| 质粒名称: | pEGFP-2xFYVE |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 140047 |
| 存储实验室: | Harald Stenmark |
|
载体骨架基本信息 |
|
| 载体名称: | pEGFP-C3 |
| 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: | 4219 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: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | tandem HRS FYVE domain,HGS |
| 物种来源: | M. musculus (mouse) |
| 插入大小: | 486 bp |
| 基因突变: | two FYVE domains (amino acids 147-223), linked by QGQGS linker |
| GenBank编号 | 15239 |
| 启动子 | CMV |
| 标签/融合蛋白: | EGFP（插入件上的N端） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | EcoRI in backbone/MfeI in insert（在克隆过程中被破坏） |
| 3' 克隆位点: | SalI（如果已销毁，则未知） |
| 5' 测序引物 | CATGGTCCTGCTGGAGTTCGTG |
| 3' 测序引物 | GTGGTTTGTCCAAACTCATC |
|
文献引用方法 |
|
| 材料和方法部分: | pEGFP-2xFYVE was a gift from Harald Stenmark (Addgene plasmid # 140047 ; http://n2t.net/addgene:140047 ; RRID:Addgene_140047) |
| 参考文献部分: | Localization of phosphatidylinositol 3-phosphate in yeast and mammalian cells. Gillooly DJ, Morrow IC, Lindsay M, Gould R, Bryant NJ, Gaullier JM, Parton RG, Stenmark H. EMBO J. 2000 Sep 1;19(17):4577-88. doi: 10.1093/emboj/19.17.4577. 10.1093/emboj/19.17.4577 PubMed 10970851 |
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pEGFP-2xFYVE载体 pEGFP2xFYVE质粒基本信息
pEGFP-2xFYVE质粒图谱 pEGFP2xFYVE载体图谱和pEGFP-2xFYVE载体序列 pEGFP2xFYVE质粒序列多克隆位点信息
pEGFP-2xFYVE质粒 pEGFP2xFYVE载体简介
pEGFP-2xFYVE质粒序列 pEGFP2xFYVE载体序列
LOCUS pEGFP-2xFYVE 5208 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Expresses GFP-tagged HRS 2xFYVE in mammalian cells.
ACCESSION .
VERSION .
KEYWORDS pEGFP-2xFYVE
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5208)
AUTHORS Gillooly DJ, Morrow IC, Lindsay M, Gould R, Bryant NJ, Gaullier JM,
Parton RG, Stenmark H
TITLE Localization of phosphatidylinositol 3-phosphate in yeast and
mammalian cells.
JOURNAL EMBO J. 2000 Sep 1;19(17):4577-88. doi: 10.1093/emboj/19.17.4577.
PUBMED 10970851
REFERENCE 2 (bases 1 to 5208)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5208
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 60..363
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 364..567
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 518..538
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
regulatory 606..615
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 612..1328
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
CDS 612..1328
/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(657..678)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(918..937)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1265..1286
/label=EGFP-C
/note="EGFP, forward primer"
polyA_signal 1995..2116
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2032..2051)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 2086..2105
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(2123..2578)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2260..2281)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2472..2491
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 2605..2709
/gene="bla"
/label=AmpR promoter
promoter 2711..3068
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(2734..2754)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 2919..3054
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2981..3000
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3103..3897
/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(3157..3176)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3767..3786
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(4085..4104)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 4129..4176
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 4505..5093
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4994..5013
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
61 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
121 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
181 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
241 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
301 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
361 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga
421 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
481 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta
541 cggtgggagg tctatataag cagagctggt ttagtgaacc gtcagatccg ctagcgctac
601 cggtcgccac catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
661 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg
721 atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc
781 cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg
841 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
901 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
961 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
1021 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca
1081 agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg
1141 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
1201 ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg
1261 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
1321 tgtacaagta ctcagatctc gagctcaagc ttcgaattga attcgaaagt gatgccatgt
1381 tcgctgctga aagagcccct gactgggtgg atgctgagga atgccatcgg tgcagagtac
1441 agtttggggt ggtgacccgc aagcatcact gccgagcatg tgggcagatc ttctgtggca
1501 agtgctcctc caagtactcc accatcccca agttcggcat tgagaaggag gtgcgcgtgt
1561 gtgagccctg ctatgagcag ctgaacaaga aggcacaagg acaaggatcc gaaagtgatg
1621 ccatgttcgc tgctgaaaga gcccctgact gggtggatgc tgaggaatgc catcggtgca
1681 gagtacagtt tggggtggtg acccgcaagc atcactgccg agcatgtggg cagatcttct
1741 gtggcaagtg ctcctccaag tactccacca tccccaagtt cggcattgag aaggaggtgc
1801 gcgtgtgtga gccctgctat gagcagctga acaagaaggc ataagtcgac ggtaccgcgg
1861 gcccgggatc caccggatct agataactga tcataatcag ccataccaca tttgtagagg
1921 ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg
1981 caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
2041 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
2101 tcatcaatgt atcttaacgc gtaaattgta agcgttaata ttttgttaaa attcgcgtta
2161 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
2221 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
2281 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
2341 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
2401 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
2461 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
2521 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca
2581 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
2641 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
2701 aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa
2761 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
2821 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
2881 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
2941 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
3001 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
3061 tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
3121 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
3181 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
3241 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg
3301 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
3361 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
3421 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
3481 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
3541 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
3601 ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg
3661 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
3721 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
3781 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
3841 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
3901 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
3961 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
4021 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc
4081 taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa
4141 gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg
4201 gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt
4261 ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg
4321 gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac
4381 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
4441 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
4501 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
4561 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
4621 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
4681 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
4741 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
4801 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
4861 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
4921 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
4981 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
5041 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
5101 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
5161 ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcatt
//
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