| 质粒名称: | mEGFP-PI35P2 |
|---|---|
| 出品公司: | addgene |
| 目录编号: | 92419 |
| 存储实验室: | Geert van den Bogaart |
|
载体骨架基本信息 |
|
| 载体名称: | pEGFP-C1 |
| 载体出品公司: | CloneTech |
| 完整质粒大小: | 5124 bp |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 新霉素（G418） |
| 载体抗性: | Kanamycin |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | MCOLN1/Mouse MCOLN1 residues 1–68; Mucolipin 1, phosphoinositide binding domain. |
| 物种来源: | M. musculus (mouse) |
| 基因突变: | Synthetic gene, codon optimized for human. |
| GenBank编号 | NM_053177.1 NM_053177.1 |
| 启动子: | CMV |
| 标签/融合蛋白: | eGFP (N terminal on backbone) |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | XhoI(未破坏) |
| 3’克隆位点: | BamHI(未破坏) |
| 5’测序引物 | GCCGCCGGGATCAC |
| 3’测序引物 | CCTCTACAAATGTGGTATGGC |
|
文献引用方法 |
|
| 材料和方法部分: | mEGFP-PI35P2 was a gift from Geert van den Bogaart (Addgene plasmid # 92419 ; http://n2t.net/addgene:92419 ; RRID:Addgene_92419) |
| 参考文献部分: | SWAP70 is a universal GEF-like adapter for tethering actin to phagosomes. Baranov MV, Revelo NH, Verboogen DRJ, Ter Beest M, van den Bogaart G. Small GTPases. 2017 May 10:0. doi: 10.1080/21541248.2017.1328302. 10.1080/21541248.2017.1328302 PubMed 2 |
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 mEGFP-PI35P2 5124 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Fluorescent reporter for phosphatidylinositol (3,5)-bisphosphate
(PI(3,5)P2). Mouse MCOLN1 residues 1–68 fused to eGFP..
ACCESSION .
VERSION .
KEYWORDS mEGFP-PI35P2
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5124)
AUTHORS Baranov MV, Revelo NH, Verboogen DRJ, Ter Beest M, van den Bogaart G
TITLE SWAP70 is a universal GEF-like adapter for tethering actin to
phagosomes.
JOURNAL Small GTPases. 2017 May 10:0. doi: 10.1080/21541248.2017.1328302.
PUBMED 28489960
REFERENCE 2 (bases 1 to 5124)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..5124
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 61..364
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 365..568
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 519..539
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
regulatory 607..616
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 613..1329
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
CDS 613..1329
/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(658..679)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(919..938)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1266..1287
/label=EGFP-C
/note="EGFP, forward primer"
polyA_signal 1912..2033
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1949..1968)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 2003..2022
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(2040..2495)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2177..2198)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2389..2408
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 2522..2626
/gene="bla"
/label=AmpR promoter
promoter 2628..2985
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(2651..2671)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 2836..2971
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2898..2917
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3020..3814
/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(3074..3093)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3684..3703
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(4002..4021)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 4046..4093
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 4422..5010
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4911..4930
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
601 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
1321 ctgtacaagt ccggactcag atctcgagct agcgacctga ctatggcaac tccggcgggg
1381 cggcgtgctt ctgaaactga gcgtctgctg actccgaacc ctggatatgg aacccaggtg
1441 ggcacatctc ctgcaccaac cacacccacc gaagaggaag atctgcgccg tcggctgaag
1501 tatttcttta tgagcccatg cgacaaattt cgggcaaaag ggcgtaagcc ttgtaaactg
1561 atgctgcgca ttctgaccat ggcaactccg gccggacgcc gtgcaagcga gaccgagcgt
1621 ctgctgaccc caaaccctgg ctatggtacc caggttggta catcccctgc tccaactacc
1681 ccgaccgagg aagaggatct gcggcgccgt ctgaagtact tctttatgag tccgtgtgac
1741 aagtttcggg caaagggtcg caagccgtgt aagctgatgc tgggatccac cggatctaga
1801 taactgatca taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc
1861 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt
1921 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
1981 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaacgcgta
2041 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
2101 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
2161 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
2221 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
2281 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
2341 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
2401 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
2461 acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt
2521 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
2581 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag
2641 aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
2701 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
2761 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
2821 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
2881 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
2941 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac
3001 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
3061 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
3121 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
3181 cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg
3241 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
3301 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
3361 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
3421 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
3481 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
3541 caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
3601 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
3661 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
3721 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
3781 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
3841 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
3901 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
3961 ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca
4021 ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt
4081 gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca
4141 ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca
4201 agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc
4261 tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
4321 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
4381 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
4441 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
4501 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
4561 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
4621 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
4681 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
4741 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
4801 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
4861 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
4921 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
4981 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
5041 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
5101 gataaccgta ttaccgccat gcat
//
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