| 质粒名称: | Lamp1-YFP;pEYFP-N1-Lamp1-r |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 1816 |
| 存储实验室: | Walther Mothes |
|
载体骨架基本信息 |
|
| 载体名称: | pEYFP-N1 |
| 载体出品公司: | BD Biosciences / Clontech |
| 载体抗性: | 卡那霉素 |
| 空载体大小: | 4700 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: | lysosome associated membrane protein 1,Lamp-1 |
| 物种来源: | R. norvegicus (rat) |
| 插入大小: | 1200 bp |
| 基因突变: | D50E (not important for function of plasmid) |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | YFP（主干上的C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: | BamHI(未破坏) |
| 5' 测序引物 | CMV-F |
| 3' 测序引物 | EGFP-N |
|
文献引用方法 |
|
| 材料和方法部分: | Lamp1-YFP was a gift from Walther Mothes (Addgene plasmid # 1816 ; http://n2t.net/addgene:1816 ; RRID:Addgene_1816) |
| 参考文献部分: | Visualization of retroviral replication in living cells reveals budding into multivesicular bodies. Sherer NM, Lehmann MJ, Jimenez-Soto LF, Ingmundson A, Horner SM, Cicchetti G, Allen PG, Pypaert M, Cunningham JM, Mothes W. Traffic 2003 Nov;4(11):785-801. 10.1034/j.1600-0854.2003.00135.x PubMed 14617360 |
买家导航
Lamp1YFP载体基本信息
Lamp1YFP载体图谱质粒图谱和多克隆位点信息
Lamp1YFP载体简介
Lamp1YFP载体序列
LOCUS Lamp1-YFP 5939 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Mammalian expression of Lamp1 fused to YFP. Used for localization to
the lysosome..
ACCESSION .
VERSION .
KEYWORDS Lamp1-YFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5939)
AUTHORS Sherer NM, Lehmann MJ, Jimenez-Soto LF, Ingmundson A, Horner SM,
Cicchetti G, Allen PG, Pypaert M, Cunningham JM, Mothes W
TITLE Visualization of retroviral replication in living cells reveals
budding into multivesicular bodies.
JOURNAL Traffic 2003 Nov;4(11):785-801.
PUBMED 14617360
REFERENCE 2 (bases 1 to 5939)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5939
/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 637..646
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
regulatory 641..650
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 643..1863
/codon_start=1
/product="rat lysosome-associated membrane glycoprotein 1"
/label=LAMP1
/translation="MAAPGARRPLLLLLLAGLAHSAPALFEVKDNNGTACIMASFSASF
LTTYEAGHVSKVSNMTLPASAEVLKNSSSCGEKNASEPTLAITFGEGYLLKLTFTKNTT
RYSVQHMYFTYNLSDTQFFPNASSKGPDTVDSTTDIKADINKTYRCVSDIRVYMKNVTI
VLWDATIQAYLPSSNFSKEETRCPQDQPSPTTGPPSPSPPLVPTNPSVSKYNVTGDNGT
CLLASMALQLNITYMKKDNTTVTRAFNINPSDKYSGTCGAQLVTLKVGNKSRVLELQFG
MNATSSLFFLQGVQLNMTLPDAIEPTFSTSNYSLKALQASVGNSYKCNSEEHIFVSKAL
ALNVFSVQVQAFRVESDRFGSVEECVQDGNNMLIPIAVGGALAGLVLIVLIAYLIGRKR
SHAGYQTI"
regulatory 1879..1888
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 1885..2604
/codon_start=1
/product="enhanced YFP"
/label=EYFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(1930..1951)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(2191..2210)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 2538..2559
/label=EGFP-C
/note="EGFP, forward primer"
polyA_signal 2727..2848
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2764..2783)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 2818..2837
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(2855..3310)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2992..3013)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 3204..3223
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 3337..3441
/gene="bla"
/label=AmpR promoter
promoter 3443..3800
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(3466..3486)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 3651..3786
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3713..3732
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3835..4629
/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(3889..3908)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 4499..4518
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(4817..4836)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 4861..4908
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 5237..5825
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5726..5745
/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 attctcgcca ccatggcggc cccgggcgcc
661 cggcggccgc tgctcctgtt gctgctggca ggccttgcac acagcgcccc agcactgttc
721 gaggtgaaag acaacaacgg cacagcgtgt ataatggcca gcttctctgc ctcctttctg
781 accacctatg aggctggaca tgtttctaag gtctcgaata tgaccctgcc agcctctgca
841 gaagtcctga agaatagcag ctcttgtggt gaaaagaatg cttctgagcc caccctcgca
901 atcacctttg gagaaggata tttactgaaa ctcaccttca caaaaaacac aacacgttac
961 agtgtccagc acatgtattt cacatataac ctgtcagaca cacaattctt tcccaatgcc
1021 agctccaaag ggcccgacac tgtggattcc acaactgaca tcaaggcaga catcaacaaa
1081 acataccgat gtgtcagcga catcagggtc tacatgaaga atgtgaccat tgtgctctgg
1141 gacgctacta tccaggccta cctgccgagt agcaacttca gcaaggaaga gacacgctgc
1201 ccacaggatc aaccttcccc aactactggg ccacccagcc cctcaccacc actcgtgccc
1261 acaaacccca gtgtgtccaa gtacaatgtg actggtgaca atggaacctg cctgctggcc
