| 质粒名称: | pEGFP-LC3 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 21073 |
| 存储实验室: | Tamotsu Yoshimori |
|
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: | 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: | -- |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | microtubule-associated protein 1 light chain 3 beta |
| 物种来源: | R. norvegicus (rat) |
| 插入大小: | 738 bp |
| 基因突变: | First ATG is not included. |
| GenBank编号 | U05784 |
| 启动子 | CMV |
| 标签/融合蛋白: | EGFP (N terminal on backbone) |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | BglII(未破坏) |
| 3' 克隆位点: | EcoRI(未破坏) |
| 5' 测序引物 | EGFP-C |
| 3' 测序引物 | SV40pA-R (5'-GAAATTTGTGATGCTATTGC-3') |
|
文献引用方法 |
|
| 材料和方法部分: | pEGFP-LC3 was a gift from Tamotsu Yoshimori (Addgene plasmid # 21073 ; http://n2t.net/addgene:21073 ; RRID:Addgene_21073) |
| 参考文献部分: |
LC3, a mammalian homologue of yeast Apg8p, is localized in autophagosome membranes after processing. Kabeya Y, Mizushima N, Ueno T, Yamamoto A, Kirisako T, Noda T, Kominami E, Ohsumi Y, Yoshimori T. EMBO J. 2000 Nov 1. 19(21):5720-8. 10.1093/emboj/19.21.5720 PubMed 11060023 |
买家导航
pEGFPLC3载体基本信息
pEGFPLC3载体图谱质粒图谱和多克隆位点信息
pEGFPLC3载体简介
pEGFPLC3载体序列
LOCUS pEGFP-LC3 5452 bp ds-DNA circular SYN 30-AUG-2018
DEFINITION Mammalian expression of rat LC3 fused to EGFP.
ACCESSION .
VERSION .
KEYWORDS pEGFP-LC3
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5452)
AUTHORS Kabeya Y, Mizushima N, Ueno T, Yamamoto A, Kirisako T, Noda T,
Kominami E, Ohsumi Y, Yoshimori T
TITLE LC3, a mammalian homologue of yeast Apg8p, is localized in
autophagosome membranes after processing.
JOURNAL EMBO J. 2000 Nov 1. 19(21):5720-8.
PUBMED 11060023
REFERENCE 2 (bases 1 to 5452)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 30, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5452
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 121..424
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 425..628
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 579..599
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
regulatory 667..676
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 673..1389
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(718..739)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(979..998)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1326..1347
/label=EGFP-C
/note="EGFP, forward primer"
CDS 1408..1830
/codon_start=1
/gene="Map1lc3b"
/product="rat LC3 protein"
/label=LC3
/note="Atg8 family member found on autophagosomes"
/translation="PSEKTFKQRRSFEQRVEDVRLIREQHPTKIPVIIERYKGEKQLPV
LDKTKFLVPDHVNMSELIKIIRRRLQLNANQAFFLLVNGHSMVSVSTPISEVYESERDE
DGFLYMVYASQETFGTALAVTYMSALKATATGREPCL"
polyA_signal 2300..2421
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2337..2356)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 2391..2410
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(2428..2883)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2565..2586)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2777..2796
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 2910..3014
/gene="bla"
/label=AmpR promoter
promoter 3016..3373
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(3039..3059)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 3224..3359
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3286..3305
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3408..4202
/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(3462..3481)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 4072..4091
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(4390..4409)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 4434..4481
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 4810..5398
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5299..5318
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat
61 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
121 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
181 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
241 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
301 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
361 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
421 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
481 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
541 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
601 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
661 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
721 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
781 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
841 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
901 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
961 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
1021 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
1081 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
1141 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
1201 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
1261 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
1321 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
1381 ctgtacaagt ccggactcag atctagaccg tccgagaaga ccttcaaaca gcgccggagc
1441 ttcgaacaaa gagtggaaga tgtccggctc atccgggagc agcaccccac caagatccca
1501 gtgattatag agcgatacaa gggtgagaag cagctgcccg tcctggacaa gaccaagttc
1561 cttgtacctg atcacgtgaa tatgagcgaa ctcatcaaga taattagaag gcgcctgcag
1621 ctcaatgcta accaagcctt cttcctcctg gtgaatgggc acagcatggt gagtgtgtcc
1681 acacccatct ctgaagtgta cgagagcgag agagatgaag acggcttcct gtacatggtc
1741 tatgcctccc aggagacgtt cgggacagca ctggctgtta catacatgtc agctctgaag
1801 gcaacagcaa caggaagaga gccatgcttg tgacagacat acagccactt ccaactaaag
1861 caagcctctg ctccctgcta cctgcatgga gcccactgtg acactcagac catccccggt
1921 cactcactca ctcgtgtctg agaatctcag tgagagctgc ctctgtcacg gacggaagcc
1981 aacacagcca cctctcgacc tggctcccca cagcacccac cctccctgca tgcagctgtc
2041 cctgctaacc cccaatgtta tgttacactg tgtaagtccc cactgctgcc gtgtgtgggt
2101 tgtgtacgtc gtcatgtccc tggtttataa ctatggtgcg aattctgcag tcgacggtac
2161 cgcgggcccg ggatccaccg gatctagata actgatcata atcagccata ccacatttgt
2221 agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat
2281 gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa
2341 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
2401 caaactcatc aatgtatctt aacgcgtaaa ttgtaagcgt taatattttg ttaaaattcg
2461 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
2521 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
2581 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
2641 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
2701 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
2761 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
2821 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
2881 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
2941 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
3001 gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg
3061 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
3121 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
3181 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
3241 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
3301 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
3361 aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg attgaacaag
3421 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
3481 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
3541 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag
3601 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
3661 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
3721 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
3781 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
3841 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
3901 tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg
3961 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
4021 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
4081 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
4141 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
4201 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
4261 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
4321 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accctagggg
4381 gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat
4441 aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc
4501 ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt
4561 ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa
4621 cgtcggggcg gcaggccctg ccatagcctc aggttactca tatatacttt agattgattt
4681 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
4741 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
4801 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
4861 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
4921 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
4981 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
5041 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
5101 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
5161 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
5221 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
5281 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
5341 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
5401 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ct
//
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