| 质粒名称: | DsRed-rab7 WT |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 12661 |
| 存储实验室: | Richard Pagano |
|
载体骨架基本信息 |
|
| 载体名称: | pDsRed-C1 |
| 载体出品公司: | Clontech |
| 载体抗性: | 卡那霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | 4700 bp |
| 载体修饰: | 人细胞色素c氧化酶亚基VIII（human cytochrome C oxidase subunit VIII） |
| 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: | RAB7 (Homo sapiens) |
| 物种来源: | -- |
| 插入大小: | 693 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | 人细胞色素c氧化酶亚基VIII（N端） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | AgeI(未破坏) |
| 3' 克隆位点: | NotI (未破坏) |
| 5' 测序引物 | CMV-F |
| 3' 测序引物 | SV40pA-R |
|
文献引用方法 |
|
| 材料和方法部分: | DsRed-rab7 WT was a gift from Richard Pagano (Addgene plasmid # 12661 ; http://n2t.net/addgene:12661 ; RRID:Addgene_12661) |
| 参考文献部分: | Rab proteins mediate Golgi transport of caveola-internalized glycosphingolipids and correct lipid trafficking in Niemann-Pick C cells. Choudhury A, Dominguez M, Puri V, Sharma DK, Narita K, Wheatley CL, Marks DL, Pagano RE. J Clin Invest. 2002 Jun . 109(12):1541-50. 10.1172/JCI15420 PubMed 12070301 |
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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 DsRed-rab7 WT 6060 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS DsRed-rab7 WT
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6060)
AUTHORS Choudhury A, Dominguez M, Puri V, Sharma DK, Narita K, Wheatley CL,
Marks DL, Pagano RE
TITLE Rab proteins mediate Golgi transport of caveola-internalized
glycosphingolipids and correct lipid trafficking in Niemann-Pick C
cells.
JOURNAL J Clin Invest. 2002 Jun . 109(12):1541-50.
PUBMED 12070301
REFERENCE 2 (bases 1 to 6060)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6060
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 68..371
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 372..575
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 526..546
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
regulatory 614..623
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 620..1297
/codon_start=1
/product="wild-type DsRed"
/label=DsRed1
/note="mammalian codon-optimized"
/translation="MVRSSKNVIKEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTV
KLKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
VVTVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
KALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERTEGRH
HLFL"
primer_bind complement(803..823)
/label=DsRed1-N
/note="DsRed1, reverse primer"
primer_bind 1021..1040
/label=mCherry-F
/note="mCherry, forward primer"
primer_bind 1215..1238
/label=DsRed1-C
/note="DsRed1, forward primer"
misc_feature 1301..1357
/label=MCS
/note="multiple cloning site"
CDS 1361..1387
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
CDS 1388..1414
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
CDS 2048..2074
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
CDS 2075..2101
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
polyA_signal 2855..2976
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2892..2911)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 2946..2965
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(2983..3438)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3120..3141)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 3332..3351
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 3465..3569
/gene="bla"
/label=AmpR promoter
promoter 3571..3928
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(3594..3614)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 3779..3914
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3841..3860
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3963..4757
/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(4017..4036)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 4627..4646
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(4945..4964)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 4989..5036
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 5365..5953
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5854..5873
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
61 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
121 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
181 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
241 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
301 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
361 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
421 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
481 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
541 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
601 agcgctaccg gtcgccacca tggtgcgctc ctccaagaac gtcatcaagg agttcatgcg
661 cttcaaggtg cgcatggagg gcaccgtgaa cggccacgag ttcgagatcg agggcgaggg
721 cgagggccgc ccctacgagg gccacaacac cgtgaagctg aaggtgacca agggcggccc
781 cctgcccttc gcctgggaca tcctgtcccc ccagttccag tacggctcca aggtgtacgt
841 gaagcacccc gccgacatcc ccgactacaa gaagctgtcc ttccccgagg gcttcaagtg
901 ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc gtgacccagg actcctccct
961 gcaggacggc tgcttcatct acaaggtgaa gttcatcggc gtgaacttcc cctccgacgg
1021 ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc accgagcgcc tgtacccccg
