| 质粒名称: | CFP-C1-PLCdelta-PH |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 21262 |
| 存储实验室: | Tobias Meyer |
|
载体骨架基本信息 |
|
| 载体名称: | C1 |
| 载体出品公司: | Clontech |
| 载体抗性: | 卡那霉素 |
| 空载体大小: | 4701 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 新霉素（G418） |
| 高拷贝/低拷贝: | -- |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | PLCdelta |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 528 bp |
| 基因突变: | PH domain of PLCdelta (human). Also known as PLCdelta-PH-CFP. Lab plasmid WC0026. |
| GenBank编号 | -- |
| 启动子 | CFP |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | BamH1(销毁时未知) |
| 3' 克隆位点: | XbaI(销毁时未知) |
| 5' 测序引物 | -- |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | CFP-C1-PLCdelta-PH was a gift from Tobias Meyer (Addgene plasmid # 21262 ; http://n2t.net/addgene:21262 ; RRID:Addgene_21262) |
| 参考文献部分: | Localized biphasic changes in phosphatidylinositol-4,5-bisphosphate at sites of phagocytosis. Botelho RJ, Teruel M, Dierckman R, Anderson R, Wells A, York JD, Meyer T, Grinstein S. J Cell Biol. 2000 Dec 25. 151(7):1353-68. 10.1083/jcb.151.7.1353 PubMed 11134066 |
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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 CFP-C1-PLCdelta-PH 5229 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS CFP-C1-PLCdelta-PH
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5229)
AUTHORS Botelho RJ, Teruel M, Dierckman R, Anderson R, Wells A, York JD,
Meyer T, Grinstein S
TITLE Localized biphasic changes in phosphatidylinositol-4,5-bisphosphate
at sites of phagocytosis.
JOURNAL J Cell Biol. 2000 Dec 25. 151(7):1353-68.
PUBMED 11134066
REFERENCE 2 (bases 1 to 5229)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5229
/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"
CDS 598..1314
/codon_start=1
/product="cyan variant of GFP"
/label=CFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHRFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
AHFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(643..664)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(904..923)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1251..1272
/label=EGFP-C
/note="EGFP, forward primer"
misc_feature 1315..1371
/label=MCS
/note="multiple cloning site"
polyA_signal 2017..2138
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2054..2073)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 2108..2127
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(2145..2600)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2282..2303)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2494..2513
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 2627..2731
/gene="bla"
/label=AmpR promoter
promoter 2733..3090
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(2756..2776)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 2941..3076
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3003..3022
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3125..3919
/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(3179..3198)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3789..3808
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(4107..4126)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 4151..4198
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 4527..5115
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5016..5035
/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 gctagcgatg
601 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
661 gacgtaaacg gccacaggtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
721 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
781 gtgaccaccc tgacctgggg cgtgcagtgc ttcagccgct accccgacca catgaagcag
841 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgtac catcttcttc
901 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
961 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
1021 ctggagtaca actacatcag ccacaacgtc tatatcaccg ccgacaagca gaagaacggc
1081 atcaaggccc acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac
1141 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
1201 ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg
1261 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagagatct
1321 cgagctcaag cttcgaattc tgcagtcgac ggtaccgcgg gcccgggatc catggactcg
1381 ggccgggact tcctgaccct gcacggccta caggatgatg aggatctaca ggcgctgctg
1441 aagggcagcc agctcctgaa ggtgaagtcc agctcatgga ggagagagcg cttctacaag
1501 ttgcaggagg actgcaagac catctggcag gagtcccgca aggtcatgcg gaccccggag
1561 tcccagctgt tctccatcga ggacattcag gaggtgcgaa tggggcaccg cacggagggt
1621 ctggagaagt tcgcccgtga tgtgcccgag gaccgctgct tctccattgt cttcaaggac
1681 cagcgcaata cactagacct catcgcccca tcgccagctg atgcccagca ctgggtgctg
1741 gggctgcaca agatcatcca ccactcaggc tccatggacc agcgtcagaa gctacagcac
1801 tggattcact cctgcttgcg aaaagctgac aaaaacaagg acaacaagat gagcttcaag
1861 gagctgcaga acttcctgaa ggagctcaac atccagtaat ctagataact gatcataatc
1921 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg
1981 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
2041 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
2101 tctagttgtg gtttgtccaa actcatcaat gtatcttaag gcgtaaattg taagcgttaa
2161 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
2221 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
2281 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
2341 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
2401 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
2461 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
2521 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
2581 tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
2641 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
2701 aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat
2761 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
2821 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
2881 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
2941 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
3001 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
3061 ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt
3121 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
3181 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
3241 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
3301 actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
3361 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
3421 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
3481 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
3541 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
3601 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc
3661 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
3721 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
3781 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
3841 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
3901 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
3961 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
4021 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
4081 ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc
4141 cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat
4201 aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg
4261 ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc
4321 ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat
4381 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
4441 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
4501 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
4561 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
4621 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta
4681 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
4741 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
4801 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
4861 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
4921 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
4981 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
5041 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
5101 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
5161 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
5221 gccatgcat
//
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