| 质粒名称: | G-CaMP3 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 22692 |
| 存储实验室: | Loren Looger |
|
载体骨架基本信息 |
|
| 载体名称: | pEGFP-N1 |
| 载体出品公司: | -- |
| 载体抗性: | 卡那霉素 |
| 空载体大小: | 4000 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: | -- |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | G-CaMP3,GCaMP3 |
| 物种来源: | -- |
| 插入大小: | 1300 bp |
| 基因突变: | N363D mutation (increased fluorescence) |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | BglII（销毁后未知） |
| 3' 克隆位点: | NotI（销毁后未知） |
| 5' 测序引物 | EGFP-N |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | G-CaMP3 was a gift from Loren Looger (Addgene plasmid # 22692 ; http://n2t.net/addgene:22692 ; RRID:Addgene_22692) |
| 参考文献部分: | Imaging neural activity in worms, flies and mice with improved GCaMP calcium indicators. Tian L, Hires SA, Mao T, Huber D, Chiappe ME, Chalasani SH, Petreanu L, Akerboom J, McKinney SA, Schreiter ER, Bargmann CI, Jayaraman V, Svoboda K, Looger LL.. Nat Methods. 2009 Dec;6(12):875-81. 10.1038/nmeth.1398 PubMed 19898485 |
买家导航
GCaMP3载体基本信息
GCaMP3载体图谱质粒图谱和多克隆位点信息
GCaMP3载体简介
GCaMP3载体序列
LOCUS G-CaMP3 5311 bp ds-DNA circular SYN 10-APR-2019
DEFINITION a single-wavelength GCaMP2-based genetically encoded calcium
indicator.
ACCESSION .
VERSION .
KEYWORDS G-CaMP3
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5311)
AUTHORS Tian L, Hires SA, Mao T, Huber D, Chiappe ME, Chalasani SH, Petreanu
L, Akerboom J, McKinney SA, Schreiter ER, Bargmann CI, Jayaraman V,
Svoboda K, Looger LL.
TITLE Imaging neural activity in worms, flies and mice with improved GCaMP
calcium indicators.
JOURNAL Nat Methods. 2009 Dec;6(12):875-81.
PUBMED 19898485
REFERENCE 2 (bases 1 to 5311)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 10, 2019 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5311
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 1..204
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 155..175
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
CDS 270..287
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 291..323
/codon_start=1
/product="leader peptide from bacteriophage T7 gene 10"
/label=T7 tag (gene 10 leader)
/note="promotes efficient translation in E. coli"
/translation="MASMTGGQQMG"
CDS 327..350
/codon_start=1
/product="Xpress(TM) epitope tag, including an enterokinase
recognition and cleavage site"
/label=Xpress(TM) tag
/translation="DLYDDDDK"
regulatory 357..366
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 378..434
/codon_start=1
/product="calmodulin-binding peptide from avian smooth
muscle myosin light chain kinase"
/label=calmodulin-binding peptide
/translation="RRKWNKTGHAVRAIGRLSS"
primer_bind 647..668
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind complement(774..795)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(1035..1054)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
CDS 1170..1613
/codon_start=1
/product="calcium-binding protein"
/label=calmodulin
/translation="DQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTEA
ELQDMINEVDADGDGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDGNGYISAAEL
RHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
polyA_signal 1735..1856
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1772..1791)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 1826..1845
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(1863..2318)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2000..2021)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2212..2231
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 2345..2449
/gene="bla"
/label=AmpR promoter
promoter 2451..2808
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(2474..2494)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 2659..2794
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2721..2740
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 2843..3637
/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(2897..2916)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3507..3526
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(3825..3844)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 3869..3916
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 4245..4833
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4734..4753
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
enhancer 5008..5311
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
ORIGIN
1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
181 tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta gcgctaccgg
241 actcagatct cgccaccatg atgggttctc atcatcatca tcatcatggt atggctagca
301 tgactggtgg acagcaaatg ggtcgggatc tgtacgacga tgacgataag gatctcgcca
361 ccatggtcga ctcatcacgt cgtaagtgga ataagacagg tcacgcagtc agagctatag
421 gtcggctgag ctcactcgag aacgtctata tcaaggccga caagcagaag aacggcatca
481 aggcgaactt caagatccgc cacaacatcg aggacggcgg cgtgcagctc gcctaccact
541 accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga
601 gcgtgcagtc caaactttcg aaagacccca acgagaagcg cgatcacatg gtcctgctgg
661 agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag ggcggtaccg
721 gagggagcat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
781 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggt gagggcgatg
841 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
901 ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc
961 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacatc caggagcgca
1021 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
1081 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
1141 tggggcacaa gctggagtac aacacgcgtg accaactgac tgaagagcag atcgcagaat
1201 ttaaagaggc tttctcccta tttgacaagg acggggatgg gacaataaca accaaggagc
1261 tggggacggt gatgcggtct ctggggcaga accccacaga agcagagctg caggacatga
1321 tcaatgaagt agatgccgac ggtgacggca caatcgactt ccctgagttc ctgacaatga
1381 tggcaagaaa aatgaaagac acagacagtg aagaagaaat tagagaagcg ttccgtgtgt
1441 ttgataagga tggcaatggc tacatcagtg cagcagagct tcgccacgtg atgacaaacc
1501 ttggagagaa gttaacagat gaagaggttg atgaaatgat cagggaagca gacatcgatg
1561 gggatggtca ggtaaactac gaagagtttg tacaaatgat gacagcgaag tgagcggccg
1621 cgactctaga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac
1681 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
1741 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
1801 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc
1861 gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
1921 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
1981 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
2041 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
2101 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
2161 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
2221 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
2281 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa
2341 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
2401 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga
2461 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
2521 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
2581 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
2641 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
2701 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
2761 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga
2821 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
2881 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
2941 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
3001 gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac
3061 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
3121 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
3181 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
3241 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
3301 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
3361 gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
3421 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
3481 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
3541 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
3601 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
3661 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
3721 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
3781 gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag
3841 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt
3901 gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc
3961 ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc
4021 ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata
4081 gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
4141 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
4201 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
4261 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
4321 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
4381 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
4441 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
4501 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
4561 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
4621 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
4681 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
4741 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
4801 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
4861 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
4921 gtggataacc gtattaccgc catgcattag ttattaatag taatcaatta cggggtcatt
4981 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
5041 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
5101 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
5161 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
5221 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
5281 catctacgta ttagtcatcg ctattaccat g
//
