| 质粒名称: | pPAmCherry1-C1 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 31929 |
| 存储实验室: | Vladislav Verkhusha |
|
载体骨架基本信息 |
|
| 载体名称: | pEGFP-C1 |
| 载体出品公司: | Clontech |
| 载体抗性: | 卡那霉素 |
| 空载体大小: | 4008 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: | -- |
| 插入大小: | 708 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: | NheI（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-C1 was a gift from Vladislav Verkhusha (Addgene plasmid # 31929 ; http://n2t.net/addgene:31929 ; RRID:Addgene_31929) |
| 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-C1载体 pPAmCherry1C1质粒基本信息
pPAmCherry1-C1质粒图谱 pPAmCherry1C1载体图谱和pPAmCherry1-C1载体序列 pPAmCherry1C1质粒序列多克隆位点信息
pPAmCherry1-C1质粒 pPAmCherry1C1载体简介
pPAmCherry1-C1质粒序列 pPAmCherry1C1载体序列
LOCUS pPAmCherry1-C1 4716 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pPAmCherry1-C1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4716)
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 4716)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.biofeng.com/
FEATURES Location/Qualifiers
source 1..4716
/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"
regulatory 601..610
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 607..1314
/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(752..770)
/label=mCherry-R
/note="mCherry, reverse primer"
misc_feature 1315..1380
/label=MCS
/note="multiple cloning site"
polyA_signal 1504..1625
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1541..1560)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 1595..1614
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(1632..2087)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(1769..1790)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 1981..2000
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 2114..2218
/gene="bla"
/label=AmpR promoter
promoter 2220..2577
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(2243..2263)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 2428..2563
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2490..2509
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 2612..3406
/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(2666..2685)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3276..3295
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(3594..3613)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 3638..3685
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 4014..4602
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4503..4522
/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 gctagcggtc
601 gccaccatgg tgagcaaggg cgaggaggat aacatggcca tcattaagga gttcatgcgc
661 ttcaaggtgc acatggaggg gtccgtgaac ggccacgtgt tcgagatcga gggcgagggc
721 gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa gggcggcccc
781 ctgcccttca cctgggacat cctgagccct cagttcatgt acggctccaa tgcctacgtg
841 aagcaccccg ccgacatccc cgactacttt aagctgtcct tccccgaggg cttcaagtgg
901 gagcgcgtga tgaaattcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg
961 caggacggcg agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc
1021 cccgtgatgc agaagaagac catgggctgg gaggccctct ccgagcggat gtaccccgag
1081 gacggcgccc tgaagggcga ggtcaagccc agagtgaagc tgaaggacgg cggccactac
1141 gacgctgagg tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac
1201 aacgtcaacc gcaagctgga catcaccagc cacaacgagg actacaccat cgtggagcag
1261 tacgagagag ccgagggccg ccactccacc ggcggcatgg acgagctgta caagtccgga
1321 ctcagatctc gagctcaagc ttcgaattct gcagtcgacg gtaccgcggg cccgggatcc
1381 accggatcta gataactgat cataatcagc cataccacat ttgtagaggt tttacttgct
1441 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt
1501 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
1561 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
1621 tcttaacgcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
1681 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
1741 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
1801 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
1861 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
1921 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
1981 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
2041 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt
2101 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
2161 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc
2221 tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
2281 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
2341 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
2401 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
2461 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
2521 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat
2581 cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
2641 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
2701 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
2761 gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct
2821 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
2881 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
2941 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
3001 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
3061 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
3121 gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
3181 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
3241 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
3301 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
3361 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
3421 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
3481 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
3541 cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca
3601 cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa
3661 acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct
3721 gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca
3781 ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc
3841 cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa
3901 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
3961 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
4021 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
4081 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
4141 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
4201 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
4261 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
4321 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
4381 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
4441 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
4501 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
4561 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
4621 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
4681 cctgattctg tggataaccg tattaccgcc atgcat
//
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