| 质粒名称: | GFP-2xPH-TAPP1 WT |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 161990 |
| 存储实验室: | David Bryant |
|
载体骨架基本信息 |
|
| 载体名称: | eGFP-C1 |
| 载体出品公司: | -- |
| 空载体大小: | -- |
| 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） |
| 载体抗性: | 卡那霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | NEB Stable |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | PH-TAPP1,PLEKHA1 |
| 物种来源: | H. sapiens (human) |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子: | -- |
| 标签/融合蛋白: | eGFP（主干上的N端） |
| 克隆方法: | Ligation Independent克隆 |
| 5’克隆位点: | -- |
| 3’克隆位点: | -- |
| 5’测序引物 | -- |
| 3’测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | GFP-2xPH-TAPP1 WT was a gift from David Bryant (Addgene plasmid # 161990 ; http://n2t.net/addgene:161990 ; RRID:Addgene_161990) |
| 参考文献部分: | The phospholipid PI(3,4)P2 is an apical identity determinant. Roman-Fernandez A, Roignot J, Sandilands E, Nacke M, Mansour MA, McGarry L, Shanks E, Mostov KE, Bryant DM. Nat Commun. 2018 Nov 28;9(1):5041. doi: 10.1038/s41467-018-07464-8. 10.1038/s41467-018-07464-8 PubMed 30487552 |
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-2xPH-TAPP1 WT 6747 bp ds-DNA circular SYN 13-MAY-2021
DEFINITION Expresses two tandem repeats of Tapp1 PH domain that binds to
PI(3,4)P2. Fused to eGFP.
ACCESSION .
VERSION .
KEYWORDS GFP-2xPH-TAPP1 WT
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6747)
AUTHORS Roman-Fernandez A, Roignot J, Sandilands E, Nacke M, Mansour MA,
McGarry L, Shanks E, Mostov KE, Bryant DM
TITLE The phospholipid PI(3,4)P2 is an apical identity determinant.
JOURNAL Nat Commun. 2018 Nov 28;9(1):5041. doi: 10.1038/s41467-018-07464-8.
PUBMED 30487552
REFERENCE 2 (bases 1 to 6747)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6747
/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"
regulatory 2302..2311
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 2311..2340
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
polyA_signal 3535..3656
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3572..3591)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3626..3645
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(3663..4118)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3800..3821)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 4012..4031
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 4145..4249
/gene="bla"
/label=AmpR promoter
promoter 4251..4608
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(4274..4294)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 4459..4594
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 4521..4540
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 4643..5437
/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(4697..4716)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 5307..5326
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(5625..5644)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 5669..5716
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 6045..6633
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6534..6553
/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 atctcgagtg aatgatcagc aggacctagt ggaatgggta
1381 aatgtgttaa acaaagctat aaaaattaca gtaccaaagc agtcagactc acagcctaat
1441 tctgataacc taagtcgcca tggtgaatgt gggaaaaagc aagtgtctta cagaactgat
1501 attgttggtg gcgtacccat cattactccc actcagaaag aagaagtaaa tgaatgtggt
1561 gaaagtattg acagaaataa tctgaaacgg tcacaaagcc atcttcctta ctttactcct
1621 aaaccacctc aagatagtgc ggttatcaaa gctggatatt gtgtaaaaca aggagcagtg
1681 atgaaaaact ggaagagaag atattttcaa ttggatgaaa acacaatagg ctacttcaaa
1741 tctgaactgg aaaaggaacc tcttcgcgta ataccactta aagaggttca taaagtccag
1801 gaatgtaagc aaagcgacat aatgatgagg gacaacctct ttgaaattgt aacaacgtct
1861 cgaactttct atgtgcaggc tgatagccct gaagagatgc acagttggat taaagcagtc
1921 tctggcgcca ttgtagcaca gcggggtccc ggcagatctg cgtcttctga gcatcccccc
1981 ggtccttcag aatccaaaca cgctttccgt cctaccaacg cagccgccgc cacctcacat
2041 tccacagcct ctcgcagcaa ctctttggtc tcaaccttta ccatggagaa gcgaggattt
2101 tacgagtctc ttgccaaggt caagccaggg aacttcaagg tccagactgt ctctccaaga
2161 gaaccagctt ccaaagtgac tgaacaagct ctgttaagac ctcaaagtaa aaatggccct
2221 caggaaaaag attgtgacct agtagacttg gacgatgcga gccttccggt cagtgacgtg
2281 ggtaccgcgg gcccgggatc cgccaccatg gaacaaaaac tcatctcaga agaggatctg
2341 aatgatcagc aggacctagt ggaatgggta aatgtgttaa acaaagctat aaaaattaca
2401 gtaccaaagc agtcagactc acagcctaat tctgataacc taagtcgcca tggtgaatgt
2461 gggaaaaagc aagtgtctta cagaactgat attgttggtg gcgtacccat cattactccc
2521 actcagaaag aagaagtaaa tgaatgtggt gaaagtattg acagaaataa tctgaaacgg
2581 tcacaaagcc atcttcctta ctttactcct aaaccacctc aagatagtgc ggttatcaaa
2641 gctggatatt gtgtaaaaca aggagcagtg atgaaaaact ggaagagaag atattttcaa
2701 ttggatgaaa acacaatagg ctacttcaaa tctgaactgg aaaaggaacc tcttcgcgta
2761 ataccactta aagaggttca taaagtccag gaatgtaagc aaagcgacat aatgatgagg
2821 gacaacctct ttgaaattgt aacaacgtct cgaactttct atgtgcaggc tgatagccct
2881 gaagagatgc acagttggat taaagcagtc tctggcgcca ttgtagcaca gcggggtccc
2941 ggcagatctg cgtcttctga gcatcccccc ggtccttcag aatccaaaca cgctttccgt
3001 cctaccaacg cagccgccgc cacctcacat tccacagcct ctcgcagcaa ctctttggtc
3061 tcaaccttta ccatggagaa gcgaggattt tacgagtctc ttgccaaggt caagccaggg
3121 aacttcaagg tccagactgt ctctccaaga gaaccagctt ccaaagtgac tgaacaagct
3181 ctgttaagac ctcaaagtaa aaatggccct caggaaaaag attgtgacct agtagacttg
3241 gacgatgcga gccttccggt cagtgacgtg tgaggcagaa gcgcacggag cctgcctgcc
3301 tctgccgtcc tcagttacct ttcatgaggc ttctagccaa agatgataaa gggggaaatg
3361 gtttttagtg cgtatattat actgcctctt aggtgtactc tttgaggatc caccggatct
3421 agataactga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac
3481 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
3541 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
3601 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaacgc
3661 gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
3721 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
3781 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
3841 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
3901 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
3961 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
4021 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
4081 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa
4141 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
4201 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga
4261 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
4321 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
4381 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
4441 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
4501 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
4561 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga
4621 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
4681 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
4741 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
4801 gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac
4861 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
4921 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
4981 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
5041 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
5101 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
5161 gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
5221 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
5281 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
5341 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
5401 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
5461 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
5521 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
5581 gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag
5641 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt
5701 gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc
5761 ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc
5821 ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata
5881 gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
5941 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
6001 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
6061 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
6121 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
6181 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
6241 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
6301 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
6361 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
6421 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
6481 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
6541 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
6601 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
6661 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
6721 gtggataacc gtattaccgc catgcat
//
