| 质粒名称: | pAAV-b2AR-eLOV-TEVcs-FLAG-GAL4-V5 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 104841 |
| 存储实验室: | Alice Ting |
|
载体骨架基本信息 |
|
| 载体名称: | pAAV |
| 载体出品公司: | Clontech |
| 空载体大小: | 4038 bp |
| 完整质粒大小: | 6398bp |
| 载体类型: | 哺乳细胞表达, AAV |
| 载体抗性: | 氨苄霉素 |
| 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: | b2AR-eLOV-TEVcs-FLAG-GAL4-V5 |
| 物种来源: | Synthetic |
| 启动子: | CMV |
| 克隆方法: | 未知 |
| 5’测序引物 | CMV-F |
|
文献引用方法 |
|
| 材料和方法部分: | pAAV-b2AR-eLOV-TEVcs-FLAG-GAL4-V5 was a gift from Alice Ting (Addgene plasmid # 104841 ; http://n2t.net/addgene:104841 ; RRID:Addgene_104841) |
| 参考文献部分: | Time-gated detection of protein-protein interactions with transcriptional readout. Kim MW, Wang W, Sanchez MI, Coukos R, von Zastrow M, Ting AY. Elife. 2017 Nov 30;6. doi: 10.7554/eLife.30233. 10.7554/eLife.30233 |
买家导航
pAAV-b2AR-eLOV-TEVcs-FLAG-GAL4-V5载体质粒基本信息
pAAV-b2AR-eLOV-TEVcs-FLAG-GAL4-V5质粒图谱载体图谱和pAAV-b2AR-eLOV-TEVcs-FLAG-GAL4-V5载体序列质粒序列多克隆位点信息
pAAV-b2AR-eLOV-TEVcs-FLAG-GAL4-V5质粒载体简介
pAAV-b2AR-eLOV-TEVcs-FLAG-GAL4-V5质粒序列载体序列
LOCUS pAAV-b2AR-eLOV-TEVcs-FLAG-GAL4-V5 6398 bp DNA circular SYN 15-DEC-2025
DEFINITION b2AR-SPARK TF component.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6398)
AUTHORS Kim MW, Wang W, Sanchez MI, Coukos R, von Zastrow M, Ting AY
TITLE Time-gated detection of protein-protein interactions with
transcriptional readout.
JOURNAL Elife. 2017 Nov 30;6. doi: 10.7554/eLife.30233.
PUBMED 29189201
REFERENCE 2 (bases 1 to 6398)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Dec 15, 2025 from SnapGene Server 8.0.1
https://www.biofeng.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Elife.";
date: "2017-11-30"; volume: "6. doi"; pages: " 10.7554/eLife.30233"
COMMENT Sequence Label: pAAV-b2AR-eLOV-TEVcs-FLAG-GAL4-V5
FEATURES Location/Qualifiers
source 1..6398
/mol_type="other DNA"
/organism="synthetic DNA construct"
repeat_region 1..130
/label=AAV2 ITR (alternate)
/note="Functional equivalent of wild-type AAV2 ITR"
enhancer 201..504
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 505..708
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 659..679
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 753..772
/label=T7
/note="T7 promoter, forward primer"
promoter 753..771
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
regulatory 785..794
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 2507..2527
/codon_start=1
/product="tobacco etch virus (TEV) protease recognition and
cleavage site"
/label=TEV site
/translation="ENLYFQM"
CDS 2540..2563
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
CDS 2564..3001
/codon_start=1
/gene="Saccharomyces cerevisiae GAL4 (truncated)"
/product="DNA binding domain of the GAL4 transcriptional
activator"
/label=GAL4 DNA binding domain
/translation="KLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTKR
SPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNKD
AVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"
CDS 3002..3343
/codon_start=1
/gene="Saccharomyces cerevisiae GAL4 (truncated)"
/product="activation domain of the GAL4 transcriptional
activator"
/label=GAL4 activation domain
/translation="ANFNQSGNIADSSLSFTFTNSSNGPNLITTQTNSQALSQPIASSN
VHDNFMNNEITASKIDDGNNSKPLSPGWTDQTAYNAFGITTGMFNTTTMDDVYNYLFDD
EDTPPNPKKE"
CDS 3344..3385
/codon_start=1
/product="epitope tag from simian virus 5"
/label=V5 tag
/translation="GKPIPNPLLGLDST"
primer_bind complement(3419..3436)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 3425..3632
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
repeat_region 3661..3801
/label=AAV2 ITR
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
repeat_region 3661..3790
/label=AAV2 ITR
rep_origin 3876..4331
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3963..3982)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 4173..4194
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(4348..4367)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 4467..4489
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(4527..4545)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 4613..4717
/gene="bla"
/label=AmpR promoter
CDS join(4718..4786,4787..5578)
/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 complement(4936..4955)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 5749..6337
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6238..6257
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
121 aggggttcct gcggccgcac gcgttgtaca tagtaatcaa ttacggggtc attagttcat
181 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
241 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
301 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
361 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
421 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
481 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
541 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
601 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
661 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact
721 agagaaccca ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa
781 gcttgccacc atggggcaac ccgggaacgg cagcgccttc ttgctggcac ccaatagaag
841 ccatgcgccg gaccacgacg tcacgcagca gagggacgag gtgtgggtgg tgggcatggg
901 catcgtcatg tctctcatcg tcctggccat cgtgtttggc aatgtgctgg tcatcacagc
961 cattgccaag ttcgagcgtc tgcagacggt caccaactac ttcatcactt cactggcctg
1021 tgctgatctg gtcatgggcc tggcagtggt gccctttggg gccgcccata ttcttatgaa
1081 aatgtggact tttggcaact tctggtgcga gttttggact tccattgatg tgctgtgcgt
1141 cacggccagc attgagaccc tgtgcgtgat cgcagtggat cgctactttg ccattacttc
