| 质粒名称: | APEX2 - Tubulin in pEGFP |
|---|---|
| 出品公司: | Clontech |
| 目录编号: | 66171 |
| 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: | 3954bp |
| 完整质粒大小: | 6132 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: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | APEX2 - alpha tubulin |
| 物种来源: | Synthetic |
| 插入大小: | 2178 bp |
| 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) |
|
文献引用方法 |
|
| 材料和方法部分: | APEX2 - Tubulin in pEGFP was a gift from Alice Ting (Addgene plasmid # 66171 ; http://n2t.net/addgene:66171 ; RRID:Addgene_66171) |
| 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 - Tubulin in pEGFP 6132 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION APEX2 fused to the N-terminus of alpha tubulin.
ACCESSION .
VERSION .
KEYWORDS APEX2 - Tubulin in pEGFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6132)
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 6132)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..6132
/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 616..639
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
polyA_signal 2920..3041
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2957..2976)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3011..3030
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(3048..3503)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3185..3206)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 3397..3416
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 3530..3634
/gene="bla"
/label=AmpR promoter
promoter 3636..3993
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(3659..3679)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 3844..3979
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3906..3925
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 4028..4822
/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(4082..4101)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 4692..4711
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(5010..5029)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 5054..5101
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 5430..6018
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5919..5938
/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 ccatggacta caaggatgac gacgataagg ggaaatcata cccaacagtg
661 tccgcagact accaggatgc cgtggagaaa gccaagaaga agctgagagg gtttatcgca
721 gaaaagaggt gcgcacctct gatgctgaga ctggctttcc acagcgcagg cacctttgac
781 aagggaacca aaacaggcgg accctttgga acaatcaagc accctgctga actggcacat
841 agtgccaaca atgggctgga catcgccgtg cggctgctgg aacctctgaa agcagagttc
901 ccaattctgt cctacgccga tttttatcag ctggcaggag tggtcgctgt ggaggtcact
961 gggggcccca aggtgccttt ccacccagga cgggaggaca aaccagaacc acctccagag
1021 gggcgcctgc cagatccgac aaagggctcc gaccatctgc gagatgtgtt tgggaaagct
1081 atgggcctga ctgaccagga tatcgtcgca ctgtctggag ggcacaccat tggcgccgct
1141 cataaggaaa ggtcaggctt cgagggaccc tggacaagca accccctgat tttcgacaat
1201 tcttacttta ctgaactgct gagtggagag aaggaagggc tgctgcagct gcccagcgac
1261 aaagccctgc tgtccgatcc cgtgttcaga cctctggtcg ataagtatgc agccgacgag
1321 gatgcttttt tcgcagatta cgcagaagca catcagaagc tgtcagaact gggatttgcc
1381 gacgccaagg gctcgggctc gacctcgggc tcgggctccg gactcagatc tcgagtgcgt
1441 gagtgcatct ccatccacgt tggccaggct ggtgtccaga ttggcaatgc ctgctgggag
1501 ctctactgcc tggaacacgg catccagccc gatggccaga tgccaagtga caagaccatt
1561 gggggaggag atgactcctt caacaccttc ttcagtgaga cgggcgctgg caagcacgtg
1621 ccccgggctg tgtttgtaga cttggaaccc acagtcattg atgaagttcg cactggcacc
1681 taccgccagc tcttccaccc tgagcagctc atcacaggca aggaagatgc tgccaataac
1741 tatgcccgag ggcactacac cattggcaag gagatcattg accttgtgtt ggaccgaatt
1801 cgcaagctgg ctgaccagtg caccggtctt cagggcttct tggttttcca cagctttggt
1861 gggggaactg gttctgggtt cacctccctg ctcatggaac gtctctcagt tgattatggc
1921 aagaagtcca agctggagtt ctccatttac ccagcacccc aggtttccac agctgtagtt
1981 gagccctaca actccatcct caccacccac accaccctgg agcactctga ttgtgccttc
2041 atggtagaca atgaggccat ctatgacatc tgtcgtagaa acctcgatat cgagcgccca
2101 acctacacta accttaaccg ccttattagc cagattgtgt cctccatcac tgcttccctg
2161 agatttgatg gagccctgaa tgttgacctg acagaattcc agaccaacct ggtgccctac
2221 ccccgcatcc acttccctct ggccacatat gcccctgtca tctctgctga gaaagcctac
2281 catgaacagc tttctgtagc agagatcacc aatgcttgct ttgagccagc caaccagatg
2341 gtgaaatgtg accctcgcca tggtaaatac atggcttgct gcctgttgta ccgtggtgac
2401 gtggttccca aagatgtcaa tgctgccatt gccaccatca aaaccaagcg cagcatccag
2461 tttgtggatt ggtgccccac tggcttcaag gttggcatca actaccagcc tcccactgtg
2521 gtgcctggtg gagacctggc caaggtacag agagctgtgt gcatgctgag caacaccaca
2581 gccattgctg aggcctgggc tcgcctggac cacaagtttg acctgatgta tgccaagcgt
2641 gcctttgttc actggtacgt gggtgagggg atggaggaag gcgagttttc agaggcccgt
2701 gaagatatgg ctgcccttga gaaggattat gaggaggttg gtgtggattc tgttgaagga
2761 gagggtgagg aagaaggaga ggaatactaa ggatccaccg gatctagata actgatcata
2821 atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc
2881 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat
2941 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
3001 cattctagtt gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa ttgtaagcgt
3061 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
3121 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
3181 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
3241 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
3301 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
3361 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
3421 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
3481 taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
3541 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
3601 ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg
3661 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
3721 agcatgcatc tcaattagtc agcaaccagg kgyggaaagt ccccaggctc cccagcaggc
3781 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
3841 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
3901 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
3961 ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc
4021 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
4081 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
4141 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
4201 tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
4261 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
4321 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
4381 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
4441 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
4501 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat
4561 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
4621 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
4681 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
4741 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
4801 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
4861 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
4921 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
4981 ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat accggaagga
5041 acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt
5101 cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat
5161 tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa
5221 ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca
5281 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
5341 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
5401 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
5461 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
5521 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
5581 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
5641 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
5701 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
5761 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
5821 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
5881 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
5941 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
6001 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
6061 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
6121 accgccatgc at
//
