| 質粒名稱: | ER-GCaMP6f |
|---|---|
| 出品公司: | Addgene |
| 目錄編號: | 182548 |
| 存儲實驗室: | Vladislav Verkhusha |
|
載體骨架基本信息 |
|
| 載體名稱: | pcDNA 3.1 |
| 載體出品公司: | Clontech |
| 空載體大小: | 5308bp |
| 完整質粒大小: | 5326bp |
| 載體類型: | 哺乳細胞表達, Bacterial Expression |
| 篩選標記: | 新霉素（G418） |
| 載體抗性: | Kanamycin, 50 μg/mL |
| 生長溫度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷貝數: | 未知 |
|
插入基因信息 |
|
| 插入基因名稱: | ER localized P450 |
| 插入大小: | 81bp |
|
文獻引用方法 |
|
| 材料和方法部分: | ER-GCaMP6f was a gift from Chris Richards (Addgene plasmid # 182548 ; //n2t.net/addgene:182548 ; RRID:Addgene_182548) |
| 參考文獻部分: | ER-GCaMP6f: An Endoplasmic Reticulum-Targeted Genetic Probe to Measure Calcium Activity in Astrocytic Processes. Aryal SP, Xia M, Adindu E, Davis C, Ortinski PI, Richards CI. Anal Chem. 2022 Jan 21. doi: 10.1021/acs.analchem.1c04321. 10.1021/acs.analchem.1c04321 PubMed 35061939 |
買家導航
ER-GCaMP6f載體質粒基本信息
ER-GCaMP6f質粒圖譜載體圖譜和ER-GCaMP6f載體序列質粒序列多克隆位點信息
ER-GCaMP6f質粒載體簡介
ER-GCaMP6f質粒序列載體序列
LOCUS ER-GCaMP6f 5326 bp DNA circular SYN 14-APR-2025
DEFINITION Expresses GCaMP6f on the cytosolic side of the ER (P-450site).
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5326)
AUTHORS Aryal SP, Xia M, Adindu E, Davis C, Ortinski PI, Richards CI
TITLE ER-GCaMP6f: An Endoplasmic Reticulum-Targeted Genetic Probe to
Measure Calcium Activity in Astrocytic Processes.
JOURNAL Anal Chem. 2022 Jan 21. doi: 10.1021/acs.analchem.1c04321.
PUBMED 35061939
REFERENCE 2 (bases 1 to 5326)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 14, 2025 from SnapGene Server 8.0.1
//gt12365.cn
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Anal Chem.
2022 Jan 21. doi: 10.1021/acs.analchem.1c04321."
COMMENT Sequence Label: ER-GCaMP6f
FEATURES Location/Qualifiers
source 1..5326
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 112..415
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 416..619
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 570..590
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
regulatory 783..792
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 804..860
/codon_start=1
/product="calmodulin-binding peptide from avian smooth
muscle myosin light chain kinase"
/label=calmodulin-binding peptide
/translation="RRKWNKTGHAVRAIGRLSS"
primer_bind 1073..1094
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind complement(1200..1221)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(1461..1480)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
polyA_signal 2165..2286
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2202..2221)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 2256..2275
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(2293..2748)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2430..2451)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2642..2661
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 2775..2879
/gene="bla"
/label=AmpR promoter
promoter 2881..3238
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
enhancer 2881..3072
/label=SV40 enhancer
/note="enhancer for the SV40 early promoter (Herr, 1993)"
primer_bind complement(2904..2924)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 3089..3224
/label=SV40 ori
/note="SV40 origin of replication"
CDS 3273..4067
/codon_start=1
/product="aminoglycoside phosphotransferase"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
CDS 3273..4067
/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(3327..3346)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3937..3956
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(4255..4274)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 4299..4346
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 4675..5263
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5164..5183
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca ttagttatta
61 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
121 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
181 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
241 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
301 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
361 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
421 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag
481 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
541 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
601 ggtctatata agcagagctg gtttagtgaa ccgtcagatc cgctagcgct accggactca
661 gatctcgagc caccatgcat ccggatatgg accctgtggt cgtgctgggg ctgtgcctga
721 gctgtctgct gctgctgagc ctgtggaagc agtcctacgg aggcgggttg gatccaccgg
781 tcgccaccat ggtcgactca tcacgtcgta agtggaataa gacaggtcac gcagtcagag
841 ctataggtcg gctgagctca ctcgagaacg tctatatcaa ggccgacaag cagaagaacg
901 gcatcaaggc gaacttcaag atccgccaca acatcgagga cggcggcgtg cagctcgcct
961 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
1021 acctgagcgt gcagtccaaa ctttcgaaag accccaacga gaagcgcgat cacatggtcc
1081 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagggcg
1141 gtaccggagg gagcatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
1201 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggtgagg
1261 gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg
1321 tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc
1381 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacatccagg
1441 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
1501 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
1561 acatcctggg gcacaagctg gagtacaacc tgccggacca actgactgaa gagcagatcg
1621 cagaatttaa agaggaattc tccctatttg acaaggacgg ggatgggaca ataacaacca
1681 aggagctggg gacggtgatg cggtctctgg ggcagaaccc cacagaagca gagctgcagg
1741 acatgatcaa tgaagtagat gccgacggtg acggcacaat cgacttccct gagttcctga
1801 caatgatggc aagaaaaatg aaatacaggg acacggaaga agaaattaga gaagcgttcg
1861 gtgtgtttga taaggatggc aatggctaca tcagtgcagc agagcttcgc cacgtgatga
1921 caaaccttgg agagaagtta acagatgaag aggttgatga aatgatcagg gaagcagaca
1981 tcgatgggga tggtcaggta aactacgaag agtttgtaca aatgatgaca gcgaagcttt
2041 aaagcggccg cgactctaga tcataatcag ccataccaca tttgtagagg ttttacttgc
2101 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt
2161 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
2221 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
2281 atcttaaggc gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
2341 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
2401 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
2461 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
2521 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
2581 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
2641 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
2701 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt
2761 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
2821 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc
2881 ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
2941 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
3001 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
3061 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
3121 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
3181 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga
3241 tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
3301 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
3361 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
3421 agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc
3481 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
3541 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
3601 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
3661 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
3721 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
3781 tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg
3841 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
3901 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
3961 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
4021 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
4081 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
4141 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
4201 ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac
4261 acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa
4321 aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc
4381 tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc
4441 accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg
4501 ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta
4561 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
4621 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
4681 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
4741 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
4801 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
4861 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
4921 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
4981 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
5041 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
5101 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
5161 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
5221 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
5281 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttc
//
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