| 质粒名称: | ptfLC3 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 21074 |
| 存储实验室: | Tamotsu Yoshimori |
|
载体骨架基本信息 |
|
| 载体名称: | pEGFP-C1+mRFP |
| 载体抗性: | 卡那霉素 |
| 空载体大小: | 5200 bp |
| 完整质粒大小: | 5938 bp |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 新霉素（G418） |
| 高拷贝/低拷贝: | 未知 |
| 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: | Map1lc3b，LC3，LC3b |
| GenBank编号 | U05784 |
| 标签/融合蛋白: |
mRFP (N terminal on backbone)
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 | N/A |
|
文献引用方法 |
|
| 材料和方法部分: | ptfLC3 was a gift from Tamotsu Yoshimori (爱迪基因 plasmid # 21074) |
| 参考文献部分: | Dissection of the autophagosome maturation process by a novel reporter protein, tandem fluorescent-tagged LC3. Kimura S, Noda T, Yoshimori T. Autophagy. . 3(5):452-60. PubMed 17534139 |
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 ptfLC3 6136 bp ds-DNA circular SYN 10-JAN-2018
DEFINITION Mammalian expression of rat LC3 fused to mRFP and EGFP.
ACCESSION .
VERSION .
KEYWORDS ptfLC3
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6136)
AUTHORS Kimura S, Noda T, Yoshimori T
TITLE Dissection of the autophagosome maturation process by a novel
reporter protein, tandem fluorescent-tagged LC3.
JOURNAL Autophagy. . 3(5):452-60.
PUBMED 17534139
COMMENT This file is created by Vector NTI
http://www.biofeng.com/
COMMENT ORIGDB|GenBank
COMMENT VNTAUTHORNAME|biofeng.com|
COMMENT VNTNAME|ptfLC3|
FEATURES Location/Qualifiers
source 1..6136
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 200..220
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter complement(245..349)
/gene="bla"
/label=AmpR promoter
rep_origin 376..831
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(463..482)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 673..694
/label=F1ori-F
/note="F1 origin, forward primer"
polyA_signal 838..959
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(849..868)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 903..922
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
CDS complement(1429..1851)
/codon_start=1
/gene="Map1lc3b"
/product="rat LC3 protein"
/label=LC3
/note="Atg8 family member found on autophagosomes"
/translation="PSEKTFKQRRSFEQRVEDVRLIREQHPTKIPVIIERYKGEKQLPV
LDKTKFLVPDHVNMSELIKIIRRRLQLNANQAFFLLVNGHSMVSVSTPISEVYESERDE
DGFLYMVYASQETFGTALAVTYMSALKATATGREPCL"
CDS complement(1870..2586)
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(1912..1933)
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind 2261..2280
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 2520..2541
/label=EGFP-N
/note="EGFP, reverse primer"
regulatory 2583..2592
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS complement(2605..3279)
/codon_start=1
/product="monomeric derivative of DsRed (Campbell et al.,
2002)"
/label=mRFP1
/note="mammalian codon-optimized"
/translation="MAFSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
TGA"
primer_bind complement(2664..2687)
/label=DsRed1-C
/note="DsRed1, forward primer"
primer_bind complement(2862..2881)
/label=mCherry-F
/note="mCherry, forward primer"
primer_bind 3131..3149
/label=mCherry-R
/note="mCherry, reverse primer"
regulatory 3276..3285
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
promoter complement(3315..3518)
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind complement(3344..3364)
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
enhancer 3519..3822
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
rep_origin complement(3997..4585)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4077..4096)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
polyA_signal 4914..4961
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
primer_bind 4986..5005
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
