| 质粒名称: | EGFP-LC3 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 11546 |
| 存储实验室: | Karla Kirkegaard |
|
载体骨架基本信息 |
|
| 载体名称: | pEGFP-C3 |
| 载体出品公司: | Clontech |
| 空载体大小: | 4727 bp |
| 完整质粒大小: | 5113bp |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 新霉素（G418） |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 未知 |
|
插入基因信息 |
|
| 插入基因名称: | LC3B / NM_022818 / Human homolog of yeast ATG8p |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 380bp |
| GenBank编号 | NM_022818 |
| 启动子: | CMV |
| 标签/融合蛋白: | EGFP (N terminal on insert) |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | EcoRI(未破坏) |
| 3’克隆位点: | EcoRI (未破坏) |
| 5’测序引物 | CMV-F |
| 3’测序引物 | EBV Reverse |
|
文献引用方法 |
|
| 材料和方法部分: |
EGFP-LC3 was a gift from Karla Kirkegaard (Addgene plasmid # 11546 ; http://n2t.net/addgene:11546 ; RRID:Addgene_11546)
|
| 参考文献部分: | Subversion of cellular autophagosomal machinery by RNA viruses. Jackson WT, Giddings TH, Taylor MP, Mulinyawe S, Rabinovitch M, Kopito RR, Kirkegaard K. PLoS Biol. 2005 May . 3(5):e156. 10.1371/journal.pbio.0030156 PubMed 15884975 |
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 EGFP-LC3 5113 bp ds-DNA circular SYN 16-AUG-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS EGFP-LC3
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5113)
AUTHORS Jackson WT, Giddings TH, Taylor MP, Mulinyawe S, Rabinovitch M,
Kopito RR, Kirkegaard K
TITLE Subversion of cellular autophagosomal machinery by RNA viruses.
JOURNAL PLoS Biol. 2005 May . 3(5):e156.
PUBMED 15884975
REFERENCE 2 (bases 1 to 5113)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 16, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..5113
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter complement(24..381)
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 38..173
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind complement(92..111)
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
primer_bind 338..358
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter complement(383..487)
/gene="bla"
/label=AmpR promoter
rep_origin 514..969
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(601..620)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 811..832
/label=F1ori-F
/note="F1 origin, forward primer"
polyA_signal 976..1097
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(987..1006)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 1041..1060
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
CDS complement(1261..1635)
/codon_start=1
/gene="MAP1LC3B"
/product="human LC3 protein"
/label=LC3B
/note="Atg8 family member found on autophagosomes"
/translation="MPSEKTFKQRRTFEQRVEDVRLIREQHPTKIPVIIERYKGEKQLP
VLDKTKFLVPDHVNMSELIKIIRRRLQLNANQAFFLLVNGHSMVSVSTPISEVYESEKD
EDGFLYMVYASQETFGMKLSV"
CDS complement(1669..2385)
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
CDS complement(1669..2385)
/codon_start=1
/product="the original enhanced GFP (Yang et al., 1996)"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(1711..1732)
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind 2060..2079
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 2319..2340
/label=EGFP-N
/note="EGFP, reverse primer"
regulatory 2382..2391
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
promoter complement(2430..2633)
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind complement(2459..2479)
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
enhancer 2634..2937
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
rep_origin complement(3112..3700)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3192..3211)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
polyA_signal 4029..4076
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
primer_bind 4101..4120
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
CDS complement(4308..5102)
/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(4419..4438)
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind 5029..5048
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
ORIGIN
1 ctcatcctgt ctcttgatcg atctttgcaa aagcctaggc ctccaaaaaa gcctcctcac
61 tacttctgga atagctcaga ggccgaggcg gcctcggcct ctgcataaat aaaaaaaatt
121 agtcagccat ggggcggaga atgggcggaa ctgggcggag ttaggggcgg gatgggcgga
181 gttaggggcg ggactatggt tgctgactaa ttgagatgca tgctttgcat acttctgcct
241 gctggggagc ctggggactt tccacacctg gttgctgact aattgagatg catgctttgc
301 atacttctgc ctgctgggga gcctggggac tttccacacc ctaactgaca cacattccac
361 agctggttct ttccgcctca ggactcttcc tttttcaata ttattgaagc atttatcagg
421 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
481 ttccgcgcac atttccccga aaagtgccac ctgacgcgcc ctgtagcggc gcattaagcg
541 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
601 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
661 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
721 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
781 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
