| 质粒名称: | MLV Gag-RFP |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 1814 |
| 存储实验室: | Walther Mothes |
|
载体骨架基本信息 |
|
| 载体名称: | pmRFP-N1 |
| 空载体大小: | 4700 bp |
| 完整质粒大小: | 6832 bp |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 新霉素（G418） |
| 载体抗性: | Kanamycin |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | Gag |
| 物种来源: | MuLV |
| 插入大小: | 2100bp |
| 标签/融合蛋白: | RFP (C terminal on backbone) |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | BamHI(未破坏) |
| 3’克隆位点: | AgeI(未破坏) |
| 5’测序引物 | CMV-F |
|
文献引用方法 |
|
| 材料和方法部分: |
MLV Gag-RFP was a gift from Walther Mothes (Addgene plasmid # 1814 ; http://n2t.net/addgene:1814 ; RRID:Addgene_1814)
|
| 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
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 MLV Gag-RFP
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS MLV Gag-RFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6832)
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 6832)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..6832
/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"
misc_feature 609..665
/label=MCS
/note="multiple cloning site"
intron 679..1154
/label=-beta-globin intron
/note="internally truncated intron from human beta-globin"
CDS 1212..1628
/label=gag (truncated)
/note="truncated Moloney murine leukemia virus (MMLV) gag
gene lacking the start codon"
primer_bind 1558..1580
/label=pLXSN 5'
/note="Murine stem cell virus, forward primer. Also called
MSCV"
primer_bind 1596..1612
/label=pBABE 5'
/note="Psi packaging signal, forward primer for pBABE
vectors"
regulatory 2805..2814
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 2811..3485
/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(2941..2959)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind 3209..3228
/label=mCherry-F
/note="mCherry, forward primer"
primer_bind 3403..3426
/label=DsRed1-C
/note="DsRed1, forward primer"
polyA_signal 3620..3741
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3657..3676)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3711..3730
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(3748..4203)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3885..3906)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 4097..4116
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 4230..4334
/gene="bla"
/label=AmpR promoter
promoter 4336..4693
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(4359..4379)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 4544..4679
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 4606..4625
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 4728..5522
/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(4782..4801)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 5392..5411
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(5710..5729)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 5754..5801
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 6130..6718
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6619..6638
/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 attctgcagt cgacggtacc gcgggcccgg
661 gatcctgaga acttcagggt gagtctatgg gacccttgat gttttctttc cccttctttt
721 ctatggttaa gttcatgtca taggaagggg agaagtaaca gggtacacat attgaccaaa
781 tcagggtaat tttgcatttg taattttaaa aaatgctttc ttcttttaat atactttttt
841 gtttatctta tttctaatac tttccctaat ctctttcttt cagggcaata atgatacaat
901 gtatcatgcc tctttgcacc attctaaaga ataacagtga taatttctgg gttaaggcaa
961 tagcaatatt tctgcatata aatatttctg catataaatt gtaactgatg taagaggttt
1021 catattgcta atagcagcta caatccagct accattctgc ttttatttta tggttgggat
1081 aaggctggat tattctgagt ccaagctagg cccttttgct aatcatgttc atacctctta
1141 tcttcctccc acagctcctg ggcaacgtgc tggtctgtgt gctggcccat cactttggca
1201 aagaattcat gggccagact gttaccactc ccttaagttt gaccttaggt cactggaaag
1261 atgtcgagcg gatcgctcac aaccagtcgg tagatgtcaa gaagagacgt tgggttacct
1321 tctgctctgc agaatggcca acctttaacg tcggatggcc gcgagacggc acctttaacc
1381 gagacctcat cacccaggtt aagatcaagg tcttttcacc tggcccgcat ggacacccag
1441 accaggtccc ctacatcgtg acctgggaag ccttggcttt tgacccccct ccctgggtca
1501 agccctttgt acaccctaag cctccgcctc ctcttcctcc atccgccccg tctctccccc
1561 ttgaacctcc tcgttcgacc ccgcctcgat cctcccttta tccagccctc actccttctc
1621 taggcgccaa acctaaacct caagttcttt ctgacagtgg ggggccgctc atcgacctac
1681 ttacagaaga ccccccgcct tatagggacc caagaccacc cccttccgac agggacggaa
1741 atggtggaga agcgacccct gcgggagagg caccggaccc ctccccaatg gcatctcgcc
1801 tacgtgggag acgggagccc cctgtggccg actccactac ctcgcaggca ttccccctcc
1861 gcgcaggagg aaacggacag cttcaatact ggccgttctc ctcttctgac ctttacaact
1921 ggaaaaataa taacccttct ttttctgaag atccaggtaa actgacagct ctgatcgagt
1981 ctgttctcat cacccatcag cccacctggg acgactgtca gcagctgttg gggactctgc
2041 tgaccggaga agaaaaacaa cgggtgctct tagaggctag aaaggcggtg cggggcgatg
2101 atgggcgccc cactcaactg cccaatgaag tcgatgccgc ttttcccctc gagcgcccag
2161 actgggatta caccacccag gcaggtagga accacctagt ccactatcgc cagttgctcc
