| 质粒名称: | Lamp1-RFP |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 1817 |
| 存储实验室: | Walther Mothes |
|
载体骨架基本信息 |
|
| 载体名称: | Modified Clontech Plasmid |
| 载体出品公司: | -- |
| 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（对质粒功能不重要） |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | RFP（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 Forward |
| 3' 测序引物 | DsRed1-N |
|
文献引用方法 |
|
| 材料和方法部分: | Lamp1-RFP was a gift from Walther Mothes (Addgene plasmid # 1817 ; http://n2t.net/addgene:1817 ; RRID:Addgene_1817) |
| 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
Lamp1RFP载体基本信息
Lamp1RFP载体图谱质粒图谱和多克隆位点信息
Lamp1RFP载体简介
Lamp1RFP载体序列
LOCUS Lamp1-RFP 5906 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Mammalian expression of Lamp1 fused to RFP. Used for localization to
the lysosome..
ACCESSION .
VERSION .
KEYWORDS Lamp1-RFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5906)
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 5906)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5906
/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..2559
/codon_start=1
/product="monomeric derivative of DsRed (Campbell et al.,
2002)"
/label=mRFP1
/note="mammalian codon-optimized"
/translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
TGA"
primer_bind complement(2015..2033)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind 2283..2302
/label=mCherry-F
/note="mCherry, forward primer"
primer_bind 2477..2500
/label=DsRed1-C
/note="DsRed1, forward primer"
polyA_signal 2694..2815
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2731..2750)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 2785..2804
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(2822..3277)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2959..2980)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 3171..3190
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 3304..3408
/gene="bla"
/label=AmpR promoter
promoter 3410..3767
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(3433..3453)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 3618..3753
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3680..3699
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3802..4596
/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(3856..3875)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 4466..4485
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(4784..4803)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 4828..4875
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 5204..5792
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5693..5712
/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 caccatggcc tcctccgagg acgtcatcaa ggagttcatg
1921 cgcttcaagg tgcgcatgga gggctccgtg aacggccacg agttcgagat cgagggcgag
1981 ggcgagggcc gcccctacga gggcacccag accgccaagc tgaaggtgac caagggcggc
2041 cccctgccct tcgcctggga catcctgtcc cctcagttcc agtacggctc caaggcctac
2101 gtgaagcacc ccgccgacat ccccgactac ttgaagctgt ccttccccga gggcttcaag
2161 tgggagcgcg tgatgaactt cgaggacggc ggcgtggtga ccgtgaccca ggactcctcc
2221 ctgcaggacg gcgagttcat ctacaaggtg aagctgcgcg gcaccaactt cccctccgac
2281 ggccccgtaa tgcagaagaa gaccatgggc tgggaggcct ccaccgagcg gatgtacccc
2341 gaggacggcg ccctgaaggg cgagatcaag atgaggctga agctgaagga cggcggccac
2401 tacgacgccg aggtcaagac cacctacatg gccaagaagc ccgtgcagct gcccggcgcc
2461 tacaagaccg acatcaagct ggacatcacc tcccacaacg aggactacac catcgtggaa
2521 cagtacgagc gcgccgaggg ccgccactcc accggcgcct tgtacaagta aagcggccgc
2581 gactctagat cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc
2641 tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt
2701 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
2761 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttaaggcg
2821 taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
2881 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
2941 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
3001 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
3061 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
3121 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
3181 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
3241 acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat
3301 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
3361 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa
3421 agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
3481 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
3541 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
3601 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
3661 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
3721 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag
3781 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
3841 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
3901 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
3961 tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg
4021 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
4081 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
4141 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
4201 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
4261 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
4321 ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
4381 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
4441 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
4501 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
4561 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
4621 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
4681 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
4741 atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga
4801 caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg
4861 ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc
4921 caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc
4981 caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag
5041 cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
5101 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
5161 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
5221 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
5281 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
5341 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
5401 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
5461 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
5521 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
5581 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
5641 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
5701 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
5761 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
5821 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
5881 tggataaccg tattaccgcc atgcat
//