CDS complement(5193..5987)
/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(5304..5323)
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind 5914..5933
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
promoter complement(6022..243)
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 6036..35
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind complement(6090..6109)
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
ORIGIN
1 gaatgggcgg aactgggcgg agttaggggc gggatgggcg gagttagggg cgggactatg
61 gttgctgact aattgagatg catgctttgc atacttctgc ctgctgggga gcctggggac
121 tttccacacc tggttgctga ctaattgaga tgcatgcttt gcatacttct gcctgctggg
181 gagcctgggg actttccaca ccctaactga cacacattcc acagctggtt ctttccgcct
241 caggactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
301 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
361 gaaaagtgcc acctgacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
421 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
481 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
541 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
601 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
661 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
721 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
781 tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt tacgcgttaa
841 gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt
901 gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta
961 acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt
1021 aaagcaagta aaacctctac aaatgtggta tggctgatta tgatcagtta tctagatccg
1081 gtggatcccg ggcccgcggt accgtcgact gcagaattcg caccatagtt ataaaccagg
1141 gacatgacga cgtacacaac ccacacacgg cagcagtggg gacttacaca gtgtaacata
1201 acattggggg ttagcaggga cagctgcatg cagggagggt gggtgctgtg gggagccagg
1261 tcgagaggtg gctgtgttgg cttccgtccg tgacagaggc agctctcact gagattctca
1321 gacacgagtg agtgagtgac cggggatggt ctgagtgtca cagtgggctc catgcaggta
1381 gcagggagca gaggcttgct ttagttggaa gtggctgtat gtctgtcaca agcatggctc
1441 tcttcctgtt gctgttgcct tcagagctga catgtatgta acagccagtg ctgtcccgaa
1501 cgtctcctgg gaggcataga ccatgtacag gaagccgtct tcatctctct cgctctcgta
1561 cacttcagag atgggtgtgg acacactcac catgctgtgc ccattcacca ggaggaagaa
1621 ggcttggtta gcattgagct gcaggcgcct tctaattatc ttgatgagtt cgctcatatt
1681 cacgtgatca ggtacaagga acttggtctt gtccaggacg ggcagctgct tctcaccctt
1741 gtatcgctct ataatcactg ggatcttggt ggggtgctgc tcccggatga gccggacatc
1801 ttccactctt tgttcgaagc tccggcgctg tttgaaggtc ttctcggacg gtctagatct
1861 gagtccggac ttgtacagct cgtccatgcc gagagtgatc ccggcggcgg tcacgaactc
1921 cagcaggacc atgtgatcgc gcttctcgtt ggggtctttg ctcagggcgg actgggtgct
1981 caggtagtgg ttgtcgggca gcagcacggg gccgtcgccg atgggggtgt tctgctggta
2041 gtggtcggcg agctgcacgc tgccgtcctc gatgttgtgg cggatcttga agttcacctt
2101 gatgccgttc ttctgcttgt cggccatgat atagacgttg tggctgttgt agttgtactc
2161 cagcttgtgc cccaggatgt tgccgtcctc cttgaagtcg atgcccttca gctcgatgcg
2221 gttcaccagg gtgtcgccct cgaacttcac ctcggcgcgg gtcttgtagt tgccgtcgtc
2281 cttgaagaag atggtgcgct cctggacgta gccttcgggc atggcggact tgaagaagtc
2341 gtgctgcttc atgtggtcgg ggtagcggct gaagcactgc acgccgtagg tcagggtggt
2401 cacgagggtg ggccagggca cgggcagctt gccggtggtg cagatgaact tcagggtcag
2461 cttgccgtag gtggcatcgc cctcgccctc gccggacacg ctgaacttgt ggccgtttac
2521 gtcgccgtcc agctcgacca ggatgggcac caccccggtg aacagctcct cgcccttgct
2581 caccatggtg gcgaccggtc caccggcgcc ggtggagtgg cggccctcgg cgcgctcgta
2641 ctgttccacg atggtgtagt cctcgttgtg ggaggtgatg tccagcttga tgtcggtctt
2701 gtaggcgccg ggcagctgca cgggcttctt ggccatgtag gtggtcttga cctcggcgtc
2761 gtagtggccg ccgtccttca gcttcagcct catcttgatc tcgcccttca gggcgccgtc
2821 ctcggggtac atccgctcgg tggaggcctc ccagcccatg gtcttcttct gcattacggg