1321 tctatggcac tgcaactcaa catcacctac atgaagaagg acaacacgac tgtgaccaga
1381 gcattcaaca tcaacccaag tgacaaatat agtgggactt gcggtgccca gttggtgacc
1441 ctgaaggtgg ggaacaagag cagagtcctg gagctgcagt ttgggatgaa tgccacttct
1501 agcctgtttt tcctgcaagg agttcagttg aacatgactc ttcctgatgc catagagccc
1561 acgttcagca cctccaacta ttccctgaaa gctcttcagg ccagtgtcgg caactcatac
1621 aagtgcaaca gtgaggagca catctttgtc agcaaggcgc tcgccctcaa tgtcttcagc
1681 gtgcaagtcc aggctttcag ggtagaaagt gacaggtttg ggtctgtgga agagtgtgta
1741 caggacggta acaacatgct gatccccatt gctgtgggcg gggccctggc agggctggtc
1801 ctcatcgtcc tcatcgccta cctcatcggc aggaagagga gtcacgcggg ctatcagacc
1861 atccgggatc caccggtcgc caccatggtg agcaagggcg aggagctgtt caccggggtg
1921 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
1981 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
2041 aagctgcccg tgccctggcc caccctcgtg accaccttcg gctacggcct gcagtgcttc
2101 gcccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
2161 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
2221 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
2281 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
2341 atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc
2401 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
2461 cccgtgctgc tgcccgacaa ccactacctg agctaccagt ccgccctgag caaagacccc
2521 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
2581 ggcatggacg agctgtacaa gtaaagcggc cgcgactcta gatcataatc agccatacca
2641 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
2701 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
2761 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
2821 gtttgtccaa actcatcaat gtatcttaag gcgtaaattg taagcgttaa tattttgtta
2881 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
2941 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
3001 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
3061 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
3121 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
3181 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
3241 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
3301 cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
3361 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
3421 taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt
3481 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
3541 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
3601 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
3661 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
3721 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
3781 gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt
3841 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
3901 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
3961 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac
4021 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
4081 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
4141 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
4201 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
4261 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
4321 caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag
4381 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
4441 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
4501 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
4561 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
4621 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
4681 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
4741 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
4801 ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga
4861 cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg
4921 ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg
4981 cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg
5041 cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga
5101 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
5161 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
5221 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
5281 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
5341 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
5401 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
5461 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
5521 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
5581 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
5641 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
5701 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
5761 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
5821 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
5881 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcat
//
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