1081 cgacggcgtg ctgaagggcg agatccacaa ggccctgaag ctgaaggacg gcggccacta
1141 cctggtggag ttcaagtcca tctacatggc caagaagccc gtgcagctgc ccggctacta
1201 ctacgtggac tccaagctgg acatcacctc ccacaacgag gactacacca tcgtggagca
1261 gtacgagcgc accgagggcc gccaccacct gttcctgctc agatctcgag ctcaagcttc
1321 gaattctgca gtcgacggta ccgcgggccc gggatccatg tacccatacg atgttccgga
1381 ttacgcttac ccatacgatg ttccggatta cgctggtacc tctaggaaga aagtgttgct
1441 gaaggttatc atcctgggag attctggagt cgggaagaca tcactcatga accagtatgt
1501 gaataagaaa ttcagcaatc agtacaaagc cacaatagga gctgactttc tgaccaagga
1561 ggtgatggtg gatgacaggc tagtcacaat gcagatatgg gacacagcag gacaggaacg
1621 gttccagtct ctcggtgtgg ccttctacag aggtgcagac tgctgcgttc tggtatttga
1681 tgtgactgcc cccaacacat tcaaaaccct agatagctgg agagatgagt ttctcatcca
1741 ggccagtccc cgagatcctg aaaacttccc atttgttgtg ttgggaaaca agattgacct
1801 cgaaaacaga caagtggcca caaagcgggc acaggcctgg tgctacagca aaaacaacat
1861 tccctacttt gagaccagtg ccaaggaggc catcaacgtg gagcaggcgt tccagacgat
1921 tgcacggaat gcacttaagc aggaaacgga ggtggagctg tacaacgaat ttcctgaacc
1981 tatcaaactg gacaagaatg accgggccaa ggcctcggca gaaagctgca gttgctgagg
2041 atccatgtac ccatacgatg ttccggatta cgcttaccca tacgatgttc cggattacgc
2101 tggtacctct aggaagaaag tgttgctgaa ggttatcatc ctgggagatt ctggagtcgg
2161 gaagacatca ctcatgaacc agtatgtgaa taagaaattc agcaatcagt acaaagccac
2221 aataggagct gactttctga ccaaggaggt gatggtggat gacaggctag tcacaatgca
2281 gatatgggac acagcaggac aggaacggtt ccagtctctc ggtgtggcct tctacagagg
2341 tgcagactgc tgcgttctgg tatttgatgt gactgccccc aacacattca aaaccctaga
2401 tagctggaga gatgagtttc tcatccaggc cagtccccga gatcctgaaa acttcccatt
2461 tgttgtgttg ggaaacaaga ttgacctcga aaacagacaa gtggccacaa agcgggcaca
2521 ggcctggtgc tacagcaaaa acaacattcc ctactttgag accagtgcca aggaggccat
2581 caacgtggag caggcgttcc agacgattgc acggaatgca cttaagcagg aaacggaggt
2641 ggagctgtac aacgaatttc ctgaacctat caaactggac aagaatgacc gggccaaggc
2701 ctcggcagaa agctgcagtt gctgaggatc caccggatct agataactga tcataatcag
2761 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa
2821 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
2881 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
2941 tagttgtggt ttgtccaaac tcatcaatgt atcttaacgc gtaaattgta agcgttaata
3001 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
3061 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
3121 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
3181 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
3241 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
3301 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
3361 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
3421 cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
3481 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
3541 tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt
3601 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
3661 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
3721 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
3781 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
3841 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
3901 cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc
3961 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
4021 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
4081 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
4141 tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
4201 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
4261 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
4321 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
4381 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
4441 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg
4501 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
4561 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
4621 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
4681 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
4741 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
4801 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
4861 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
4921 cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg
4981 cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa
5041 acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg
5101 ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc
5161 agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat
5221 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
5281 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
5341 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
5401 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
5461 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
5521 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
5581 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
5641 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
5701 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
5761 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
5821 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
5881 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
5941 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
6001 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
//
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