| 质粒名称: | pmiRFP720-N1 |
|---|---|
| 出品公司: | addgene |
| 目录编号: | 136560 |
| 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 |
| 空载体大小: | 4038 bp |
| 完整质粒大小: | 4989 bp |
| 载体修饰: | 人细胞色素c氧化酶亚基VIII（human cytochrome C oxidase subunit VIII） |
| 筛选标记: | 新霉素（G418） |
| 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: | miRFP720 |
| 物种来源: | 光合细菌 Rhodopseudomonas palustris |
| 插入大小: | 951 bp |
| 基因突变: | T2A, S13L, A92T, V104T, V114I, E161K, Y193K, F198Y, D202T, I203V, Y258F, A283V, K288T, N290Y relative to RpBphP2 |
| GenBank编号 | JN247409 |
| 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: | NheI(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: | pmiRFP720-N1 was a gift from Vladislav Verkhusha (Addgene plasmid # 136560 ; http://n2t.net/addgene:136560 ; RRID:Addgene_136560) |
| 参考文献部分: | A set of monomeric near-infrared fluorescent proteins for multicolor imaging across scales. Matlashov ME, Shcherbakova DM, Alvelid J, Baloban M, Pennacchietti F, Shemetov AA, Testa I, Verkhusha VV. Nat Commun. 2020 Jan 13;11(1):239. doi: 10.1038/s41467-019-13897-6. 10.1038/s41467-019-13897-6 PubMed 31932632 |
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 pmiRFP720-N1 4887 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION A cytoplasmic expression of a monomeric phytochrome-based
near-infrared fluorescent protein miRFP720.
ACCESSION .
VERSION .
KEYWORDS pmiRFP720-N1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4887)
AUTHORS Matlashov ME, Shcherbakova DM, Alvelid J, Baloban M, Pennacchietti
F, Shemetov AA, Testa I, Verkhusha VV
TITLE A set of monomeric near-infrared fluorescent proteins for multicolor
imaging across scales.
JOURNAL Nat Commun. 2020 Jan 13;11(1):239. doi: 10.1038/s41467-019-13897-6.
PUBMED 31932632
REFERENCE 2 (bases 1 to 4887)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..4887
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 33..336
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 337..540
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 491..511
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
regulatory 569..578
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
polyA_signal 1647..1768
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1684..1703)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 1738..1757
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(1775..2230)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(1912..1933)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2124..2143
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 2257..2361
/gene="bla"
/label=AmpR promoter
promoter 2363..2720
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(2386..2406)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 2571..2706
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2633..2652
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 2755..3549
/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(2809..2828)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3419..3438
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(3737..3756)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 3781..3828
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 4157..4745
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4646..4665
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
61 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
121 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
181 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
241 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
301 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc
361 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
421 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc
481 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct
541 ggtttagtga accgtcagat ccgctagcgc caccatggcg gaaggctccg tcgccaggca
601 gcctgacctc ttgacctgcg acgatgagcc gatccatatc cccggtgcca tccaaccgca
661 tggactgctg ctcgccctcg ccgccgacat gacgatcgtt gccggcagcg acaaccttcc
721 cgaactcacc ggactggcga tcggcgccct gatcggccgc tctgcggccg atgtcttcga
781 ctcggagacg cacaaccgtc tgacgatcgc cttggccgag cccggggcgg ccgtcggagc
841 accgatcact gtcggcttca cgatgcgaaa ggacgcaggc ttcatcggct cctggcatcg
901 ccatgatcag ctcatcttcc tggagctgga gcctccccag cgggacgtcg ccgagccgca
961 ggcgttcttc cgccgcacca acagcgccat ccgccgcctg caggccgccg aaaccttgga
1021 aagcgcctgc gccgccgcgg cgcaagaggt gcggaagatt accggcttcg atcgggtgat
1081 gatctatcgc ttcgcctccg acttcagcgg gtccgtgatc gcagaggatc ggtgcgccga
1141 ggtcgagtca aaactaggcc tgcactatcc tgcctcattc atcccggcgc aggcccgtcg
1201 gctctatacc atcaacccgg tacggatcat tcccgacatc aattatcggc cggtgccggt
1261 caccccagac ctcaatccgg tcaccgggcg gccgattgat cttagcttcg ccatcctgcg
1321 cagcgtctcg cccaaccatc tggagttcat gcgcaacata ggcatgcacg gcacgatgtc
1381 gatctcgatt ttgcgcggcg agcgactgtg gggattgatc gtttgccatc accgaacgcc
1441 gtactacgtc gatctcgatg gccgccaagc ctgcaagagg gtcgccgaga ggctggccac
1501 tcagatcggc gtgatggaag agtgagcggc cgcgactcta gatcataatc agccatacca
1561 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
1621 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
1681 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
1741 gtttgtccaa actcatcaat gtatcttaag gcgtaaattg taagcgttaa tattttgtta
1801 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
1861 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
1921 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
1981 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
2041 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
2101 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
2161 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
2221 cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
2281 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
2341 taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt
2401 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
2461 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
2521 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
2581 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
2641 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
2701 gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt
2761 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
2821 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
2881 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac
2941 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
3001 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
3061 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
3121 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
3181 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
3241 caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag
3301 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
3361 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
3421 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
3481 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
3541 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
3601 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
3661 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
3721 ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga
3781 cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg
3841 ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg
3901 cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg
3961 cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga
4021 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
4081 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
4141 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
4201 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
4261 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
4321 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
4381 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
4441 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
4501 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
4561 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
4621 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
4681 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
4741 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
4801 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcatt
4861 agttattaat agtaatcaat tacgggg
//
