| 质粒名称: | pCS2+HyPer7 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 136466 |
| 存储实验室: | Vsevolod Belousov |
|
载体骨架基本信息 |
|
| 载体名称: | pCS2+ |
| 完整质粒大小: | 5531 bp |
| 载体类型: | 哺乳细胞表达 |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | HyPer7 |
| 插入大小: | 1440 bp |
| 启动子: | CMV, SP6 |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | EcoRI(未破坏) |
| 3’克隆位点: | XhoI(未破坏) |
| 5’测序引物 | ATTTAGGTGACACTATAGA |
| 3’测序引物 | GTCATAGCTGTTTCCTG |
|
文献引用方法 |
|
| 材料和方法部分: | pCS2+HyPer7 was a gift from Vsevolod Belousov (Addgene plasmid # 136466 ; http://n2t.net/addgene:136466 ; RRID:Addgene_136466) |
| 参考文献部分: | Ultrasensitive Genetically Encoded Indicator for Hydrogen Peroxide Identifies Roles for the Oxidant in Cell Migration and Mitochondrial Function. Pak VV, Ezerina D, Lyublinskaya OG, Pedre B, Tyurin-Kuzmin PA, Mishina NM, Thauvin M, Young D, Wahni K, Martinez Gache SA, Demidovich AD, Ermakova YG, Maslova YD, Shokhina AG, Eroglu E, Bilan DS, Bogeski I, Michel T, Vriz S, Messens J, Belousov VV. Cell Metab. 2020 Mar 3;31(3):642-653.e6. doi: 10.1016/j.cmet.2020.02.003. 10.1016/j.cmet.2020.02.003 PubMed 32130885 |
买家导航
pCS2HyPer7载体基本信息
pCS2HyPer7载体图谱质粒图谱和多克隆位点信息
pCS2HyPer7载体简介
pCS2HyPer7载体序列
LOCUS pCS2+HyPer7 5531 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Mammalian expression of non-targeted ultrasensitive hydrogen
peroxide indicator HyPer7 for optical imaging.
ACCESSION .
VERSION .
KEYWORDS pCS2+HyPer7
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5531)
AUTHORS Pak VV, Ezerina D, Lyublinskaya OG, Pedre B, Tyurin-Kuzmin PA,
Mishina NM, Thauvin M, Young D, Wahni K, Martinez Gache SA,
Demidovich AD, Ermakova YG, Maslova YD, Shokhina AG, Eroglu E, Bilan
DS, Bogeski I, Michel T, Vriz S, Messens J, Belousov VV
TITLE Ultrasensitive Genetically Encoded Indicator for Hydrogen Peroxide
Identifies Roles for the Oxidant in Cell Migration and Mitochondrial
Function.
JOURNAL Cell Metab. 2020 Mar 3;31(3):642-653.e6. doi:
10.1016/j.cmet.2020.02.003.
PUBMED 32130885
REFERENCE 2 (bases 1 to 5531)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5531
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 35..53
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind 35..52
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind 717..738
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind complement(847..868)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(1108..1127)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
promoter complement(1561..1578)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
polyA_signal 1583..1717
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1607..1626)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 1661..1680
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind complement(1830..1850)
/label=T3
/note="T3 promoter, forward primer"
promoter complement(1830..1848)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(1869..1885)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(1869..1885)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(1882..1904)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 1893..1909
/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(1917..1947)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 1962..1983
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(2100..2117)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(2271..2859)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(2351..2370)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(3030..3890)
/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 3653..3672
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(3891..3995)
/gene="bla"
/label=AmpR promoter
rep_origin complement(4021..4476)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4158..4179)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 4370..4389
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 4554..5531
/label=CMV IE94 promoter
/note="enhancer/promoter region of simian cytomegalovirus
major immediate early transcription unit IE94"
ORIGIN
1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
61 ctacttgttc tttttgcagg atcccatcga ttcgaattcg ccaccatgca cctggctaat
121 gaggagcaaa acgaattgga aggtgctttt aagctgggct tgatctttac cgtcgccccg
181 tacttgctgc ctaagttgat cgtcagcctg cgccgcaccg ccccgaagat gcctttgatg
241 ttggaggaga attacaccca cacccttacc gagtcactga aacgcggcga cgttgacgcc
301 atcattgtgg cagagccgtt tcaagagccg ggtattgtga ctgaaccttt gtatgacgag
361 ccgtttttcg ttatcgttcc gaagggccat agcttcgagg agttggatgc cgttagccct
421 cgcatgttgg gcgaagaaca agttctgctt ctgaccgagg gcaactgtat gcgtgaccag
481 gtgcttagct ccgccggctt caacagcaac aacgtctata tcatggccga caagcagaag
541 aacggcatca aggccaactt caagatccgc cacaacgtcg aggacggcag cgtgcagctc
601 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
661 cactacctga gcttccagtc cgtcctgagc aaagacccca acgagaagcg cgatcacatg
721 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaac
781 gtggatggcg gtagcggtgg caccggcagc aagggcgagg agctgttcac cggggtggtg
841 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
