| 质粒名称: | pCMV G-CEPIA1er |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 58215 |
| 存储实验室: |
Masamitsu Iino
|
|
载体骨架基本信息 |
|
| 载体名称: | pCMV/myc/ER |
| 载体出品公司: | Invitrogen |
| 空载体大小: | 4819bp |
| 完整质粒大小: | 6599bp |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 新霉素（G418） |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | Stbl3 |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | G-CEPIA1er、green fluorescent calcium-measuring organelle-entrapped protein indicator for the endoplasmic reticulum |
| 物种来源: | Synthetic |
| 插入大小: | 1480bp |
| 基因突变: | T2A, S13L, A92T, V104T, V114I, E161K, Y193K, F198Y, D202T, I203V, Y258F, A283V, K288T, N290Y relative to RpBphP2 |
| 启动子: | CMV |
| 标签/融合蛋白: | myc (C terminal on insert) |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | None (not destroyed) |
| 3’克隆位点: | XbaI (not destroyed) |
| 5’测序引物 | cgcaaatgggcggtaggcgtg |
| 3’测序引物 | tagaaggcacagtcgagg |
|
文献引用方法 |
|
| 材料和方法部分: | pCMV G-CEPIA1er was a gift from Masamitsu Iino (Addgene plasmid # 58215 ; http://n2t.net/addgene:58215 ; RRID:Addgene_58215) |
| 参考文献部分: | Imaging intraorganellar Ca(2+) at subcellular resolution using CEPIA. Suzuki J, Kanemaru K, Ishii K, Ohkura M, Okubo Y, Iino M. Nat Commun. 2014 Jun 13;5:4153. doi: 10.1038/ncomms5153. 10.1038/ncomms5153 PubMed 24923787 |
买家导航
pCMV G-CEPIA1er载体质粒基本信息
pCMV G-CEPIA1er质粒图谱载体图谱和pCMV G-CEPIA1er载体序列质粒序列多克隆位点信息
pCMV G-CEPIA1er质粒载体简介
pCMV G-CEPIA1er质粒序列载体序列
LOCUS pCMV G-CEPIA1er 6299 bp DNA circular SYN 14-APR-2025
DEFINITION Green fluorescent indicator for calcium imaging in the endoplasmic
reticulum.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6299)
AUTHORS Suzuki J, Kanemaru K, Ishii K, Ohkura M, Okubo Y, Iino M
TITLE Imaging intraorganellar Ca(2+) at subcellular resolution using
CEPIA.
JOURNAL Nat Commun. 2014 Jun 13;5:4153. doi: 10.1038/ncomms5153.
PUBMED 24923787
REFERENCE 2 (bases 1 to 6299)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 14, 2025 from SnapGene Server 8.0.1
https://www.biofeng.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat
Commun."; date: "2014-06-13"; volume: "5:4153. doi"; pages: "
10.1038/ncomms5153"
COMMENT Sequence Label: pCMV G-CEPIA1er
FEATURES Location/Qualifiers
source 1..6299
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 52..428
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 430..633
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 584..604
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
regulatory 690..699
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
sig_peptide join(696..740,823..834)
/codon_start=1
/product="modified signal sequence from the mouse
immunoglobulin heavy chain variable region"
/label=IgH signal sequence
/translation="MGWSCIILFLVATATGAHS"
CDS join(696..741,824..834)
/codon_start=1
/product="signal sequence from the mouse immunoglobulin
heavy chain variableregion"
/label=IgH signal sequence
/translation="MGWSCIILFLVATATGAHS"
CDS 871..927
/codon_start=1
/product="calmodulin-binding peptide from avian smooth
muscle myosin light chain kinase"
/label=calmodulin-binding peptide
/translation="RRKWNKTGHAVRAIGRLSS"
primer_bind 1140..1161
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind complement(1528..1547)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
CDS 1663..2103
/codon_start=1
/product="calcium-binding protein"
/label=calmodulin
/translation="DQLTEEQIAEFKEAFSLFDKDGDGTITTKDLGTVLRSLGQNPTEA
ELQDMINEVDADGNGTIDFPEFLTMMARKMKDTDSEEEIREAFRVWDKDGNGYISAADL
RHVMTNLGEKLTDEEVDEMIREADIDGEGQVNYEEFVQMMTAK"
CDS 2113..2142
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
primer_bind complement(2196..2213)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 2202..2426
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 2472..2900
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2559..2578)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2769..2790
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(2909..2929)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 2914..3243
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 3094..3229
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3156..3175
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3269..4063
/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="MVEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(3323..3342)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3933..3952
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 4237..4358
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4274..4293)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4328..4347
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(4391..4408)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4562..5150)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4642..4661)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(join(5321..6112,6113..6181))
/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 5944..5963
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6182..6286)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 aaagtgccac ctgacgtcag atcgacggat cgggagatcg gtaccgaatt cacattgatt
61 attgagtagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
121 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
181 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
241 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
301 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
361 ccagtacatg accttatggg actttcctac ttggcagtac atctacggtt agtcatcgct
421 attaccatag tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
601 cgtgtacggt gggaggtcta tataagcaga gctttctggc taactagaga acccactgct
661 tactggcacg tggaaattaa tacgacgtgg ccaccatggg atggagctgt atcatcctct
721 tcttggtagc aacagctaca ggtaaggggt taacagtagc aggcttgagg tctggacata
781 tatatgggtg acaatgacat ccactttgcc tttctctcca caggcgcgca ctcccaggtc
