| 质粒名称: | pCAGGS-6011nes |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 108652 |
| 存储实验室: | Michiyuki Matsuda |
|
载体骨架基本信息 |
|
| 载体名称: | pCAGGS |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 4739 bp |
| 完整质粒大小: | 7987 bp |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | -- |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | hyBRET-ERK-EV |
| 物种来源: | S. cerevisiae (budding yeast), Synthetic; aequorea coerulescens |
| 插入大小: | 3200 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | pCAGGS |
| 标签/融合蛋白: | XFP（插入件上的N端子） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | EcoRI(未破坏) |
| 3' 克隆位点: | SalI (未破坏) |
| 5' 测序引物 | pCAGGSseq_F |
| 3' 测序引物 | pCAGGSseq_R |
|
文献引用方法 |
|
| 材料和方法部分: | pCAGGS-6011nes was a gift from Michiyuki Matsuda (Addgene plasmid # 108652 ; http://n2t.net/addgene:108652 ; RRID:Addgene_108652) |
| 参考文献部分: | A platform of BRET-FRET hybrid biosensors for optogenetics, chemical screening, and in vivo imaging. Komatsu N, Terai K, Imanishi A, Kamioka Y, Sumiyama K, Jin T, Okada Y, Nagai T, Matsuda M. Sci Rep. 2018 Jun 12;8(1):8984. doi: 10.1038/s41598-018-27174-x. 10.1038/s41598-018-27174-x PubMed 29895862 |
买家导航
pCAGGS6011nes载体基本信息
pCAGGS6011nes载体图谱质粒图谱和多克隆位点信息
pCAGGS6011nes载体简介
pCAGGS6011nes载体序列
LOCUS pCAGGS-6011nes 7698 bp ds-DNA circular SYN 05-FEB-2020
DEFINITION An ERK biosensor based on fluorescence resonance energy transfer
(FRET) and those based on bioluminescence energy transfer (BRET).
ACCESSION .
VERSION .
KEYWORDS pCAGGS-6011nes
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7698)
AUTHORS Komatsu N, Terai K, Imanishi A, Kamioka Y, Sumiyama K, Jin T, Okada
Y, Nagai T, Matsuda M
TITLE A platform of BRET-FRET hybrid biosensors for optogenetics, chemical
screening, and in vivo imaging.
JOURNAL Sci Rep. 2018 Jun 12;8(1):8984. doi: 10.1038/s41598-018-27174-x.
PUBMED 29895862
REFERENCE 2 (bases 1 to 7698)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Feb 5, 2020 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7698
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 9..388
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 390..665
/label=chicken beta-actin promoter
primer_bind 1606..1628
/label=pCAGGS-5
/note="Chimeric intron in CAG promoter, forward primer"
primer_bind 1694..1713
/label=pCAG-F
/note="Rabbit beta-globin intron, for pCAG plasmids,
forward primer"
CDS 1744..2262
/codon_start=1
/product="N-terminal fragment of mVenus for use in
bimolecular fluorescence complementation (BiFC) (Kodama and
Hu, 2010)"
/label=VN173
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KLLCTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
ANFKIRHNIE"
CDS 4678..4704
/codon_start=1
/product="nuclear export signal from the HIV Rev protein
(Fischer et al., 1995)"
/label=NES
/translation="LPPLERLTL"
primer_bind complement(4720..4739)
/label=Bglob-pA-R
/note="Rabbit beta-globin polyA region, reverse primer"
polyA_site 4785..4840
/label=-beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal"
primer_bind complement(4839..4858)
/label=rbglobpA-R
/note="Rabbit beta-globin polyA, reverse primer. Also
called rb-glob-pA-term-R"
primer_bind complement(5201..5217)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5201..5217)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(5214..5236)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 5225..5241
/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(5249..5279)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5294..5315
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 5374..5569
/label=SV40 promoter
/note="SV40 early promoter"
rep_origin 5420..5555
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 5482..5501
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
polyA_signal 5575..5709
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(5612..5631)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 5666..5685
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(5777..5794)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5948..6536)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(6028..6047)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(6707..7567)
/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 7330..7349
