| 质粒名称: | pmiRFP670-N1 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 79987 |
| 存储实验室: | Vladislav Verkhusha |
|
载体骨架基本信息 |
|
| 载体名称: | pN1 |
| 载体出品公司: | Clontech |
| 载体抗性: | 卡那霉素 |
| 空载体大小: | 4012 bp |
| 完整质粒大小: | 4960 bp |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 新霉素（G418） |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | miRFP670 |
| 物种来源: | 光合细菌 Rhodopseudomonas palustris |
| 插入大小: | 948 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | CMV |
| 标签/融合蛋白: | 人细胞色素c氧化酶亚基VIII（N端） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | AgeI(未破坏) |
| 3' 克隆位点: | NotI (未破坏) |
| 5' 测序引物 | CMV-F |
| 3' 测序引物 | SV40pA-R |
|
文献引用方法 |
|
| 材料和方法部分: | pmiRFP670-N1 was a gift from Vladislav Verkhusha (Addgene plasmid # 79987 ; http://n2t.net/addgene:79987 ; RRID:Addgene_79987) |
| 参考文献部分: | Bright monomeric near-infrared fluorescent proteins as tags and biosensors for multiscale imaging. Shcherbakova DM, Baloban M, Emelyanov AV, Brenowitz M, Guo P, Verkhusha VV. Nat Commun. 2016 Aug 19;7:12405. doi: 10.1038/ncomms12405. 10.1038/ncomms12405 PubMed 27539380 |
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载体基本信息
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载体简介
载体序列
LOCUS pmiRFP670-N1 4960 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Monomeric near-infrared fluorescent protein miRFP670 in mammalian
plasmid.
ACCESSION .
VERSION .
KEYWORDS pmiRFP670-N1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4960)
AUTHORS Shcherbakova DM, Baloban M, Emelyanov AV, Brenowitz M, Guo P,
Verkhusha VV
TITLE Bright monomeric near-infrared fluorescent proteins as tags and
biosensors for multiscale imaging.
JOURNAL Nat Commun. 2016 Aug 19;7:12405. doi: 10.1038/ncomms12405.
PUBMED 27539380
REFERENCE 2 (bases 1 to 4960)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..4960
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 106..409
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 410..613
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 564..584
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
misc_feature 636..716
/label=MCS
/note="multiple cloning site"
regulatory 718..727
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 724..1671
/codon_start=1
/product="bright monomeric near-infrared fluorescent
protein engineered from a bacterial phytochrome, with an
emission maximum of 670 nm (Shcherbakova et al., 2016)"
/label=miRFP670
/label=ORF frame 1
/translation="MVAGHASGSPAFGTASHSNCEHEEIHLAGSIQPHGALLVVSEHDH
RVIQASANAAEFLNLGSVLGVPLAEIDGDLLIKILPHLDPTAEGMPVAVRCRIGNPSTE
YCGLMHRPPEGGLIIELERAGPSIDLSGTLAPALERIRTAGSLRALCDDTVLLFQQCTG
YDRVMVYRFDEQGHGLVFSECHVPGLESYFGNRYPSSTVPQMARQLYVRQRVRVLVDVT
YQPVPLEPRLSPLTGRDLDMSGCFLRSMSPCHLQFLKDMGVRATLAVSLVVGGKLWGLV
VCHHYLPRFIRFELRAICKRLAERIATRITALES"
polyA_signal 1793..1914
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1830..1849)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 1884..1903
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(1921..2376)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2058..2079)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2270..2289
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 2403..2507
/gene="bla"
/label=AmpR promoter
promoter 2509..2866
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(2532..2552)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 2717..2852
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2779..2798
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 2901..3695
/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(2955..2974)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3565..3584
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(3883..3902)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 3927..3974
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 4303..4891
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4792..4811
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 gcgttatccc ctgattctgt ggataaccgt attaccgcca tgcattagtt attaatagta
61 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
121 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
181 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
241 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
301 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
361 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
421 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
481 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
541 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
601 tataagcaga gctggtttag tgaaccgtca gatccgctag cgctaccgga ctcagatctc
661 gagctcaagc ttcgaattct gcagtcgacg gtaccgcggg cccgggatcc accggtcgcc
721 accatggtag caggtcatgc ctctggcagc cccgcattcg ggaccgcctc tcattcgaat
781 tgcgaacatg aagagatcca cctcgccggc tcgatccagc cgcatggcgc gcttctggtc
841 gtcagcgaac atgatcatcg cgtcatccag gccagcgcca acgccgcgga atttctgaat
901 ctcggaagcg tactcggcgt tccgctcgcc gagatcgacg gcgatctgtt gatcaagatc
961 ctgccgcatc tcgatcccac cgccgaaggc atgccggtcg cggtgcgctg ccggatcggc
1021 aatccctcta cggagtactg cggtctgatg catcggcctc cggaaggcgg gctgatcatc
1081 gaactcgaac gtgccggccc gtcgatcgat ctgtcaggca cgctggcgcc ggcgctggag
1141 cggatccgca cggcgggttc actgcgcgcg ctgtgcgatg acaccgtgct gctgtttcag
1201 cagtgcaccg gctacgaccg ggtgatggtg tatcgtttcg atgagcaagg ccacggcctg
1261 gtattctccg agtgccatgt gcctgggctc gaatcctatt tcggcaaccg ctatccgtcg
1321 tcgactgtcc cgcagatggc gcggcagctg tacgtgcggc agcgcgtccg cgtgctggtc
1381 gacgtcacct atcagccggt gccgctggag ccgcggctgt cgccgctgac cgggcgcgat
1441 ctcgacatgt cgggctgctt cctgcgctcg atgtcgccgt gccatctgca gttcctgaag
1501 gacatgggcg tgcgcgccac cctggcggtg tcgctggtgg tcggcggcaa gctgtggggc
1561 ctggttgtct gtcaccatta tctgccgcgc ttcatccgtt tcgagctgcg ggcgatctgc
1621 aaacggctcg ccgaaaggat cgcgacgcgg atcaccgcgc ttgagagcta agcggccgcg
1681 actctagatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct
1741 cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt
1801 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
1861 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaaggcgt
1921 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
1981 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
2041 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
2101 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
2161 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
2221 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
2281 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
2341 cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg
2401 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
2461 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa
2521 gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
2581 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
2641 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
2701 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
2761 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
2821 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga
2881 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
2941 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
3001 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
3061 ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg
3121 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
3181 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
3241 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
3301 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
3361 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
3421 tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
3481 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
3541 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
3601 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
3661 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
3721 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
3781 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
3841 tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac
3901 aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt
3961 tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc
4021 accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc
4081 aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc
4141 ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
4201 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
4261 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
4321 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
4381 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
4441 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
4501 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
4561 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
4621 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
4681 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
4741 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
4801 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
4861 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
4921 cctggccttt tgctggcctt ttgctcacat gttctttcct
//
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