| 质粒名称: | pEGFP-rNFM |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 32907 |
| 存储实验室: | Anthony Brown |
|
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: | 4134 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: | EGFP-rat NFM fusion,NFM,neurofilament protein M |
| 物种来源: | R. norvegicus (rat) |
| 插入大小: | 3330 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | CMV |
| 标签/融合蛋白: | EGFP（插入端N端） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | ApaI(未破坏) |
| 3' 克隆位点: | ApaI（克隆过程中销毁） |
| 5' 测序引物 | -- |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pEGFP-rNFM was a gift from Anthony Brown (Addgene plasmid # 32907 ; http://n2t.net/addgene:32907 ; RRID:Addgene_32907) |
| 参考文献部分: | Rapid movement of axonal neurofilaments interrupted by prolonged pauses. Wang L, Ho CL, Sun D, Liem RK, Brown A. Nat Cell Biol. 2000 Mar;2(3):137-41. 10.1038/35004008 PubMed 10707083 |
买家导航
pEGFP-rNFM载体 pEGFPrNFM质粒基本信息
pEGFP-rNFM质粒图谱 pEGFPrNFM载体图谱和pEGFP-rNFM载体序列 pEGFPrNFM质粒序列多克隆位点信息
pEGFP-rNFM质粒 pEGFPrNFM载体简介
pEGFP-rNFM质粒序列 pEGFPrNFM载体序列
LOCUS pEGFP-rNFM 7464 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Mammalian expression construct driving the expression of a
fluorescent fusion protein consisting of EGFP linked to the
N-terminus of rat neurofilament protein M.
ACCESSION .
VERSION .
KEYWORDS pEGFP-rNFM
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7464)
AUTHORS Wang L, Ho CL, Sun D, Liem RK, Brown A
TITLE Rapid movement of axonal neurofilaments interrupted by prolonged
pauses.
JOURNAL Nat Cell Biol. 2000 Mar;2(3):137-41.
PUBMED 10707083
REFERENCE 2 (bases 1 to 7464)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7464
/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..1329
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(658..679)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(919..938)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1266..1287
/label=EGFP-C
/note="EGFP, forward primer"
RBS 2854..2865
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
polyA_signal 4252..4373
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4289..4308)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4343..4362
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(4380..4835)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4517..4538)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 4729..4748
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 4862..4966
/gene="bla"
/label=AmpR promoter
promoter 4968..5325
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(4991..5011)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 5176..5311
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 5238..5257
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 5360..6154
/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(5414..5433)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 6024..6043
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(6342..6361)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 6386..6433
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 6762..7350
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 7251..7270
/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 gagctgttca ccggggtggt gcccatcctg
661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
1321 ctgtacaagt ccggactcag atctcgagct caagcttcga attctgcagt cgacggtacc
1381 gcgggcccaa ttccggcctc caagatgagc tacacgctgg actcgctggg caacccgtcc
1441 gcctaccggc gggtcaccga gacccgctcc agcttcagtc gtgtgagcgg ttccccgtcc
1501 agcggcttcc gctcgcagtc ctggtcccgc ggctcgccca gcaccgtgtc ctcctcctac
1561 aagcgcagcg cgctcgcccc gcgcctcgcc tacagctcgg ctatgctcag ctcggccgag
1621 agcagcctcg acttcagcca gtcctcttcg ctgcttaacg gcggctccgg cggcgactac
1681 aagctgtccc gctccaacga gaaagagcag ctgcaggggc tgaacgaccg cttcgccggc
1741 tacatcgaga aagtgcacta cttggaacaa cagaacaagg agatcgaggc agagatccac
1801 gcgctgcggc agaagcaggc ctcgcacgcc cagctgggtg acgcttacga ccaggagatc
1861 cgagagctgc gcgccaccct ggagatggtg aatcacgaga aggctcaagt gcagctggac
1921 tctgatcact tggaggaaga catccaccgg ctcaaggagc gcttcgagga ggaggcgcgg
1981 ctgcgggacg acaccgaggc tgccatccgg gcgctgcgca aagacataga ggagtcgtcg
2041 atggttaagg tggagctgga caagaaggtg cagtcgctgc aggatgaggt ggccttcctg
2101 cggagcaatc acgaagagga ggtggccgac ctgctggccc agatccaggc gtcgcacatc
2161 accgtagagc gcaaagacta cctgaagaca gacatctcca cggcgctgaa agagatccgc
2221 tcccagctcg agtgtcactc cgaccagaac atgcaccagg ccgaagagtg gttcaaatgc
2281 cgctacgcca agctcaccga ggcggccgag cagaacaagg aggccatccg ctccgctaaa
2341 gaagagatcg ccgagtaccg gcgccagctg cagtccaaga gcattgagct cgagtcggtg
2401 cgaggcacta aggagtccct ggaacggcag ctcagcgaca tcgaggagcg ccacaaccac
2461 gacctcagca gctaccagga caccatccag cagctggaaa atgagcttcg gggaacaaag
2521 tgggaaatgg ctcgtcattt gcgagaatac caggatctcc ttaacgtcaa gatggctctg
2581 gacatcgaga tcgccgcata taggaaacta ctggagggtg aagagaccag atttagcaca
2641 ttttcaggaa gcatcactgg gcctctgtac acacaccgac agccctcagt cacaatatcc
2701 agtaagattc agaagaccaa agtcgaggcc cccaagctca aggtccaaca caaatttgtg