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(7568..7672)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 ggtcgatcga cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt
61 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
121 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
181 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
241 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
301 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
361 ctacgtatta gtcatcgcta ttaccatggt cgaggtgagc cccacgttct gcttcactct
421 ccccatctcc cccccctccc cacccccaat tttgtattta tttatttttt aattattttg
481 tgcagcgatg ggggcggggg gggggggggg gcgcgccagg cggggcgggg cggggcgagg
541 ggcggggcgg ggcgaggcgg agaggtgcgg cggcagccaa tcagagcggc gcgctccgaa
601 agtttccttt tatggcgagg cggcggcggc ggcggcccta taaaaagcga agcgcgcggc
661 gggcgggagt cgctgcgcgc tgccttcgcc ccgtgccccg ctccgccgcc gcctcgcgcc
721 gcccgccccg gctctgactg accgcgttac tcccacaggt gagcgggcgg gacggccctt
781 ctcctccggg ctgtaattag cgcttggttt aatgacggct tgtttctttt ctgtggctgc
841 gtgaaagcct tgaggggctc cgggagggcc ctttgtgcgg ggggagcggc tcggggggtg
901 cgtgcgtgtg tgtgtgcgtg gggagcgccg cgtgcggctc cgcgctgccc ggcggctgtg
961 agcgctgcgg gcgcggcgcg gggctttgtg cgctccgcag tgtgcgcgag gggagcgcgg
1021 ccgggggcgg tgccccgcgg tgcggggggg gctgcgaggg gaacaaaggc tgcgtgcggg
1081 gtgtgtgcgt gggggggtga gcagggggtg tgggcgcgtc ggtcgggctg caaccccccc
1141 tgcacccccc tccccgagtt gctgagcacg gcccggcttc gggtgcgggg ctccgtacgg
1201 ggcgtggcgc ggggctcgcc gtgccgggcg gggggtggcg gcaggtgggg gtgccgggcg
1261 gggcggggcc gcctcgggcc ggggagggct cgggggaggg gcgcggcggc ccccggagcg
1321 ccggcggctg tcgaggcgcg gcgagccgca gccattgcct tttatggtaa tcgtgcgaga
1381 gggcgcaggg acttcctttg tcccaaatct gtgcggagcc gaaatctggg aggcgccgcc
1441 gcaccccctc tagcgggcgc ggggcgaagc ggtgcggcgc cggcaggaag gaaatgggcg
1501 gggagggcct tcgtgcgtcg ccgcgccgcc gtccccttct ccctctccag cctcggggct
1561 gtccgcgggg ggacggctgc cttcgggggg gacggggcag ggcggggttc ggcttctggc
1621 gtgtgaccgg cggctctagc tagagcctct gctaaccatg ttcatgcctt cttctttttc
1681 ctacagctcc tgggcaacgt gctggttatt gtgctgtctc atcattttgg caaagaattc
1741 ggcatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
1801 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
1861 tacggcaagc tgaccctgaa gcttctatgc accaccggca agctgcccgt gccctggccc
1921 accctcgtga ccaccctggg ctacggcctg cagtgcttcg cccgctaccc cgaccacatg
1981 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
2041 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
2101 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
2161 cacaagctgg agtacaacta caacagccac aacgtctata tcaccgccga caagcagaag
2221 aacggcatca aggccaactt caagatccgc cacaacatcg aggacggcgg cgtgcagctc
2281 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
2341 cactacctga gctaccagtc cgccctgttc aaagacccca acgagaagcg cgatcacatg
2401 gtcctgctgg agttcctgac cgccgccggg atcactgagg gcatgaacga gctgtacctc
2461 gagatggcgg acgaggagaa gctgccgccc ggctgggaga agcgcatgag ccgcagctca
2521 ggccgagtgt actacttcaa ccacatcact aacgccagcc agtgggagcg gcccagcggc
2581 aacagcagca gtggtggcaa aaacgggcag ggggagcctg ccaggggtac cagtgctggt
2641 ggtagtgctg gtggtagtgc tggtggtagt gctggtggta gtgctggtgg ttccggcagt
2701 gctggtggta gtgctggtgg tagtaccagt gctggtggta gtgctggtgg tagtgctggt
2761 ggtagtgctg gtggtagtgc tggtggttcc ggcagtgctg gtggtagtgc tggtggtagt
2821 accagtgctg gtggtagtgc tggtggtagt gctggtggta gtgctggtgg tagtgctggt
2881 ggttccggca gtgctggtgg tagtgctggt ggtagtacca gtgctggtgg tagtgctggt
2941 ggtagtgctg gtggtagtgc tggtggtagt gctggtggtt ccggaccaga tgtccctaga
3001 actccagtgg ataaagcaaa gctgtcattc caatttccgg gcggccgcat ggtgagcaag
3061 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtcaac
3121 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
3181 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
3241 ctgtcctggg gcgtgcagtg cttcgcccgc taccccgacc acatgaagca gcacgacttc
3301 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
3361 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
3421 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
3481 aactacttca gcgggaacgt ctatatcacc gccgacaagc agaagaacgg catcaaggcc
3541 aacttcaaga tccgccacaa catcgaggac ggcggcgtgc agctcgccga ccactaccag
3601 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc
3661 cagtccgcct taagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