901 ggcgagggcg atgccaccta cggcaagctg accctgaagc tgatctgcac caccggcaag
961 ctgcccgtgc cctggcccac cctcgtgacc accctcagct acggcctgaa gtgcttcgcc
1021 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
1081 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
1141 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcgg cttcaaggag
1201 gacggcaaca tcctggggca caagctggag tacaacggcc ccagttgcag cgagcttgcc
1261 gccaaacagc gcattcaggg tctcaccaac acccttcaag gctcaagcat caataccatc
1321 cgccacatgg ttgccagtgg cttggcaatc agcgtcttgc ctgccactgc ccttaccgaa
1381 aacgaccaca tgctgtttag cattatcccg tttgaaggca ccccgcctag ccgccgcgtt
1441 gtcctggcct atcgccgcaa ctttgtccgc cctaaggccc tttcagccat gaaagccgca
1501 attatgcaaa gccaactgca tggtgttagc ttcatctgcg attgactcga gcctctagaa
1561 ctatagtgag tcgtattacg tagatccaga catgataaga tacattgatg agtttggaca
1621 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
1681 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
1741 tatgtttcag gttcaggggg aggtgtggga ggttttttaa ttcgcggccg cggcgccaat
1801 gcattgggcc cggtacccag cttttgttcc ctttagtgag ggttaattgc gcgcttggcg
1861 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
1921 atacgagccg ggagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
1981 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
2041 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
2101 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
2161 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
2221 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
2281 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
2341 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
2401 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
2461 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
2521 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
2581 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
2641 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
2701 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
2761 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
2821 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
2881 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
2941 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
3001 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
3061 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
3121 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
3181 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
3241 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
3301 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
3361 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
3421 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
3481 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
3541 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
3601 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
3661 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
3721 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
3781 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
3841 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
3901 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
3961 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
4021 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
4081 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
4141 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
4201 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
4261 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
4321 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
4381 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
4441 cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc aggctgcgca
4501 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagtcg accgccaatt
4561 caatatggcg tatatggact catgccaatt caatatggtg gatctggacc tgtgccaatt
4621 caatatggcg tatatggact cgtgccaatt caatatggtg gatctggacc ccagccaatt
4681 caatatggcg gacttggcac catgccaatt caatatggcg gacctggcac tgtgccaact
4741 ggggaggggt ctacttggca cggtgccaag tttgaggagg ggtcttggcc ctgtgccaag
4801 tccgccatat tgaattggca tggtgccaat aatggcggcc atattggcta tatgccagga
4861 tcaatatata ggcaatatcc aatatggccc tatgccaata tggctattgg ccaggttcaa
4921 tactatgtat tggccctatg ccatatagta ttccatatat gggttttcct attgacgtag
4981 atagcccctc ccaatgggcg gtcccatata ccatatatgg ggcttcctaa taccgcccat
5041 agccactccc ccattgacgt caatggtctc tatatatggt ctttcctatt gacgtcatat
5101 gggcggtcct attgacgtat atggcgcctc ccccattgac gtcaattacg gtaaatggcc
5161 cgcctggctc aatgcccatt gacgtcaata ggaccaccca ccattgacgt caatgggatg
5221 gctcattgcc cattcatatc cgttctcacg ccccctattg acgtcaatga cggtaaatgg
5281 cccacttggc agtacatcaa tatctattaa tagtaacttg gcaagtacat tactattgga
5341 agtacgccag ggtacattgg cagtactccc attgacgtca atggcggtaa atggcccgcg
5401 atggctgcca agtacatccc cattgacgtc aatggggagg ggcaatgacg caaatgggcg
5461 ttccattgac gtaaatgggc ggtaggcgtg cctaatggga ggtctatata agcaatgctc
5521 gtttagggaa c
//
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