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 |
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 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 //