| 质粒名称: | pEGFP-LC3 (human) |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 24920 |
| 存储实验室: | Toren Finkel |
|
载体骨架基本信息 |
|
| 载体名称: | pEGFP-C2 |
| 载体抗性: | 卡那霉素 |
| 空载体大小: | 4700 bp |
| 载体类型: | 哺乳细胞表达 |
| 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: | LC3 |
| 物种来源: | H. sapiens (human) |
| 基因又名: | microtubule-associated protein 1 light chain 3 |
| GenBank编号 | BC067797 |
| 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: | BamHI (未破坏) |
| 5' 测序引物 | CMV-F, EGFP-C |
| 3' 测序引物 | SV40pA-R |
|
文献引用方法 |
|
| 材料和方法部分: | pEGFP-LC3 (human) was a gift from Toren Finkel (爱迪基因 plasmid # 24920) |
| 参考文献部分: | A role for the NAD-dependent deacetylase Sirt1 in the regulation of autophagy. Lee IH, Cao L, Mostoslavsky R, Lombard DB, Liu J, Bruns NE, Tsokos M, Alt FW, Finkel T. Proc Natl Acad Sci U S A. 2008 Mar 4. 105(9):3374-9.10.1073/pnas.0712145105 PubMed 18296641 |
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pEGFP-LC3 (human)载体质粒基本信息
pEGFP-LC3 (human)质粒图谱载体图谱和pEGFP-LC3 (human)载体序列质粒序列多克隆位点信息
质粒图谱.png)
pEGFP-LC3 (human)质粒载体简介
pEGFP-LC3 (human)质粒序列载体序列
LOCUS pEGFP-LC3 (human) 5093 bp ds-DNA circular SYN 10-JAN-2018
DEFINITION Mammalian expression of LC3 fused to EGFP.
ACCESSION .
VERSION .
KEYWORDS pEGFP-LC3 (human)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5093)
AUTHORS Lee IH, Cao L, Mostoslavsky R, Lombard DB, Liu J, Bruns NE, Tsokos
M, Alt FW, Finkel T
TITLE A role for the NAD-dependent deacetylase Sirt1 in the regulation of
autophagy.
JOURNAL Proc Natl Acad Sci U S A. 2008 Mar 4. 105(9):3374-9.
PUBMED 18296641
COMMENT This file is created by Vector NTI
http://www.biofeng.com/
COMMENT ORIGDB|GenBank
COMMENT VNTAUTHORNAME|biofeng.com|
COMMENT VNTNAME|pEGFP-LC3 (human)|
FEATURES Location/Qualifiers
source 1..5093
/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..1336
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(665..686)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(926..945)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1273..1294
/label=EGFP-C
/note="EGFP, forward primer"
CDS 1382..1756
/codon_start=1
/gene="MAP1LC3B"
/product="human LC3 protein"
/label=LC3B
/note="Atg8 family member found on autophagosomes"
/translation="MPSEKTFKQRRTFEQRVEDVRLIREQHPTKIPVIIERYKGEKQLP
VLDKTKFLVPDHVNMSELIKIIRRRLQLNANQAFFLLVNGHSMVSVSTPISEVYESEKD
EDGFLYMVYASQETFGMKLSV"
polyA_signal 1888..2009
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1925..1944)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 1979..1998
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(2016..2471)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2153..2174)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2365..2384
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 2498..2602
/gene="bla"
/label=AmpR promoter
promoter 2604..2961
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(2627..2647)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 2812..2947
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2874..2893
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 2996..3790
/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(3050..3069)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3660..3679
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(3978..3997)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 4022..4069
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 4398..4986
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4887..4906
/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 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
661 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
721 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
781 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg
841 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
901 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
961 gttcgagggc gacaccctgg tgaaccgcat cgagctaaag ggcatcgact tcaaggagga
1021 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat
1081 ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga
1141 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
1201 gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga
1261 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
1321 ggacgagctg tacaagtccg gccggactca gatctcgagc tcaagcttcg aattcccgac
1381 catgccgtcg gagaagacct tcaagcagcg ccgcaccttc gaacaaagag tagaagatgt
1441 ccgacttatt cgagagcagc atccaaccaa aatcccggtg ataatagaac gatacaaggg
1501 tgagaagcag cttcctgttc tggataaaac aaagttcctt gtacctgacc atgtcaacat
1561 gagtgagctc atcaagataa ttagaaggcg cttacagctc aatgctaatc aggccttctt
1621 cctgttggtg aacggacaca gcatggtcag cgtctccaca ccaatctcag aggtgtatga
1681 gagtgagaaa gatgaagatg gattcctgta catggtctat gcctcccagg agacgttcgg
1741 gatgaaattg tcagtgcagg atccaccgga tctagataac tgatcataat cagccatacc
1801 acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa
1861 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa
1921 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
1981 ggtttgtcca aactcatcaa tgtatcttaa cgcgtaaatt gtaagcgtta atattttgtt
2041 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
2101 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
2161 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
2221 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
2281 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
2341 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
2401 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
2461 acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
2521 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
2581 ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag
2641 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
2701 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
2761 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
2821 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
2881 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
2941 ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat
3001 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
3061 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
3121 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga
3181 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
3241 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
3301 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
3361 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
3421 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
3481 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga
3541 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
3601 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
3661 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
3721 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
3781 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
3841 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
3901 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
3961 cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg
4021 acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg
4081 gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac
4141 gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc
4201 gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag
4261 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
4321 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
4381 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
4441 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
4501 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
4561 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
4621 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
4681 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
4741 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
4801 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
4861 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
4921 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
4981 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
5041 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgc
//
