| 质粒名称: | pcDNA3.1+/mit-2mutAEQ |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 45539 |
| 存储实验室: | Javier Alvarez-Martin |
|
载体骨架基本信息 |
|
| 载体名称: | pcDNA3.1+ |
| 载体出品公司: | Invitrogen |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 5300 bp |
| 完整质粒大小: | 6230 bp |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | G418 |
| 高拷贝/低拷贝: | -- |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | mit-2mutAEQ,Mitochondrially targeted 28,119-double mutated Aequorin |
| 物种来源: | jellyfish |
| 插入大小: | 950 bp |
| 基因突变: | Asp119Ala and Asn28Leu |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | Mitochondrially targeted （插入N端） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | EcoRI(未破坏) |
| 3' 克隆位点: | EcoRI(未破坏) |
| 5' 测序引物 | GAATTCGGCTACGGCTGACCGTTT |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pcDNA3.1+/mit-2mutAEQ was a gift from Javier Alvarez-Martin (Addgene plasmid # 45539 ; http://n2t.net/addgene:45539 ; RRID:Addgene_45539) |
| 参考文献部分: | Mitochondrial free [Ca(2+)] dynamics measured with a novel low-Ca(2+) affinity aequorin probe. de la Fuente S, Fonteriz RI, de la Cruz PJ, Montero M, Alvarez J. Biochem J. 2012 Aug 1;445(3):371-6. doi: 10.1042/BJ20120423. 10.1042/BJ20120423 PubMed 22671130 |
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 pcDNA3.1+/mit-2mutAEQ 6234 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pcDNA3.1+/mit-2mutAEQ
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6234)
AUTHORS de la Fuente S, Fonteriz RI, de la Cruz PJ, Montero M, Alvarez J
TITLE Mitochondrial free [Ca(2+)] dynamics measured with a novel
low-Ca(2+) affinity aequorin probe.
JOURNAL Biochem J. 2012 Aug 1;445(3):371-6. doi: 10.1042/BJ20120423.
PUBMED 22671130
REFERENCE 2 (bases 1 to 6234)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6234
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(44..63)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 235..614
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 615..818
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 769..789
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 863..882
/label=T7
/note="T7 promoter, forward primer"
promoter 863..881
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(1828..1845)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 1834..2058
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 2104..2532
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2191..2210)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2401..2422
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(2541..2561)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 2546..2875
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 2726..2861
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2788..2807
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 2942..3736
/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
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(2996..3015)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3606..3625
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 3910..4031
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3947..3966)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4001..4020
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(4080..4096)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4080..4096)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4093..4115)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4104..4120
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(4128..4158)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4173..4194
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4311..4328)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4482..5067)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4562..4581)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5238..6098)
/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 5861..5880
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6099..6203)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
901 gtttaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattcggc
961 tacggctgac cgttttttgt ggtgtactcc gtgccatcat gtccgtcctg acgccgctgc
1021 tgctgcgggg cttgacaggc tcggcccggc ggctcccagt gccgcgcgcc aagatccatt
1081 cgttgccgcc ggaggggaag ctttatgatg ttcctgatta tgctagcctc aaacttacat
1141 cagacttcga caacccaaga tggattggac gacacaagca tatgttcaat ttccttgatg
1201 tcgaccacct tggaaaaatc tctcttgacg agatggtcta caaggcatct gatattgtca
1261 tcaataacct tggagcaaca cctgagcaag ccaaacgaca caaagatgct gtagaagcct
1321 tcttcggagg agctggaatg aaatatggtg tggaaactga ttggcctgca tatattgaag
1381 gatggaaaaa attggctact gatgaattgg agaaatacgc caaaaacgaa ccaacgctca
1441 tccgtatatg gggtgatgct ttgtttgata tcgtcgacaa agcccaaaat ggagccatta
1501 cactggatga atggaaagca tacaccaaag ctgctggtat catccaatca tcagaagatt
1561 gcgaggaaac attcagagtg tgcgatattg atgaaagtgg acaactcgat gttgatgaga
1621 tgacaagaca acatttagga ttttggtaca ccatggatcc tgcttgcgaa aagctctacg
1681 gtggagctgt cccctaagaa gctctacggt ggtgatgcac cctgggaaga tgatgtgatt
1741 ttgaataaaa cactgatgaa ttctgcagat atccagcaca gtggcggccg ctcgagtcta
1801 gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg
1861 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
1921 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
1981 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg
2041 atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc
2101 cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
2161 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
2221 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
2281 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
2341 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
2401 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
2461 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
2521 ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
2581 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
2641 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
2701 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
2761 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc
2821 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
2881 cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg
2941 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
3001 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
3061 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
3121 gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
3181 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
3241 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
3301 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
3361 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
3421 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga
3481 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
3541 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
3601 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
3661 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
3721 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
3781 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
3841 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
3901 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
3961 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
4021 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg
4081 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
4141 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
4201 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
4261 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
4321 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
4381 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
4441 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
4501 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
4561 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
4621 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
4681 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
4741 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
4801 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
4861 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
4921 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
4981 agctcttgat ccggcaaaca aaccaccgct ggtagcggtt tttttgtttg caagcagcag
5041 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
5101 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
5161 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
5221 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
5281 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
5341 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
5401 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
5461 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
5521 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
5581 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
5641 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
5701 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
5761 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
5821 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
5881 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
5941 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
6001 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
6061 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
6121 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
6181 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtc
//
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