| 质粒名称: | pmCherry-C1-mTrim21 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 105516 |
| 存储实验室: | Melina Schuh |
|
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: | 4694 bp |
| 完整质粒大小: | 6083 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: | TRIM21,Ro52,Ssa1 |
| 物种来源: | M. musculus (mouse) |
| 插入大小: | 1389 bp |
| 基因突变: | -- |
| GenBank编号 | NM_001082552.2 |
| 启动子 | CMV |
| 标签/融合蛋白: | mCherry（主干上的N终端） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | EcoRI(未破坏) |
| 3' 克隆位点: | XbaI (未破坏) |
| 5' 测序引物 | CCCCGTAATGCAGAAGAAGA |
| 3' 测序引物 | GAAATTTGTGATGCTATTGC |
|
文献引用方法 |
|
| 材料和方法部分: | pmCherry-C1-mTrim21 was a gift from Melina Schuh (Addgene plasmid # 105516 ; http://n2t.net/addgene:105516 ; RRID:Addgene_105516) |
| 参考文献部分: | A Method for the Acute and Rapid Degradation of Endogenous Proteins. Clift D, McEwan WA, Labzin LI, Konieczny V, Mogessie B, James LC, Schuh M. Cell. 2017 Nov 13. pii: S0092-8674(17)31255-2. doi: 10.1016/j.cell.2017.10.033. 10.1016/j.cell.2017.10.033 PubMed 29153837 |
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 pmCherry-C1-mTrim21 6083 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Expression of m-Cherry-tagged wildtype mouse Trim21 in mammalian
cells.
ACCESSION .
VERSION .
KEYWORDS pmCherry-C1-mTrim21
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6083)
AUTHORS Clift D, McEwan WA, Labzin LI, Konieczny V, Mogessie B, James LC,
Schuh M
TITLE A Method for the Acute and Rapid Degradation of Endogenous Proteins.
JOURNAL Cell. 2017 Nov 13. pii: S0092-8674(17)31255-2. doi:
10.1016/j.cell.2017.10.033.
PUBMED 29153837
REFERENCE 2 (bases 1 to 6083)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6083
/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 607..616
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 613..1320
/codon_start=1
/product="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
/label=mCherry
/note="mammalian codon-optimized"
/translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
EGRHSTGGMDELYK"
primer_bind complement(758..776)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind 1026..1045
/label=mCherry-F
/note="mCherry, forward primer"
polyA_signal 2871..2992
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2908..2927)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 2962..2981
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(2999..3454)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3136..3157)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 3348..3367
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 3481..3585
/gene="bla"
/label=AmpR promoter
promoter 3587..3944
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(3610..3630)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 3795..3930
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3857..3876
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3979..4773
/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(4033..4052)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 4643..4662
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(4961..4980)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 5005..5052
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 5381..5969
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5870..5889
/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 ccggtcgcca ccatggtgag caagggcgag gaggataaca tggccatcat caaggagttc
661 atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc
721 gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt
781 ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc
841 tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc
901 aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc
961 tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc
1021 gacggccccg taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac
1081 cccgaggacg gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc
1141 cactacgacg ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc
1201 gcctacaacg tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg
1261 gaacagtacg aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag
1321 tccggactca gatctcgagc tcaagcttcg aattcaatgt ctctggaaaa gatgtgggag
1381 gaggtcacct gttctatctg cctggatccc atggtggagc ctatgagtat cgaatgtggc
1441 cattgctttt gcaaggaatg catttttgaa gttgggaaga atgggggcag ttcatgtccc
1501 gagtgccggc aacagtttct gctccgaaac ctcaggccca atagacatat agccaacatg
1561 gtggaaaacc ttaaacagat agcccagaat accaagaaga gtacccagga aacgcactgc
1621 atgaagcatg gagagaagct tcacctattc tgtgaggaag atgggcaggc cctttgctgg
1681 gtgtgtgccc agtctgggaa acaccgggac cacaccaggg tccctattga agaggctgct
1741 aaggtatacc aggagaagat ccacgtggct ttagaaaaac tgagaaaggg gaaagagttg
1801 gccgagaaga tggaaatgga tctcacgatg caaagaacag actggaagag gaacattgac
1861 acccagaagt cgaggattca cgcagagttc gcacttcaga atagcttgct ggctcaggag
1921 gagcagaggc agctgcagag gctggagaag gatcaaaggg agtacctgag actcctgggg
1981 aagaaggagg ctgagctggc tgagaagaac caggccctgc aggagctgat ctcagagctg
2041 gagaggagga ttcgtggttc agagctggag ctactgcagg aggtgaggat catcctggaa
2101 aggagtggat cctggaacct ggacacgtta gatattgacg ccccagacct aacaagcaca
2161 tgccctgtgc cagggcggaa gaagatgctg aggacgtgtt gggttcatat tactctggat
2221 cgcaacaccg ccaactcatg gctcatcatc tcaaaggatc ggagacaagt gaggatggga
2281 gacacccatc agaacgtgtc tgacaataag gagaggttta gtaattaccc catggtgcta
2341 ggtgcccaga gattctcctc tgggaagatg tactgggagg tagatgtgac tcaaaaggag
2401 gcctgggatc tgggggtttg cagagattct gttcagagga aagggcagtt ttcactcagt
2461 cccgagaatg gcttctggac catttggtta tggcaagaca gctatgaggc tggtaccagt
2521 cctcagacca ccctccacat tcaagtacct ccatgccaaa ttgggatctt tgtggactat
2581 gaggctggcg ttgtctcctt ctacaacata actgaccatg gctccctcat ttacaccttc
2641 tcggagtgtg tttttgctgg acctctgcga cctttcttca atgttggttt caattatagt
2701 gggggaaatg cagcgcctct aaagctctgt ccactaaaga tgtgagcggc cgctctagat
2761 aactgatcat aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc
2821 cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta
2881 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
2941 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct taacgcgtaa
3001 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
3061 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
3121 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
3181 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
3241 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
3301 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
3361 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
3421 cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg
3481 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
3541 caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga
3601 accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
3661 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
3721 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
3781 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
3841 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
3901 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca
3961 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
4021 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
4081 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
4141 ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc
4201 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
4261 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
4321 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
4381 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
4441 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
4501 aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
4561 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
4621 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
4681 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
4741 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
4801 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
4861 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
4921 tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa
4981 taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg
5041 ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac
5101 cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa
5161 gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct
5221 caggttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
5281 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
5341 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
5401 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
5461 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
5521 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
5581 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
5641 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
5701 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
5761 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
5821 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
5881 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
5941 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
6001 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
6061 ataaccgtat taccgccatg cat
//
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