| 质粒名称: | GFP-LiMETER-v1 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 113933 |
| 存储实验室: | Yubin Zhou |
|
载体骨架基本信息 |
|
| 载体名称: | pcDNA3.1 |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 5400 bp |
| 完整质粒大小: | 6900 bp |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | -- |
| 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: | LOV2 and Rit polybasic domain (PB),LiMETER-V1 |
| 物种来源: | Synthetic |
| 插入大小: | 1500 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | CMV |
| 标签/融合蛋白: | GFP（插入件上的N端子） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | NheI (未破坏) |
| 3' 克隆位点: | XbaI(未破坏) |
| 5' 测序引物 | CMVf: CGC AAA TGG GCG GTA GGC GTG |
| 3' 测序引物 | BGHrev: TAG AAG GCA CAG TCG AGG |
|
文献引用方法 |
|
| 材料和方法部分: | GFP-LiMETER-v1 was a gift from Yubin Zhou (Addgene plasmid # 113933 ; http://n2t.net/addgene:113933 ; RRID:Addgene_113933) |
| 参考文献部分: | Proteomic mapping of ER-PM junctions identifies STIMATE as a regulator of Ca(2+) influx. Jing J, He L, Sun A, Quintana A, Ding Y, Ma G, Tan P, Liang X, Zheng X, Chen L, Shi X, Zhang SL, Zhong L, Huang Y, Dong MQ, Walker CL, Hogan PG, Wang Y, Zhou Y. Nat Cell Biol. 2015 Oct;17(10):1339-47. doi: 10.1038/ncb3234. Epub 2015 Aug 31. 10.1038/ncb3234 PubMed 26322679 |
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 GFP-LiMETER-v1 6907 bp ds-DNA circular SYN 18-OCT-2018
DEFINITION Express an optogenetic tether to label and interrogate ER-plasma
membrane contact sites (MCS); visualized by GFP.
ACCESSION .
VERSION .
KEYWORDS GFP-LiMETER-v1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6907)
AUTHORS Jing J, He L, Sun A, Quintana A, Ding Y, Ma G, Tan P, Liang X, Zheng
X, Chen L, Shi X, Zhang SL, Zhong L, Huang Y, Dong MQ, Walker CL,
Hogan PG, Wang Y, Zhou Y
TITLE Proteomic mapping of ER-PM junctions identifies STIMATE as a
regulator of Ca(2+) influx.
JOURNAL Nat Cell Biol. 2015 Oct;17(10):1339-47. doi: 10.1038/ncb3234. Epub
2015 Aug 31.
PUBMED 26322679
REFERENCE 2 (bases 1 to 6907)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Oct 18, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6907
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 91..470
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 471..674
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 625..645
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 719..738
/label=T7
/note="T7 promoter, forward primer"
promoter 719..737
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 877..1593
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(922..943)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(1183..1202)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1530..1551
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind complement(2354..2371)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 2360..2584
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 2630..3058
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2717..2736)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2927..2948
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(3067..3087)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 3072..3401
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 3252..3387
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3314..3333
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3468..4262
/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
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(3522..3541)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 4132..4151
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 4436..4557
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4473..4492)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4527..4546
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(4606..4622)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4606..4622)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4619..4641)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4630..4646
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(4654..4684)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4699..4720
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4837..4854)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5008..5596)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5088..5107)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5767..6627)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 6390..6409
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6628..6732)
/gene="bla"
/label=AmpR promoter
primer_bind complement(6807..6826)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
ORIGIN
1 ggcaaggctt gaccgacaat tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc
61 ttcgcgatgt acgggccaga tatacgcgtt gacattgatt attgactagt tattaatagt
121 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
181 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
241 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
301 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
361 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
421 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt
481 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
541 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
601 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
661 atataagcag agctctctgg ctaactagag aacccactgc ttactggctt atcgaaatta
721 atacgactca ctatagggag acccaagctg gctagcaagc ttatggatgt atgcgtccgt
781 cttgccctgt ggctcctctg gggactcctc ctgcaccagg gccagagcct cagccatagt
841 cacagtgaga aggcgacagg aaccagctcg ggtaccatgg tgagcaaggg cgaggagctg
901 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
961 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc
1021 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc
1081 gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
1141 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
1201 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
1261 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc
1321 cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc
1381 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc
1441 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg
1501 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
1561 gggatcactc tcggcatgga cgagctgtac aagggcggcc gcagtggtgg gctggataca
1621 gtgctctttg ggcctcctct cttgactcgc cataatcacc tcaaggactt catgctggtg
1681 gtgtctatcg ttattggtgt gggcggctgc tggtttgcct atatccagaa ccgttactcc
1741 aaggagcacg gagcgggcgc tggtgccgga gctggagcag gtgctctcga gttggctact
1801 acacttgaac gtattgagaa gaactttgtc attactgacc caagattgcc agataatccc
1861 attatattcg cgtccgatag tttcttgcag ttgacagaat atagccgtga agaaattttg
1921 ggaagaaact gcaggtttct acaaggtcct gaaactgatc gcgcgacagt gagaaaaatt
1981 agagatgcca tagataacca aacagaggtc actgttcagc tgattaatta tacaaagagt
2041 ggtaaaaagt tctggaacct ctttcacttg cagcctatgc gagatcagaa gggagatgtc
2101 cagtacttta ttggggttca gttggatgga actgagcatg tccgagatgc tgccgagaga
2161 gagggagtca tgctgattaa gaaaactgca gaaaatattg atgaggcggc aaaagaactt
2221 agaattctaa tggagaaaaa atctaagccc aaaaacagtg tatggaagag gctaaaatca
2281 ccattccgga agaagaaaga ttcagtaact tgataaggat cctctagagg gcccgtttaa
2341 acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc
2401 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag
2461 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag
2521 gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct
2581 atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt
2641 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
2701 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
2761 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
2821 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
2881 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
2941 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
3001 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa
3061 ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
3121 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
3181 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
3241 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
3301 gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga
3361 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta
3421 tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag
3481 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
3541 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
3601 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag
3661 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
3721 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
3781 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
3841 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
3901 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
3961 tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg
4021 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
4081 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
4141 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
4201 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
4261 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
4321 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
4381 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt
4441 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
4501 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
4561 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
4621 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
4681 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
4741 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
4801 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
4861 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
4921 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
4981 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
5041 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
5101 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
5161 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
5221 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
5281 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
5341 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
5401 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
5461 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
5521 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
5581 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
5641 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
5701 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
5761 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
5821 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
5881 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
5941 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
6001 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
6061 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
6121 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
6181 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
6241 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
6301 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
6361 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
6421 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
6481 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
6541 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
6601 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
6661 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
6721 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct
6781 cccgatcccc tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
6841 tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct
6901 acaacaa
//
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