3721 ttgaccgccg ccgggggtac caaggtgtac gaccccgagc agaggaagag gatgatcacc
3781 ggcccccagt ggtgggccag gtgcaagcag atgaacgtgc tggacagctt catcaactac
3841 tacgacagcg agaagcacgc cgagaacgcc gtgatcttcc tgcacggcaa cgccactagc
3901 agctacctgt ggaggcacgt ggtgccccac atcgagcccg tggccaggtg catcatcccc
3961 gatctgatcg gcatgggcaa gagcggcaag agcggcaacg gcagctacag gctgctggac
4021 cactacaagt acctgaccgc ctggttcgag cttctgaacc tgcccaagaa gatcatcttc
4081 gtgggccacg actggggcgc cgccctggcc ttccactacg cctacgagca ccaggacagg
4141 atcaaggcca tcgtgcacat ggagagcgtg gtggacgtga tcgagagctg ggacgagtgg
4201 ccagacatcg aggaggacat cgccctgatc aagagcgagg agggcgagaa gatggtgctg
4261 gagaacaact tcttcgtgga gaccgtgctg cccagcaaga tcatgagaaa gctggagccc
4321 gaggagttcg ccgcctacct ggagcccttc aaggagaagg gcgaggtgag aagacccacc
4381 ctgagctggc ccagagagat ccccctggtg aagggcggca agcccgacgt ggtgcagatc
4441 gtgagaaact acaacgccta cctgagagcc agcgacgacc tgcccaagct gttcatcgag
4501 ggcgaccccg gcttcttcag caacgccatc gtggagggcg ccaagaagtt ccccaacacc
4561 gagttcgtga aggtgaaggg cctgcacttc ctccaggagg acgcccccga cgagatgggc
4621 aagtacatca agagcttcgt ggagagagtg ctgaagaacg agcagtctag actgcagctg
4681 cctcctctgg aacgcctgac tctggattaa gtcgacggat ctttttccct ctgccaaaaa
4741 ttatggggac atcatgaagc cccttgagca tctgacttct ggctaataaa ggaaatttat
4801 tttcattgca atagtgtgtt ggaatttttt gtgtctctca ctcggaagga catatgggag
4861 ggcaaatcat ttaaaacatc agaatgagta tttggtttag agtttggcaa catatgccca
4921 tatgctggct gccatgaaca aaggttggct ataaagaggt catcagtata tgaaacagcc
4981 ccctgctgtc cattccttat tccatagaaa agccttgact tgaggttaga ttttttttat
5041 attttgtttt gtgttatttt tttctttaac atccctaaaa ttttccttac atgttttact
5101 agccagattt ttcctcctct cctgactact cccagtcata gctgtccctc ttctcttatg
5161 gagatccctc gacctgcagc ccaagcttgg cgtaatcatg gtcatagctg tttcctgtgt
5221 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
5281 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
5341 tccagtcggg aaacctgtcg tgccagcgga tccgcatctc aattagtcag caaccatagt
5401 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
5461 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
5521 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctaacttg
5581 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
5641 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
5701 gtctggatcc gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
5761 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
5821 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
5881 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
5941 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
6001 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
6061 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
6121 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
6181 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
6241 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
6301 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
6361 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
6421 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
6481 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
6541 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
6601 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
6661 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
6721 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
6781 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
6841 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
6901 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
6961 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
7021 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
7081 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
7141 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
7201 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
7261 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
7321 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
7381 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
7441 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
7501 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
7561 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
7621 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
7681 cccgaaaagt gccacctg
//