2881 gccgtcggag gggaagttgg tgccgcgcag cttcaccttg tagatgaact cgccgtcctg
2941 cagggaggag tcctgggtca cggtcaccac gccgccgtcc tcgaagttca tcacgcgctc
3001 ccacttgaag ccctcgggga aggacagctt caagtagtcg gggatgtcgg cggggtgctt
3061 cacgtaggcc ttggagccgt actggaactg aggggacagg atgtcccagg cgaagggcag
3121 ggggccgccc ttggtcacct tcagcttggc ggtctgggtg ccctcgtagg ggcggccctc
3181 gccctcgccc tcgatctcga actcgtggcc gttcacggag ccctccatgc gcaccttgaa
3241 gcgcatgaac tccttgatga cgtcctcgga gaaggccatg gtggccgcta gcggatctga
3301 cggttcacta aaccagctct gcttatatag acctcccacc gtacacgcct accgcccatt
3361 tgcgtcaatg gggcggagtt gttacgacat tttggaaagt cccgttgatt ttggtgccaa
3421 aacaaactcc cattgacgtc aatggggtgg agacttggaa atccccgtga gtcaaaccgc
3481 tatccacgcc cattgatgta ctgccaaaac cgcatcacca tggtaatagc gatgactaat
3541 acgtagatgt actgccaagt aggaaagtcc cataaggtca tgtactgggc ataatgccag
3601 gcgggccatt taccgtcatt gacgtcaata gggggcgtac ttggcatatg atacacttga
3661 tgtactgcca agtgggcagt ttaccgtaaa tactccaccc attgacgtca atggaaagtc
3721 cctattggcg ttactatggg aacatacgtc attattgacg tcaatgggcg ggggtcgttg
3781 ggcggtcagc caggcgggcc atttaccgta agttatgtaa cgcggaactc catatatggg
3841 ctatgaacta atgaccccgt aattgattac tattaataac taatgcatgg cggtaatacg
3901 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
3961 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
4021 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
4081 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
4141 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
4201 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
4261 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
4321 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
4381 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
4441 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
4501 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
4561 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
4621 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
4681 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
4741 acctgaggct atggcagggc ctgccgcccc gacgttggct gcgagccctg ggccttcacc
4801 cgaacttggg gggtggggtg gggaaaagga agaaacgcgg gcgtattggc cccaatgggg
4861 tctcggtggg gtatcgacag agtgccagcc ctgggaccga accccgcgtt tatgaacaaa
4921 cgacccaaca ccgtgcgttt tattctgtct ttttattgcc gtcatagcgc gggttccttc
4981 cggtattgtc tccttccgtg tttcagttag cctcccccta gggtgggcga agaactccag
5041 catgagatcc ccgcgctgga ggatcatcca gccggcgtcc cggaaaacga ttccgaagcc
5101 caacctttca tagaaggcgg cggtggaatc gaaatctcgt gatggcaggt tgggcgtcgc
5161 ttggtcggtc atttcgaacc ccagagtccc gctcagaaga actcgtcaag aaggcgatag
5221 aaggcgatgc gctgcgaatc gggagcggcg ataccgtaaa gcacgaggaa gcggtcagcc
5281 cattcgccgc caagctcttc agcaatatca cgggtagcca acgctatgtc ctgatagcgg
5341 tccgccacac ccagccggcc acagtcgatg aatccagaaa agcggccatt ttccaccatg
5401 atattcggca agcaggcatc gccatgggtc acgacgagat cctcgccgtc gggcatgctc
5461 gccttgagcc tggcgaacag ttcggctggc gcgagcccct gatgctcttc gtccagatca
5521 tcctgatcga caagaccggc ttccatccga gtacgtgctc gctcgatgcg atgtttcgct
5581 tggtggtcga atgggcaggt agccggatca agcgtatgca gccgccgcat tgcatcagcc
5641 atgatggata ctttctcggc aggagcaagg tgagatgaca ggagatcctg ccccggcact
5701 tcgcccaata gcagccagtc ccttcccgct tcagtgacaa cgtcgagcac agctgcgcaa
5761 ggaacgcccg tcgtggccag ccacgatagc cgcgctgcct cgtcttgcag ttcattcagg
5821 gcaccggaca ggtcggtctt gacaaaaaga accgggcgcc cctgcgctga cagccggaac
5881 acggcggcat cagagcagcc gattgtctgt tgtgcccagt catagccgaa tagcctctcc
5941 acccaagcgg ccggagaacc tgcgtgcaat ccatcttgtt caatcatgcg aaacgatcct
6001 catcctgtct cttgatcgat ctttgcaaaa gcctaggcct ccaaaaaagc ctcctcacta
6061 cttctggaat agctcagagg ccgaggcggc ctcggcctct gcataaataa aaaaaattag
6121 tcagccatgg ggcgga
//
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