac gacgagatcc
4801 tcgccgtcgg gcatgctcgc cttgagcctg gcgaacagtt cggctggcgc gagcccctga
4861 tgctcttcgt ccagatcatc ctgatcgaca agaccggctt ccatccgagt acgtgctcgc
4921 tcgatgcgat gtttcgcttg gtggtcgaat gggcaggtag ccggatcaag cgtatgcagc
4981 cgccgcattg catcagccat gatggatact ttctcggcag gagcaaggtg agatgacagg
5041 agatcctgcc ccggcacttc gcccaatagc agccagtccc ttcccgcttc agtgacaacg
5101 tcgagcacag ctgcgcaagg aacgcccgtc gtggccagcc acgatagccg cgctgcctcg
5161 tcttgcagtt cattcagggc accggacagg tcggtcttga caaaaagaac cgggcgcccc
5221 tgcgctgaca gccggaacac ggcggcatca gagcagccga ttgtctgttg tgcccagtca
5281 tagccgaata gcctctccac ccaagcggcc ggagaacctg cgtgcaatcc atcttgttca
5341 atcatgcgaa acgatcctca tcctgtctct tgatcgatct ttgcaaaagc ctaggcctcc
5401 aaaaaagcct cctcactact tctggaatag ctcagaggcc gggcctcggc ctctgcataa
5461 ataaaaaaaa ttagtcagcc atggggcgga gaatgggcgg aactgggcgg agttaggggc
5521 ggaatgggcg gagttagggg cgggactatg gttgctgact aattgagatg catgctttgc
5581 atacttctgc ctgctgggga gcctggggac tttccacacc tggttgctga ctaattgaga
5641 tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca ccctaactga
5701 cacacattcc acagctggtt ctttccgcct caggactctt cctttttcaa taaatcaatc
5761 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
5821 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
5881 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
5941 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
6001 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
6061 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
6121 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
6181 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
6241 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
6301 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
6361 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
6421 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
6481 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
6541 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
6601 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
6661 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
6721 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
6781 cctgacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
6841 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
6901 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
6961 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
7021 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
7081 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
7141 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
7201 tttaacgcga attttaacaa aatattaacg cttacaattt ac
//
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