| 质粒名称: | pJJiaDEST-APEX2-GAL9 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 149520 |
| 存储实验室: | Vojo Deretic |
|
载体骨架基本信息 |
|
| 载体名称: | pJJiaDEST-APEX2 |
| 载体出品公司: | -- |
| 空载体大小: | -- |
| 完整质粒大小: | 7225 bp |
| 载体修饰: | -- |
| 载体类型: | 哺乳动物表达 |
| 筛选标记: | -- |
| 载体抗性: | 氨苄青霉素，100μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | LGALS9 |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 972 bp |
| 基因突变: | -- |
| GenBank编号 | NP_002299.2 |
| 启动子: | CMV |
| 标签/融合蛋白: | APEX2（插入件上的N端子） |
| 克隆方法: | 网关克隆 |
| 5’克隆位点: | -- |
| 3’克隆位点: | -- |
| 5’测序引物 | CMV |
| 3’测序引物 | BGHR |
|
文献引用方法 |
|
| 材料和方法部分: | pJJiaDEST-APEX2-GAL9 was a gift from Vojo Deretic (Addgene plasmid # 149520 ; http://n2t.net/addgene:149520 ; RRID:Addgene_149520) |
| 参考文献部分: | Galectins Control mTOR in Response to Endomembrane Damage. Jia J, Abudu YP, Claude-Taupin A, Gu Y, Kumar S, Choi SW, Peters R, Mudd MH, Allers L, Salemi M, Phinney B, Johansen T, Deretic V. Mol Cell. 2018 Apr 5;70(1):120-135.e8. doi: 10.1016/j.molcel.2018.03.009. 10.1016/j.molcel.2018.03.009 PubMed 29625033 |
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 pJJiaDEST-APEX2-GAL9 7225 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Express APEX2-Galectin-9 in mammalian cells.
ACCESSION .
VERSION .
KEYWORDS pJJiaDEST-APEX2-GAL9
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7225)
AUTHORS Jia J, Abudu YP, Claude-Taupin A, Gu Y, Kumar S, Choi SW, Peters R,
Mudd MH, Allers L, Salemi M, Phinney B, Johansen T, Deretic V
TITLE Galectins Control mTOR in Response to Endomembrane Damage.
JOURNAL Mol Cell. 2018 Apr 5;70(1):120-135.e8. doi:
10.1016/j.molcel.2018.03.009.
PUBMED 29625033
REFERENCE 2 (bases 1 to 7225)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..7225
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(44..63)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 235..614
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 615..818
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 769..789
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 863..882
/label=T7
/note="T7 promoter, forward primer"
promoter 863..881
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
regulatory 901..910
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 910..933
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
CDS 934..1680
/codon_start=1
/product="soybean ascorbate peroxidase, engineered to serve
as a monomeric tag for electron microscopy and live-cell
proteomics (Lam et al., 2015)"
/label=APEX2
/note="more active than the original APEX due to the A134P
mutation"
/translation="GKSYPTVSADYQDAVEKAKKKLRGFIAEKRCAPLMLRLAFHSAGT
FDKGTKTGGPFGTIKHPAELAHSANNGLDIAVRLLEPLKAEFPILSYADFYQLAGVVAV
EVTGGPKVPFHPGREDKPEPPPEGRLPDPTKGSDHLRDVFGKAMGLTDQDIVALSGGHT
IGAAHKERSGFEGPWTSNPLIFDNSYFTELLSGEKEGLLQLPSDKALLSDPVFRPLVDK
YAADEDAFFADYAEAHQKLSELGFADA"
CDS 1687..1713
/codon_start=1
/product="nuclear export signal from the HIV Rev protein
(Fischer et al., 1995)"
/label=NES
/translation="LPPLERLTL"
protein_bind 1726..1750
/gene="mutant version of attB"
/label=attB1
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
protein_bind complement(2723..2747)
/gene="mutant version of attB"
/label=attB2
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
promoter complement(2778..2796)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(2779..2796)
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind complement(2816..2833)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 2822..3046
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 3092..3520
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3179..3198)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 3389..3410
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(3529..3549)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 3534..3863
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 3714..3849
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3776..3795
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3930..4724
/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
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(3984..4003)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 4594..4613
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 4898..5019
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4935..4954)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4989..5008
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(5068..5084)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5068..5084)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(5081..5103)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 5092..5108
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(5116..5146)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5161..5182
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(5299..5316)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5470..6058)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5550..5569)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(6229..7089)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 6852..6871
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(7090..7194)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttgcggcc
901 gccaccatgg actacaagga tgacgacgat aagggaaagt cttacccaac tgtgagtgct
961 gattaccagg acgccgttga gaaggcgaag aagaagctca gaggcttcat cgctgagaag
1021 agatgcgctc ctctaatgct ccgtttggca ttccactctg ctggaacctt tgacaagggc
1081 acgaagaccg gtggaccctt cggaaccatc aagcaccctg ccgaactggc tcacagcgct
1141 aacaacggtc ttgacatcgc tgttaggctt ttggagccac tcaaggcgga gttccctatt
1201 ttgagctacg ccgatttcta ccagttggct ggcgttgttg ccgttgaggt cacgggtgga
1261 cctaaggttc cattccaccc tggaagagag gacaagcctg agccaccacc agagggtcgc
1321 ttgcccgatc ccactaaggg ttctgaccat ttgagagatg tgtttggcaa agctatgggg
1381 cttactgacc aagatatcgt tgctctatct gggggtcaca ctattggagc tgcacacaag
1441 gagcgttctg gatttgaggg tccctggacc tctaatcctc ttattttcga caactcatac