841 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
901 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc
961 ttacaattta cgcgttaaga tacattgatg agtttggaca aaccacaact agaatgcagt
1021 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
1081 gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg
1141 aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtatg gctgattatg
1201 atcagttatc tagatccggt ggatcccggg cccgcggtac cgtcgactgc agaattctta
1261 cactgacaat ttcatcccga acgtctcctg ggaggcatag accatgtaca ggaatccatc
1321 ttcatctttc tcactctcat acacctctga gattggtgtg gagacgctga ccatgctgtg
1381 tccgttcacc aacaggaaga aggcctgatt agcattgagc tgtaagcgcc ttctaattat
1441 cttgatgagc tcactcatgt tgacatggtc aggtacaagg aactttgttt tatccagaac
1501 aggaagctgc ttctcaccct tgtatcgttc tattatcacc gggattttgg ttggatgctg
1561 ctctcgaata agtcggacat cttctactct ttgttcgaag gtgcggcgct gcttgaaggt
1621 cttctccgac ggcatgggaa ttcgaagctt gagctcgaga tctgagtact tgtacagctc
1681 gtccatgccg agagtgatcc cggcggcggt cacgaactcc agcaggacca tgtgatcgcg
1741 cttctcgttg gggtctttgc tcagggcgga ctgggtgctc aggtagtggt tgtcgggcag
1801 cagcacgggg ccgtcgccga tgggggtgtt ctgctggtag tggtcggcga gctgcacgct
1861 gccgtcctcg atgttgtggc ggatcttgaa gttcaccttg atgccgttct tctgcttgtc
1921 ggccatgata tagacgttgt ggctgttgta gttgtactcc agcttgtgcc ccaggatgtt
1981 gccgtcctcc ttgaagtcga tgcccttcag ctcgatgcgg ttcaccaggg tgtcgccctc
2041 gaacttcacc tcggcgcggg tcttgtagtt gccgtcgtcc ttgaagaaga tggtgcgctc
2101 ctggacgtag ccttcgggca tggcggactt gaagaagtcg tgctgcttca tgtggtcggg
2161 gtagcggctg aagcactgca cgccgtaggt cagggtggtc acgagggtgg gccagggcac
2221 gggcagcttg ccggtggtgc agatgaactt cagggtcagc ttgccgtagg tggcatcgcc
2281 ctcgccctcg ccggacacgc tgaacttgtg gccgtttacg tcgccgtcca gctcgaccag
2341 gatgggcacc accccggtga acagctcctc gcccttgctc accatggtgg cgaccggtag
2401 cgctagcgga tctgacggtt cactaaacca gctctgctta tatagacctc ccaccgtaca
2461 cgcctaccgc ccatttgcgt caatggggcg gagttgttac gacattttgg aaagtcccgt
2521 tgattttggt gccaaaacaa actcccattg acgtcaatgg ggtggagact tggaaatccc
2581 cgtgagtcaa accgctatcc acgcccattg atgtactgcc aaaaccgcat caccatggta
2641 atagcgatga ctaatacgta gatgtactgc caagtaggaa agtcccataa ggtcatgtac
2701 tgggcataat gccaggcggg ccatttaccg tcattgacgt caataggggg cgtacttggc
2761 atatgataca cttgatgtac tgccaagtgg gcagtttacc gtaaatactc cacccattga
2821 cgtcaatgga aagtccctat tggcgttact atgggaacat acgtcattat tgacgtcaat
2881 gggcgggggt cgttgggcgg tcagccaggc gggccattta ccgtaagtta tgtaacgcgg
2941 aactccatat atgggctatg aactaatgac cccgtaattg attactatta ataactaatg
3001 catggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
3061 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
3121 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
3181 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
3241 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
3301 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
3361 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
3421 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
3481 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
3541 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
3601 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
3661 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
3721 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
3781 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
3841 aagtatatat gagtaacctg aggctatggc agggcctgcc gccccgacgt tggctgcgag
3901 ccctgggcct tcacccgaac ttggggggtg gggtggggaa aaggaagaaa cgcgggcgta
3961 ttggccccaa tggggtctcg gtggggtatc gacagagtgc cagccctggg accgaacccc
4021 gcgtttatga acaaacgacc caacaccgtg cgttttattc tgtcttttta ttgccgtcat
4081 agcgcgggtt ccttccggta ttgtctcctt ccgtgtttca gttagcctcc ccctagggtg
4141 ggcgaagaac tccagcatga gatccccgcg ctggaggatc atccagccgg cgtcccggaa
4201 aacgattccg aagcccaacc tttcatagaa ggcggcggtg gaatcgaaat ctcgtgatgg
4261 caggttgggc gtcgcttggt cggtcatttc gaaccccaga gtcccgctca gaagaactcg
4321 tcaagaaggc gatagaaggc gatgcgctgc gaatcgggag cggcgatacc gtaaagcacg
4381 aggaagcggt cagcccattc gccgccaagc tcttcagcaa tatcacgggt agccaacgct
4441 atgtcctgat agcggtccgc cacacccagc cggccacagt cgatgaatcc agaaaagcgg
4501 ccattttcca ccatgatatt cggcaagcag gcatcgccat gggtcacgac gagatcctcg
4561 ccgtcgggca tgctcgcctt gagcctggcg aacagttcgg ctggcgcgag cccctgatgc
4621 tcttcgtcca gatcatcctg atcgacaaga ccggcttcca tccgagtacg tgctcgctcg
4681 atgcgatgtt tcgcttggtg gtcgaatggg caggtagccg gatcaagcgt atgcagccgc
4741 cgcattgcat cagccatgat ggatactttc tcggcaggag caaggtgaga tgacaggaga
4801 tcctgccccg gcacttcgcc caatagcagc cagtcccttc ccgcttcagt gacaacgtcg
4861 agcacagctg cgcaaggaac gcccgtcgtg gccagccacg atagccgcgc tgcctcgtct
4921 tgcagttcat tcagggcacc ggacaggtcg gtcttgacaa aaagaaccgg gcgcccctgc
4981 gctgacagcc ggaacacggc ggcatcagag cagccgattg tctgttgtgc ccagtcatag
5041 ccgaatagcc tctccaccca agcggccgga gaacctgcgt gcaatccatc ttgttcaatc
5101 atgcgaaacg atc
//
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