841 caactgcagg gatccatggt cgactcttca cgtcgtaagt ggaataagac aggtcacgca
901 gtcagagcta taggtcggct gagctcactc gagaacgtct atatcaaggc cgacgagcag
961 aagaacggca tcaaggcgta cttcaagatc cgccacaaca tcgagggcgg cggcgtgcag
1021 ctcgcctacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
1081 aaccactacc tgagcgtgca gtccatactt tcgaaagacc ccaacgagaa gcgcgatcac
1141 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
1201 aagggcggta ccggagggag catggtgagc aagggcgagg agctgttcac cggggtggtg
1261 cccatccagg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
1321 ggtgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
1381 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc
1441 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
1501 atccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
1561 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
1621 gacggcaaca tcctggggca caagctggag tacaacacgc gtgaccaact gactgaagag
1681 cagatcgcag aatttaaaga ggctttctcc ctatttgaca aggacgggga tgggacgata
1741 acaaccaagg atctggggac ggtgctgcgg tctctggggc agaaccccac agaagcagag
1801 ctccaggaca tgatcaatga agtagatgcc gacggtaatg gcacaatcga cttccctgag
1861 ttcctgacaa tgatggcaag aaaaatgaaa gacacagaca gtgaagaaga aattcgcgaa
1921 gcgttccgtg tgtgggataa ggatggcaat ggctacatct ctgcagcaga ccttcgccac
1981 gtgatgacaa accttggaga gaagttaaca gatgaagagg ttgatgaaat gatcagggaa
2041 gcagatatcg atggagaagg tcaggtaaac tacgaagagt ttgtacaaat gatgacagcg
2101 aaggcggccg cagaacaaaa actcatctca gaagaggatc tgaatggggc cgcaagcgag
2161 aaggacgagc tgtagtctag aagctcgctg atcagcctcg actgtgcctt ctagttgcca
2221 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
2281 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
2341 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
2401 tgctggggat ggcccgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag
2461 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
2521 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
2581 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
2641 gttccgattt agtgctttac ggcacctctc ccccaaaaaa cttgattagg gtgatggttc
2701 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
2761 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
2821 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
2881 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
2941 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
3001 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
3061 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
3121 gcccattctc cgcccctagg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
3181 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
3241 aaaaagctcc ccccccggga ggtccacaat ggttgaacaa gatggattgc acgcaggttc
3301 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
3361 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
3421 cgacctgtcc ggtgccctga atgaactcca ggacgaggca gcgcggctat cgtggctggc
3481 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
3541 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
3601 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
3661 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
3721 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
3781 cgccaggctc aaggcgcgta tgcccgacgg cgaggatctc gtcgtgactc atggcgatgc
3841 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
3901 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
3961 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
4021 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
4081 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
4141 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
4201 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat
4261 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
4321 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgatc
4381 tttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
4441 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
4501 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
4561 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
4621 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
4681 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
4741 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
4801 ccaagctggg ctgtatgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
4861 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
4921 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
4981 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
5041 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
5101 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
5161 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
5221 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
5281 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
5341 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
5401 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
5461 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
5521 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
5581 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
5641 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
5701 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
5761 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
5821 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
5881 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
5941 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
6001 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
6061 gggcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
6121 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
6181 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
6241 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccga
//
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pCMV G-CEPIA1er质粒载体应用举例
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