2761 gaggagatca ttgaggagac taaagtggaa gatgagaagt cagaaatgga agacgccctc
2821 acagtcattg cagaggaatt ggcagcctct gccaaagagg agaaagaaga ggcagaagaa
2881 aaggaagagg aaccggaagt tgaaaagtct cccgtgaagt ctcctgaggc taaggaagag
2941 gaggaagggg aaaaggagga agaagaggaa ggccaagagg aagaagagga ggaagatgaa
3001 ggtgtcaagt cagaccaggc agaagaggga ggatctgaga aggaaggctc gagtgaaaag
3061 gatgaaggtg agcaagaaga agaaggggaa actgaggcag aaggtgaagg agaggaagca
3121 gaagctaagg aggaaaagaa aacagaggga aaggtcgagg aaatggctat caaggaggaa
3181 atcaaggtcg agaagcccga gaaagccaag tcccctgtgc caaaatcacc cgtggaagaa
3241 gtaaagccaa aaccagaagc caaagccgga aaggatgagc agaaggagga agagaaagtt
3301 gaggagaaga aggaggtagc caaggaatca cccaaggaag agaaggtgga gaaaaaggag
3361 gagaagccaa aagatgtccc agataaaaag aaggctgagt ccccagtgaa agaaaaggcc
3421 gtagaggaaa tgatcaccat tactaagtcg gtaaaggtga gcctggagaa agacaccaaa
3481 gaggagaagc ctcagcagca ggagaaggtg aaggagaagg cagaggagga ggggggtagt
3541 gaggaggaag tgggtgacaa aagcccgcaa gaatccaaga aggaagacat agctatcaat
3601 ggggaggtgg aaggaaaaga ggaggaggag caggaaactc aggagaaggg cagtgggcaa
3661 gaggaggaga aaggggtggt cactaatggc ttagatgtga gccctgcgga ggaaaagaaa
3721 ggggaggata gaagtgatga caaagtggtg gtgaccaaga aggtagaaaa aatcaccagc
3781 gagggaggcg atggtgctac caaatacatc accaaatctg ttactgtcac tcaaaaggtt
3841 gaagagcatg aggagacctt tgaggagaag ctggtgtcaa ctaaaaaggt agaaaaggtc
3901 acttcacatg ccatagtcaa ggaagtcacc cagggtgact aagatcggag tccattgcaa
3961 aaggttaagc catacaacaa tttcaaaatg catgtgattg acagcttcaa aacagaatgg
4021 gttctcccat gagggctcca gacattgtat tttcctttgt gcaatatgag ggaactgcat
4081 gcaagctcag ggtgcccccc tcctcagtcc ttgggggaat tgggatccac cggatctaga
4141 taactgatca taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc
4201 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt
4261 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
4321 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaacgcgta
4381 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
4441 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
4501 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
4561 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
4621 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
4681 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
4741 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
4801 acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt
4861 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
4921 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag
4981 aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
5041 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
5101 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
5161 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
5221 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
5281 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac
5341 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
5401 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
5461 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
5521 cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg
5581 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
5641 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
5701 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
5761 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
5821 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
5881 caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
5941 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
6001 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
6061 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
6121 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
6181 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
6241 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
6301 ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca
6361 ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt
6421 gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca
6481 ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca
6541 agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc
6601 tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
6661 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
6721 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
6781 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
6841 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
6901 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
6961 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
7021 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
7081 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
7141 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
7201 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
7261 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
7321 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
7381 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
7441 gataaccgta ttaccgccat gcat
//
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