1501 ttcacggagt tgttgagtgg tgagaaggaa ggtctccttc agctaccttc tgacaaggct
1561 cttttgtctg accctgtatt ccgccctctc gttgacaaat atgcagcgga cgaagatgcc
1621 ttctttgctg attacgctga ggctcaccaa aagctttccg agcttgggtt tgctgatgcc
1681 ctgcagctgc ctcccctgga gcgcctgacc ctggacctcg aatcaacaag tttgtacaaa
1741 aaagcaggct tcgccttcag cggttcccag gctccctacc tgagtccagc tgtccccttt
1801 tctgggacta ttcaaggagg tctccaggac ggacttcaga tcactgtcaa tgggaccgtt
1861 ctcagctcca gtggaaccag gtttgctgtg aactttcaga ctggcttcag tggaaatgac
1921 attgccttcc acttcaaccc tcggtttgaa gatggagggt acgtggtgtg caacacgagg
1981 cagaacggaa gctgggggcc cgaggagagg aagacacaca tgcctttcca gaaggggatg
2041 ccctttgacc tctgcttcct ggtgcagagc tcagatttca aggtgatggt gaacgggatc
2101 ctcttcgtgc agtacttcca ccgcgtgccc ttccaccgtg tggacaccat ctccgtcaat
2161 ggctctgtgc agctgtccta catcagcttc cagcctcccg gcgtgtggcc tgccaacccg
2221 gctcccatta cccagacagt catccacaca gtgcagagcg cccctggaca gatgttctct
2281 actcccgcca tcccacctat gatgtacccc caccccgcct atccgatgcc tttcatcacc
2341 accattctgg gagggctgta cccatccaag tccatcctcc tgtcaggcac tgtcctgccc
2401 agtgctcaga ggttccacat caacctgtgc tctgggaacc acatcgcctt ccacctgaac
2461 ccccgttttg atgagaatgc tgtggtccgc aacacccaga tcgacaactc ctgggggtct
2521 gaggagcgaa gtctgccccg aaaaatgccc ttcgtccgtg gccagagctt ctcagtgtgg
2581 atcttgtgtg aagctcactg cctcaaggtg gccgtggatg gtcagcacct gtttgaatac
2641 taccatcgcc tgaggaacct gcccaccatc aacagactgg aagtgggggg cgacatccag
2701 ctgacccatg tgcagacata ggacccagct ttcttgtaca aagtggttga ttcgagcatg
2761 catctagagg gccctattct atagtgtcac ctaaatgcta gagctcgctg atcagcctcg
2821 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
2881 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
2941 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
3001 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa
3061 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
3121 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
3181 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
3241 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
3301 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
3361 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
3421 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
3481 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
3541 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
3601 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
3661 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
3721 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
3781 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
3841 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg
3901 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
3961 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
4021 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
4081 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg
4141 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
4201 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
4261 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
4321 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
4381 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
4441 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
4501 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
4561 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
4621 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
4681 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
4741 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
4801 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
4861 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
4921 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
4981 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
5041 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
5101 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
5161 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
5221 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
5281 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
5341 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
5401 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
5461 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
5521 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
5581 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
5641 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
5701 cgttcgctcc aagctgggct gtgtgcacaa accccccgtt cagcccgacc gctgcgcctt
5761 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
5821 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
5881 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
5941 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
6001 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
6061 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
6121 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
6181 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
6241 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
6301 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
6361 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
6421 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
6481 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
6541 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
6601 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
6661 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
6721 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
6781 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
6841 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
6901 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
6961 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
7021 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
7081 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
7141 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
7201 tccccgaaaa gtgccacctg acgtc
//
载体图谱质粒图谱pdf版和相关资料下载
载体质粒应用举例
- 上一篇：pHAGE UbiC scAB-GFP
- 下一篇：pDGM3B