1201 acctttcaag taccagagcc tgctgaccaa gaataaggcc cgggtgatca ttctgatggt
1261 gtggattgtg tcaggcctta cctccttctt gcccattcag atgcactggt accgggccac
1321 ccaccaggaa gccatcaact gctatgccaa tgagacctgc tgtgacttct tcacgaacca
1381 agcctatgcc attgcctctt ccatcgtgtc cttctacgtt cccctggtga tcatggtctt
1441 cgtctactcc agggtctttc aggaggccaa aaggcagctc cagaagattg acaaatctga
1501 gggccgcttc catgtccaga accttagcca ggtggagcag gatgggcgga cggggcatgg
1561 actccgcaga tcttccaagt tctgcttgaa ggagcacaaa gccctcaaga cgttaggcat
1621 catcatgggc actttcaccc tctgctggct gcccttcttc atcgttaaca ttgtgcatgt
1681 gatccaggat aacctcatcc gtaaggaagt ttacatcctc ctaaattgga taggctatgt
1741 caattctggt ttcaatcccc ttatctactg ccggagccca gatttcagga ttgccttcca
1801 ggagcttctg tgcctgcgca ggtcttcttt gaaggcctat gggaatggct actccagcaa
1861 cggcaacaca ggggagcaga gtggatatca cgtggaacag gagaaagaaa ataaactgct
1921 gtgtgaagac ctcccaggca cggaagactt tgtgggccat caaggtactg tgcctagcga
1981 taacattgat tcacaaggga ggaattgtag tacaaatgac tcactgctgg agctggccga
2041 gaagctggcc ggcctggaca tcaacggtgg agctagcggc agccgggcga ccaccctgga
2101 acgtattgaa aagagctttg tgattaccga tccgcgtctg ccggataacc cgattatttt
2161 tgtgagcgat agctttctgc agctgaccga atatagccgt gaagaaattc tgggccgtaa
2221 ctgccgtttt ctgcagggcc cggaaaccga tcgtgcgacc gtgcgtaaaa ttcgtgatgc
2281 gattgataac cagaccgaag tgaccgtgca gctgattaac tataccaaaa gcggcaaaaa
2341 attttggaac ctgtttcatc tgcagccgat gcgtgatcag aaaggcgatg tgcagtattt
2401 tattggcgtg cagctggatg gtaccgaacg tgtgcgtgat gcggcggaac gtgaagctgt
2461 gatgctggtc aagaaaaccg cggaggaaat cgacgaagcc gctaaggaaa atctctactt
2521 ccagatgggc ggaggatccg actacaagga tgacgacgat aagaagctac tgtcttctat
2581 cgaacaagca tgcgatattt gccgacttaa aaagctcaag tgctccaaag aaaaaccgaa
2641 gtgcgccaag tgtctgaaga acaactggga gtgtcgctac tctcccaaaa ccaaaaggtc
2701 tccgctgact agggcacatc tgacagaagt ggaatcaagg ctagaaagac tggaacagct
2761 atttctactg atttttcctc gagaagacct tgacatgatt ttgaaaatgg attctttaca
2821 ggatataaaa gcattgttaa caggattatt tgtacaagat aatgtgaata aagatgccgt
2881 cacagataga ttggcttcag tggagactga tatgcctcta acattgagac agcatagaat
2941 aagtgcgaca tcatcatcgg aagagagtag taacaaaggt caaagacagt tgactgtatc
3001 ggccaatttt aatcaaagtg ggaatattgc tgatagctca ttgtccttca ctttcactaa
3061 cagtagcaac ggtccgaacc tcataacaac tcaaacaaat tctcaagcgc tttcacaacc
3121 aattgcctcc tctaacgttc atgataactt catgaataat gaaatcacgg ctagtaaaat
3181 tgatgatggt aataattcaa aaccactgtc acctggttgg acggaccaaa ctgcgtataa
3241 cgcgtttgga atcactacag ggatgtttaa taccactaca atggatgatg tatataacta
3301 tctattcgat gatgaagata ccccaccaaa cccaaaaaaa gagggcaagc cgatccctaa
3361 ccctctgctg ggcctggaca gcacttaaga attcctcgag ctagagctcg ctgatcagcc
3421 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
3481 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
3541 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
3601 gattgggaag acaatagcag gcatgctggg gaggtaacca cgtgcggacc gagcggccgc
3661 aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg
3721 ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc
3781 gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg catctgtgcg
3841 gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag
3901 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc
3961 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
4021 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
4081 aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
4141 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
4201 actcaactct atctcgggct attcttttga tttataaggg attttgccga tttcggtcta
4261 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
4321 gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
4381 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
4441 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
4501 atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
4561 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
4621 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
4681 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
4741 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
4801 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
4861 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
4921 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
4981 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
5041 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
5101 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
5161 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
5221 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
5281 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
5341 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
5401 tattgctgat aaatctggag ccggtgagcg tggaagccgc ggtatcattg cagcactggg
5461 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
5521 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
5581 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
5641 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
5701 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
5761 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
5821 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
5881 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
5941 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
6001 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
6061 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
6121 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
6181 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
6241 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
6301 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
6361 acggttcctg gccttttgct ggccttttgc tcacatgt
//
