| 质粒名称: | pPAmCherry1-N1 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 31928 |
| 存储实验室: | Vladislav Verkhusha |
|
载体骨架基本信息 |
|
| 载体名称: | pEGFP-N1 |
| 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: | -- |
| 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: | PAmCherry1,photoactivatable dark-to-red mCherry red fluorescent protein |
| 物种来源: | 光合细菌 Rhodopseudomonas palustris |
| 插入大小: | 711 bp |
| 基因突变: | E26V/A58T/K69N/L84F/N99K/S148L/I165V/Q167P/L169V/I203R compared to mCherry (but numbering is relative to EGFP). |
| GenBank编号 | -- |
| 启动子 | CMV |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | AgeI（销毁时未知） |
| 3' 克隆位点: | BsrGI（销毁时未知） |
| 5' 测序引物 | cggtaggcgtgtacggtgggag |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pPAmCherry1-N1 was a gift from Vladislav Verkhusha (Addgene plasmid # 31928 ; http://n2t.net/addgene:31928 ; RRID:Addgene_31928) |
| 参考文献部分: | Photoactivatable mCherry for high-resolution two-color fluorescence microscopy. Subach FV, Patterson GH, Manley S, Gillette JM, Lippincott-Schwartz J, Verkhusha VV. Nat Methods. 2009 Feb;6(2):153-9. Epub 2009 Jan 25. 10.1038/nmeth.1298 PubMed 19169259 |
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pPAmCherry1-N1载体 pPAmCherry1N1质粒基本信息
pPAmCherry1-N1质粒图谱 pPAmCherry1N1载体图谱和pPAmCherry1-N1载体序列 pPAmCherry1N1质粒序列多克隆位点信息
pPAmCherry1-N1质粒 pPAmCherry1N1载体简介
pPAmCherry1-N1质粒序列 pPAmCherry1N1载体序列
LOCUS pPAmCherry1-N1 4723 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pPAmCherry1-N1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4723)
AUTHORS Subach FV, Patterson GH, Manley S, Gillette JM, Lippincott-Schwartz
J, Verkhusha VV
TITLE Photoactivatable mCherry for high-resolution two-color fluorescence
microscopy.
JOURNAL Nat Methods. 2009 Feb;6(2):153-9. Epub 2009 Jan 25.
PUBMED 19169259
REFERENCE 2 (bases 1 to 4723)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..4723
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 61..364
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 365..568
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 519..539
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
misc_feature 591..671
/label=MCS
/note="multiple cloning site"
regulatory 673..682
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 679..1389
/codon_start=1
/product="photoactivatable monomeric derivative of DsRed
fluorescent protein (Subach et al., 2009)"
/label=PAmCherry1
/note="mammalian codon-optimized"
/translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHVFEIEGEGEGRPYEG
TQTAKLKVTKGGPLPFTWDILSPQFMYGSNAYVKHPADIPDYFKLSFPEGFKWERVMKF
EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEALSERMYPEDGALK
GEVKPRVKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNRKLDITSHNEDYTIVEQYERA
EGRHSTGGMDELYK"
primer_bind complement(824..842)
/label=mCherry-R
/note="mCherry, reverse primer"
polyA_signal 1511..1632
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1548..1567)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 1602..1621
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(1639..2094)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(1776..1797)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 1988..2007
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 2121..2225
/gene="bla"
/label=AmpR promoter
promoter 2227..2584
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(2250..2270)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 2435..2570
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2497..2516
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 2619..3413
/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(2673..2692)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3283..3302
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(3601..3620)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 3645..3692
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 4021..4609
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4510..4529
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
601 ccggactcag atctcgagct caagcttcga attctgcagt cgacggtacc gcgggcccgg
661 gatccaccgg tcgccaccat ggtgagcaag ggcgaggagg ataacatggc catcattaag
721 gagttcatgc gcttcaaggt gcacatggag gggtccgtga acggccacgt gttcgagatc
781 gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc
841 aagggcggcc ccctgccctt cacctgggac atcctgagcc ctcagttcat gtacggctcc
901 aatgcctacg tgaagcaccc cgccgacatc cccgactact ttaagctgtc cttccccgag
961 ggcttcaagt gggagcgcgt gatgaaattc gaggacggcg gcgtggtgac cgtgacccag
1021 gactcctccc tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg caccaacttc
1081 ccctccgacg gccccgtgat gcagaagaag accatgggct gggaggccct ctccgagcgg
1141 atgtaccccg aggacggcgc cctgaagggc gaggtcaagc ccagagtgaa gctgaaggac
1201 ggcggccact acgacgctga ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg
1261 cccggcgcct acaacgtcaa ccgcaagctg gacatcacca gccacaacga ggactacacc
1321 atcgtggagc agtacgagag agccgagggc cgccactcca ccggcggcat ggacgagctg
1381 tacaagtaag cggccgcgac tctagatcat aatcagccat accacatttg tagaggtttt
1441 acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat
1501 tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
1561 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
1621 caatgtatct taaggcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
1681 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
1741 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
1801 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
1861 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
1921 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
1981 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
2041 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg
2101 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
2161 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
2221 agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc
2281 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
2341 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
2401 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
2461 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
2521 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
2581 caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
2641 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
2701 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
2761 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat
2821 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
2881 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
2941 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
3001 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
3061 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
3121 ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc
3181 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
3241 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
3301 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
3361 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
3421 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
3481 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
3541 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac
3601 tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca
3661 gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg
3721 gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt
3781 ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc
3841 ggcaggccct gccatagcct caggttactc atatatactt tagattgatt taaaacttca
3901 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
3961 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
4021 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
4081 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
4141 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
4201 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
4261 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
4321 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
4381 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
4441 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
4501 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
4561 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
4621 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
4681 gttatcccct gattctgtgg ataaccgtat taccgccatg cat
//
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