2221 tagcgggtct ccaaaacgcg ggcagaagcc ccaccaattt ggccaaggta aaaggaataa
2281 cacaagggcc caatgagtct ccctcggcct tcctagagag acttaaggaa gcctatcgca
2341 ggtacactcc ttatgaccct gaggacccag ggcaagaaac taatgtgtct atgtctttca
2401 tttggcagtc tgccccagac attgggagaa agttagagag gttagaagat ttaaaaaaca
2461 agacgcttgg agatttggtt agagaggcag aaaagatctt taataaacga gaaaccccgg
2521 aagaaagaga ggaacgtatc aggagagaaa cagaggaaaa agaagaacgc cgtaggacag
2581 aggatgagca gaaagagaaa gaaagagatc gtaggagaca tagagagatg agcaagctat
2641 tggccactgt cgttagtgga cagaaacagg atagacaggg aggagaacga aggaggtccc
2701 aactcgatcg cgaccagtgt gcctactgca aagaaaaggg gcactgggct aaagattgtc
2761 ccaagaaacc acgaggacct cggggaccaa gaccgcaacc ggtcgccacc atggcctcct
2821 ccgaggacgt catcaaggag ttcatgcgct tcaaggtgcg catggagggc tccgtgaacg
2881 gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg
2941 ccaagctgaa ggtgaccaag ggcggccccc tgcccttcgc ctgggacatc ctgtcccctc
3001 agttccagta cggctccaag gcctacgtga agcaccccgc cgacatcccc gactacttga
3061 agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg
3121 tggtgaccgt gacccaggac tcctccctgc aggacggcga gttcatctac aaggtgaagc
3181 tgcgcggcac caacttcccc tccgacggcc ccgtaatgca gaagaagacc atgggctggg
3241 aggcctccac cgagcggatg taccccgagg acggcgccct gaagggcgag atcaagatga
3301 ggctgaagct gaaggacggc ggccactacg acgccgaggt caagaccacc tacatggcca
3361 agaagcccgt gcagctgccc ggcgcctaca agaccgacat caagctggac atcacctccc
3421 acaacgagga ctacaccatc gtggaacagt acgagcgcgc cgagggccgc cactccaccg
3481 gcgccttgta caagtaaagc ggccgcgact ctagatcata atcagccata ccacatttgt
3541 agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat
3601 gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa
3661 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
3721 caaactcatc aatgtatctt aaggcgtaaa ttgtaagcgt taatattttg ttaaaattcg
3781 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
3841 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
3901 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
3961 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
4021 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
4081 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
4141 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
4201 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
4261 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
4321 gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg
4381 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
4441 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
4501 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
4561 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
4621 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
4681 aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg attgaacaag
4741 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
4801 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
4861 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag
4921 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
4981 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
5041 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
5101 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
5161 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
5221 tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg
5281 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
5341 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
5401 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
5461 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
5521 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
5581 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
5641 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accctagggg
5701 gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat
5761 aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc
5821 ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt
5881 ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa
5941 cgtcggggcg gcaggccctg ccatagcctc aggttactca tatatacttt agattgattt
6001 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
6061 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
6121 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
6181 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
6241 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
6301 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
6361 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
6421 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
6481 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
6541 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
6601 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
6661 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
6721 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
6781 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc at
//
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