| 质粒名称: | APEX2 - Actin in pEGFP |
|---|---|
| 出品公司: | Clontech |
| 目录编号: | 66172 |
| 存储实验室: | Alice Ting |
|
载体骨架基本信息 |
|
| 载体名称: | pEGFP |
| 载体出品公司: | Clontech |
| 空载体大小: | 3954 bp |
| 完整质粒大小: | 5907 bp |
| 载体类型: | 哺乳细胞表达 |
| 载体抗性: | Kanamycin |
| 生长温度: | 37 ℃ |
| 宿主菌: | XL1 Blue |
| 拷贝数: | 未知 |
|
插入基因信息 |
|
| 插入基因名称: | APEX2 - human beta Actin |
| 物种来源: | H. sapiens (human), Synthetic |
| 插入大小: | 1953bp |
| 基因突变: | APEX2 contains 4 mutations relative to wt soybean APX: K14D, W41F, E112K, A134P |
| 启动子: | CMV |
| 标签/融合蛋白: | Flag epitope tag (DYKDDDDK) (N terminal on insert) |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | NheI(未破坏) |
| 3’克隆位点: | BamHI(未破坏) |
| 5’测序引物 | CMVf (CGCAAATGGGCGGTAGGCGTG) |
| 3’测序引物 | SV40pA-R |
|
文献引用方法 |
|
| 材料和方法部分: | APEX2 - Actin in pEGFP was a gift from Alice Ting (Addgene plasmid # 66172 ; http://n2t.net/addgene:66172 ; RRID:Addgene_66172) |
| 参考文献部分: |
Directed evolution of APEX2 for electron microscopy and proximity labeling. Lam SS, Martell JD, Kamer KJ, Deerinck TJ, Ellisman MH, Mootha VK, Ting AY. Nat Methods. 2014 Nov 24. doi: 10.1038/nmeth.3179. 10.1038/nmeth.3179 PubMed 25419960
|
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 APEX2 - Actin in pEGFP 5907 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION APEX2 fused to the N-terminus of human beta actin.
ACCESSION .
VERSION .
KEYWORDS APEX2 - Actin in pEGFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5907)
AUTHORS Lam SS, Martell JD, Kamer KJ, Deerinck TJ, Ellisman MH, Mootha VK,
Ting AY
TITLE Directed evolution of APEX2 for electron microscopy and proximity
labeling.
JOURNAL Nat Methods. 2014 Nov 24. doi: 10.1038/nmeth.3179.
PUBMED 25419960
REFERENCE 2 (bases 1 to 5907)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..5907
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 119..422
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 423..626
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 577..597
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
regulatory 665..674
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 674..697
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
polyA_signal 2753..2874
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2790..2809)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 2844..2863
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(2881..3336)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3018..3039)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 3230..3249
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 3363..3467
/gene="bla"
/label=AmpR promoter
promoter 3469..3826
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(3492..3512)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 3677..3812
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3739..3758
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3861..4655
/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(3915..3934)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 4525..4544
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(4843..4862)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 4887..4934
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 5263..5851
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5752..5771
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcatta
61 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
121 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
181 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
241 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
301 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
361 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
421 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
481 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
541 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
601 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc tagcgctacc
661 ggtcgccacc atggactaca aggatgacga cgataagggg aaatcatacc caacagtgtc
721 cgcagactac caggatgccg tggagaaagc caagaagaag ctgagagggt ttatcgcaga
781 aaagaggtgc gcacctctga tgctgagact ggctttccac agcgcaggca cctttgacaa
841 gggaaccaaa acaggcggac cctttggaac aatcaagcac cctgctgaac tggcacatag
901 tgccaacaat gggctggaca tcgccgtgcg gctgctggaa cctctgaaag cagagttccc
961 aattctgtcc tacgccgatt tttatcagct ggcaggagtg gtcgctgtgg aggtcactgg
1021 gggccccaag gtgcctttcc acccaggacg ggaggacaaa ccagaaccac ctccagaggg
1081 gcgcctgcca gatccgacaa agggctccga ccatctgcga gatgtgtttg ggaaagctat
1141 gggcctgact gaccaggata tcgtcgcact gtctggaggg cacaccattg gcgccgctca
1201 taaggaaagg tcaggcttcg agggaccctg gacaagcaac cccctgattt tcgacaattc
1261 ttactttact gaactgctga gtggagagaa ggaagggctg ctgcagctgc ccagcgacaa
1321 agccctgctg tccgatcccg tgttcagacc tctggtcgat aagtatgcag ccgacgagga
