| 质粒名称: | pGP-CMV-Gcamp6m |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 40754 |
| 存储实验室: | Douglas Kim |
|
载体骨架基本信息 |
|
| 载体名称: | pEGFP-N1 |
| 载体出品公司: | Clontech |
| 载体抗性: | 卡那霉素 |
| 空载体大小: | 3955 bp |
| 完整质粒大小: | 5308 bp |
| 载体类型: | 哺乳细胞表达 |
| 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: | GCaMP6m |
| 物种来源: | R. norvegicus (rat), G. gallus (chicken); A. victoria (jellyfish) |
| 插入大小: | 1353 bp |
| 基因又名: |
GCaMP3-T302L R303P M378G K379S D380Y T381R S383T R392G GCaMP3 variant 629 |
| 启动子 | CMV |
| 标签/融合蛋白: |
6xHis (N terminal on insert)
T7 epitope (N terminal on insert)
Xpress tag (N terminal on insert)
|
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | BglII(未破坏) |
| 3' 克隆位点: | NotI(未破坏) |
| 5' 测序引物 | CMV Forward |
| 3' 测序引物 | SV40pA-R |
|
文献引用方法 |
|
| 材料和方法部分: | pGP-CMV-GCaMP6m was a gift from Douglas Kim (爱迪基因 plasmid # 40754) |
| 参考文献部分: | Ultrasensitive fluorescent proteins for imaging neuronal activity. Chen TW, Wardill TJ, Sun Y, Pulver SR, Renninger SL, Baohan A, Schreiter ER, Kerr RA, Orger MB, Jayaraman V, Looger LL, Svoboda K, Kim DS. Nature. 2013 Jul 18;499(7458):295-300. doi: 10.1038/nature12354. 10.1038/nature12354 PubMed 23868258 |
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 pGP-CMV-GCaMP6m 5308 bp ds-DNA circular SYN 10-JAN-2018
DEFINITION Mammalian expression of ultrasensitive protein calcium sensor.
ACCESSION .
VERSION .
KEYWORDS pGP-CMV-GCaMP6m
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5308)
AUTHORS Chen TW, Wardill TJ, Sun Y, Pulver SR, Renninger SL, Baohan A,
Schreiter ER, Kerr RA, Orger MB, Jayaraman V, Looger LL, Svoboda K,
Kim DS
TITLE Ultrasensitive fluorescent proteins for imaging neuronal activity.
JOURNAL Nature. 2013 Jul 18;499(7458):295-300. doi: 10.1038/nature12354.
PUBMED 23868258
COMMENT This file is created by Vector NTI
http://www.biofeng.com/
COMMENT ORIGDB|GenBank
COMMENT VNTAUTHORNAME|biofeng.com|
COMMENT VNTNAME|pGP-CMV-Gcamp6m|
FEATURES Location/Qualifiers
source 1..5308
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 61..364
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 365..568
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 519..539
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
regulatory 616..625
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 622..1974
/codon_start=1
/product="improved fluorescent protein-based calcium sensor
(Chen et al., 2013)"
/label=GCaMP6m
/translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK
TGHAVRAIGRLSSLENVYIKADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDGP
VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE
LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE
LKGIDFKEDGNILGHKLEYNLPDQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRS
LGQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKGSYRDTEEEIREAFGVFDKDGN
GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
regulatory 718..727
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
primer_bind 1008..1029
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind complement(1135..1156)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(1396..1415)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
polyA_signal 2096..2217
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2133..2152)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 2187..2206
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(2224..2679)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2361..2382)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2573..2592
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 2706..2810
/gene="bla"
/label=AmpR promoter
promoter 2812..3169
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(2835..2855)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 3020..3155
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3082..3101
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3204..3998
/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(3258..3277)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3868..3887
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(4186..4205)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 4230..4277
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 4606..5194
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5095..5114
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
601 ccggactcag atctcgccac catgggttct catcatcatc atcatcatgg tatggctagc
661 atgactggtg gacagcaaat gggtcgggat ctgtacgacg atgacgataa ggatctcgcc
721 accatggtcg actcatcacg tcgtaagtgg aataagacag gtcacgcagt cagagctata
781 ggtcggctga gctcactcga gaacgtctat atcaaggccg acaagcagaa gaacggcatc
841 aaggcgaact tcaagatccg ccacaacatc gaggacggcg gcgtgcagct cgcctaccac
901 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
961 agcgtgcagt ccaaactttc gaaagacccc aacgagaagc gcgatcacat ggtcctgctg
1021 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gggcggtacc
1081 ggagggagca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
1141 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg tgagggcgat
1201 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
1261 tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac
1321 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacat ccaggagcgc
1381 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
1441 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
1501 ctggggcaca agctggagta caacctgccg gaccaactga ctgaagagca gatcgcagaa
1561 tttaaagagg ctttctccct atttgacaag gacggggatg ggacaataac aaccaaggag
1621 ctggggacgg tgatgcggtc tctggggcag aaccccacag aagcagagct gcaggacatg
1681 atcaatgaag tagatgccga cggtgacggc acaatcgact tccctgagtt cctgacaatg
1741 atggcaagaa aagggagcta cagggacacg gaagaagaaa ttagagaagc gttcggtgtg
1801 tttgataagg atggcaatgg ctacatcagt gcagcagagc ttcgccacgt gatgacaaac
1861 cttggagaga agttaacaga tgaagaggtt gatgaaatga tcagggaagc agacatcgat
1921 ggggatggtc aggtaaacta cgaagagttt gtacaaatga tgacagcgaa gtgagcggcc
1981 gcgactctag atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa
2041 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
2101 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
2161 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg
2221 cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
2281 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
2341 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
2401 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
2461 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
2521 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
2581 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
2641 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa
2701 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
2761 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg
2821 aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
2881 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
2941 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
3001 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
3061 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
3121 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag
3181 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
3241 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
3301 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
3361 tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga
3421 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
3481 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
3541 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
3601 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
3661 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
3721 ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
3781 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
3841 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
3901 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
3961 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
4021 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
4081 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
4141 ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga
4201 gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg
4261 tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac
4321 cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc
4381 cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat
4441 agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
4501 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
4561 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
4621 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
4681 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
4741 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
4801 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
4861 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
4921 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
4981 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
5041 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
5101 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
5161 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
5221 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
5281 tgtggataac cgtattaccg ccatgcat
//
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