| 质粒名称: | pHaloTag-EGFP |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 86629 |
| 存储实验室: | Thomas Leonard, Ivan Yudushkin |
|
载体骨架基本信息 |
|
| 载体名称: | pEGFP-N1 |
| 载体出品公司: | -- |
| 载体抗性: | 卡那霉素 |
| 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: | -- |
| 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: | HaloTag,Promega HaloTag |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | -- |
| 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: | pHaloTag-EGFP was a gift from Thomas Leonard & Ivan Yudushkin (Addgene plasmid # 86629 ; http://n2t.net/addgene:86629 ; RRID:Addgene_86629) |
| 参考文献部分: | PI(3,4,5)P3 Engagement Restricts Akt Activity to Cellular Membranes. Ebner M, Lucic I, Leonard TA, Yudushkin I. Mol Cell. 2017 Feb 2;65(3):416-431.e6. doi: 10.1016/j.molcel.2016.12.028. 10.1016/j.molcel.2016.12.028 PubMed 28157504 |
买家导航
pHaloTagEGFP载体基本信息
pHaloTagEGFP载体图谱质粒图谱和多克隆位点信息
pHaloTagEGFP载体简介
pHaloTagEGFP载体序列
LOCUS pHaloTag-EGFP 5602 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Expresses HaloTag-eGFP tandem (cytosolic).
ACCESSION .
VERSION .
KEYWORDS pHaloTag-EGFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5602)
AUTHORS Ebner M, Lucic I, Leonard TA, Yudushkin I
TITLE PI(3,4,5)P3 Engagement Restricts Akt Activity to Cellular Membranes.
JOURNAL Mol Cell. 2017 Feb 2;65(3):416-431.e6. doi:
10.1016/j.molcel.2016.12.028.
PUBMED 28157504
REFERENCE 2 (bases 1 to 5602)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5602
/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 600..609
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 606..1496
/codon_start=1
/product="modified bacterial dehalogenase that forms
covalent bonds with chloroalkane derivatives"
/label=HaloTag(R)
/translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
IGSEIARWLSTLEISG"
CDS 1551..2267
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
FVTAAGITLGMDELYK"
primer_bind complement(1593..1614)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(1854..1873)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 2201..2222
/label=EGFP-C
/note="EGFP, forward primer"
polyA_signal 2390..2511
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2427..2446)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 2481..2500
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(2518..2973)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2655..2676)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2867..2886
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 3000..3104
/gene="bla"
/label=AmpR promoter
promoter 3106..3463
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(3129..3149)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 3314..3449
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3376..3395
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3498..4292
/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(3552..3571)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 4162..4181
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(4480..4499)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 4524..4571
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 4900..5488
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5389..5408
/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 gctagcaaag
601 ccaccatggc agaaatcggt actggctttc cattcgaccc ccattatgtg gaagtcctgg
661 gcgagcgcat gcactacgtc gatgttggtc cgcgcgatgg cacccctgtg ctgttcctgc
721 acggtaaccc gacctcctcc tacgtgtggc gcaacatcat cccgcatgtt gcaccgaccc
781 atcgctgcat tgctccagac ctgatcggta tgggcaaatc cgacaaacca gacctgggtt
841 atttcttcga cgaccacgtc cgcttcatgg atgccttcat cgaagccctg ggtctggaag
901 aggtcgtcct ggtcattcac gactggggct ccgctctggg tttccactgg gccaagcgca
961 atccagagcg cgtcaaaggt attgcattta tggagttcat ccgccctatc ccgacctggg
1021 acgaatggcc agaatttgcc cgcgagacct tccaggcctt ccgcaccacc gacgtcggcc
1081 gcaagctgat catcgatcag aacgttttta tcgagggtac gctgccgatg ggtgtcgtcc
1141 gcccgctgac tgaagtcgag atggaccatt accgcgagcc gttcctgaat cctgttgacc
1201 gcgagccact gtggcgcttc ccaaacgagc tgccaatcgc cggtgagcca gcgaacatcg
1261 tcgcgctggt cgaagaatac atggactggc tgcaccagtc ccctgtcccg aagctgctgt
1321 tctggggcac cccaggcgtt ctgatcccac cggccgaagc cgctcgcctg gccaaaagcc
1381 tgcctaactg caaggctgtg gacatcggcc cgggtctgaa tctgctgcaa gaagacaacc
1441 cggacctgat cggcagcgag atcgcgcgct ggctgtcgac gctcgagatt tccggccacc
1501 ggtcgcaacc ttgggtaccg cgggcccggg atccaccggt cgcaaccttg gtgagcaagg
1561 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
1621 gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc
1681 tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc
1741 tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct
1801 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg
1861 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
1921 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
1981 actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga
2041 acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc
2101 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc
2161 agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
2221 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaaagc ggccgcgact
2281 ctagatcata atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc
2341 acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat
2401 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
2461 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa
2521 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
2581 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
2641 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
2701 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
2761 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
2821 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
2881 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
2941 ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc
3001 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
3061 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa
3121 ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
3181 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
3241 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
3301 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
3361 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
3421 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag
3481 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
3541 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
3601 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
3661 gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg
3721 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
3781 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
3841 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
3901 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
3961 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
4021 aggcgagcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
4081 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
4141 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
4201 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
4261 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
4321 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
4381 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
4441 catgctggag ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat
4501 accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg
4561 gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc
4621 gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag
4681 ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc
4741 aggttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
4801 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
4861 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
4921 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
4981 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
5041 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
5101 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
5161 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
5221 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
5281 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
5341 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
5401 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
5461 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
5521 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
5581 taaccgtatt accgccatgc at
//
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