1381 tgcttttttc gcagattacg cagaagcaca tcagaagctg tcagaactgg gatttgccga
1441 cgccaagggc tcgggctcga cctcgggctc gggctccgga ctcagatctc gagctatgga
1501 tgatgatatc gccgcgctcg tcgtcgacaa cggctccggc atgtgcaagg ccggcttcgc
1561 gggcgacgat gccccccggg ccgtcttccc ctccatcgtg gggcgcccca ggcaccaggg
1621 cgtgatggtg ggcatgggtc agaaggattc ctatgtgggc gacgaggccc agagcaagag
1681 aggcatcctc accctgaagt accccatcga gcacggcatc gtcaccaact gggacgacat
1741 ggagaaaatc tggcaccaca ccttctacaa tgagctgcgt gtggctcccg aggagcaccc
1801 cgtgctgctg accgaggccc ccctgaaccc caaggccaac cgcgagaaga tgacccagat
1861 catgtttgag accttcaaca ccccagccat gtacgttgct atccaggctg tgctatccct
1921 gtacgcctct ggccgtacca ctggcatcgt gatggactcc ggtgacgggg tcacccacac
1981 tgtgcccatc tacgaggggt atgccctccc ccatgccatc ctgcgtctgg acctggctgg
2041 ccgggacctg actgactacc tcatgaagat cctcaccgag cgcggctaca gcttcaccac
2101 cacggccgag cgggaaatcg tgcgtgacat taaggagaag ctgtgctacg tcgccctgga
2161 cttcgagcaa gagatggcca cggctgcttc cagctcctcc ctggagaaga gctacgagct
2221 gcctgacggc caggtcatca ccattggcaa tgagcggttc cgctgccctg aggcactctt
2281 ccagccttcc ttcctgggca tggagtcctg tggcatccac gaaactacct tcaactccat
2341 catgaagtgt gacgtggaca tccgcaaaga cctgtacgcc aacacagtgc tgtctggcgg
2401 caccaccatg taccctggca ttgccgacag gatgcagaag gagatcactg ccctggcacc
2461 cagcacaatg aagatcaaga tcattgctcc tcctgagcgc aagtactccg tgtggatcgg
2521 cggctccatc ctggcctcgc tgtccacctt ccagcagatg tggatcagca agcaggagta
2581 tgacgagtcc ggcccctcca tcgtccaccg caaatgcttc tagggatcca ccggatctag
2641 ataactgatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct
2701 cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt
2761 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
2821 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaacgcgt
2881 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
2941 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
3001 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
3061 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
3121 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
3181 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
3241 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
3301 cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg
3361 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
3421 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa
3481 gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
3541 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
3601 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
3661 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
3721 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
3781 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga
3841 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
3901 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
3961 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
4021 ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg
4081 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
4141 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
4201 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
4261 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
4321 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
4381 tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
4441 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
4501 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
4561 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
4621 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
4681 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
4741 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
4801 tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac
4861 aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt
4921 tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc
4981 accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc
5041 aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc
5101 ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
5161 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
5221 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
5281 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
5341 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
5401 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
5461 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
5521 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
5581 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
5641 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
5701 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
5761 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
5821 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
5881 cctggccttt tgctggcctt ttgctca
//
