| 质粒名称: | pmCherry-C1 mCherry-NLS |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 58476 |
| 存储实验室: | Dyche Mullins |
|
载体骨架基本信息 |
|
| 载体名称: | pmCherry-C1 |
| 载体出品公司: | Clontech |
| 载体抗性: | 卡那霉素 |
| 空载体大小: | 4700 bp |
| 完整质粒大小: | 4788 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: | 3X NLS with 3X stop codons |
| 物种来源: | SV40 virus |
| 插入大小: | 81 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | CMV |
| 标签/融合蛋白: | mCherry（插入件上的C端子） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | HindIII(未破坏) |
| 3' 克隆位点: | KpnI(未破坏) |
| 5' 测序引物 | mCherry Forward |
| 3' 测序引物 | SV40 polyA Reverse |
|
文献引用方法 |
|
| 材料和方法部分: | pmCherry-C1 mCherry-NLS was a gift from Dyche Mullins (Addgene plasmid # 58476 ; http://n2t.net/addgene:58476 ; RRID:Addgene_58476) |
| 参考文献部分: | DNA damage induces nuclear actin filament assembly by formin-2 and Spire-1/2 that promotes efficient DNA repair. Belin BJ, Lee T, Mullins RD. Elife. 2015 Aug 19;4. doi: 10.7554/eLife.07735. 10.7554/eLife.07735 PubMed 26287480 |
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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 pmCherry-C1 mCherry-NLS 4773 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Expresses a nuclear-targeted mCherry control construct, on a CMV
reporter.
ACCESSION .
VERSION .
KEYWORDS pmCherry-C1 mCherry-NLS
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4773)
AUTHORS Belin BJ, Lee T, Mullins RD
TITLE DNA damage induces nuclear actin filament assembly by formin-2 and
Spire-1/2 that promotes efficient DNA repair.
JOURNAL Elife. 2015 Aug 19;4. doi: 10.7554/eLife.07735.
PUBMED 26287480
REFERENCE 2 (bases 1 to 4773)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..4773
/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"
CDS 607..1314
/codon_start=1
/product="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
/label=mCherry
/note="mammalian codon-optimized"
/translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDTTSHNEDYTIVEQYERA
EGRHSTGGMDELYK"
primer_bind complement(752..770)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind 1020..1039
/label=mCherry-F
/note="mCherry, forward primer"
CDS 1342..1362
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 1366..1386
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
polyA_signal 1561..1682
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1598..1617)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 1652..1671
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(1689..2144)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(1826..1847)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2038..2057
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 2171..2275
/gene="bla"
/label=AmpR promoter
promoter 2277..2634
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(2300..2320)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 2485..2620
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2547..2566
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 2669..3463
/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(2723..2742)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3333..3352
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(3651..3670)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 3695..3742
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 4071..4659
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4560..4579
/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 ccggttatgg tgagcaaggg cgaggaggat aacatggcca tcatcaagga gttcatgcgc
661 ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc
721 gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc
781 ctgcccttcg cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg
841 aagcaccccg ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg
901 gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg
961 caggacggcg agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc
1021 cccgtaatgc agaagaagac catgggctgg gaggcctcct ccgagcggat gtaccccgag
1081 gacggcgccc tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac
1141 gacgctgagg tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac
1201 aacgtcaaca tcaagttgga caccacctcc cacaacgagg actacaccat cgtggaacag
1261 tacgaacgcg ccgagggccg ccactccacc ggcggcatgg acgagctgta caagagatct
1321 cgagctcaag cttccatgga tccaaaaaag aagagaaagg tagatccaaa aaagaagaga
1381 aaggtagatc caaaaaagaa gagaaagtga taatagggta ccgcgggccc gggatccacc
1441 ggatctagat aactgatcat aatcagccat accacatttg tagaggtttt acttgcttta
1501 aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt
1561 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
1621 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
1681 taacgcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
1741 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
1801 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
1861 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
1921 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
1981 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
2041 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
2101 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg
2161 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
2221 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga
2281 ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
2341 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
2401 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
2461 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
2521 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
2581 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga
2641 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
2701 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
2761 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
2821 cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc
2881 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
2941 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
3001 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
3061 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
3121 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
3181 cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
3241 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
3301 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
3361 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
3421 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
3481 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
3541 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
3601 cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg
3661 aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg
3721 cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc
3781 gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc
3841 caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct
3901 gccatagcct caggttactc atatatactt tagattgatt taaaacttca tttttaattt
3961 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
4021 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
4081 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
4141 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
4201 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
4261 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
4321 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
4381 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
4441 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
4501 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
4561 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
4621 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
4681 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
4741 gattctgtgg ataaccgtat taccgccatg cat
//
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