| 质粒名称: | CMV-CASANOVA |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 113035 |
| 存储实验室: | Dominik Niopek |
|
载体骨架基本信息 |
|
| 载体名称: | pcDNA3.1 |
| 空载体大小: | 6136bp |
| 完整质粒大小: | 6139 bp |
| 载体类型: | 哺乳细胞表达、CRISPR, Synthetic Biology |
| 载体抗性: | 氨苄霉素 |
| 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: | CASANOVA / engineered AcrIIA4-LOV2 hybrid for optogenetic control of CRISPR/SpyCas9 /inhibits SpyCas9 in the absence of light and releases Cas9 activity upon photoactivation |
|
文献引用方法 |
|
| 材料和方法部分: | CMV-CASANOVA was a gift from Dominik Niopek (Addgene plasmid # 113035 ; http://n2t.net/addgene:113035 ; RRID:Addgene_113035) |
| 参考文献部分: | Engineered anti-CRISPR proteins for optogenetic control of CRISPR-Cas9. Bubeck F, Hoffmann MD, Harteveld Z, Aschenbrenner S, Bietz A, Waldhauer MC, Borner K, Fakhiri J, Schmelas C, Dietz L, Grimm D, Correia BE, Eils R, Niopek D. Nat Methods. 2018 Oct 30. pii: 10.1038/s41592-018-0178-9. doi: 10.1038/s41592-018-0178-9. 10.1038/s41592-018-0178-9 PubMed 30377362 |
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 CMV-CASANOVA 6139 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION CASANOVA (AcrIIA4-LOV2 hybrid) expression in mammalian cells;
enables optogenetic control of SpyCas9.
ACCESSION .
VERSION .
KEYWORDS CMV-CASANOVA
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6139)
AUTHORS Bubeck F, Hoffmann MD, Harteveld Z, Aschenbrenner S, Bietz A,
Waldhauer MC, Borner K, Fakhiri J, Schmelas C, Dietz L, Grimm D,
Correia BE, Eils R, Niopek D
TITLE Engineered anti-CRISPR proteins for optogenetic control of
CRISPR-Cas9.
JOURNAL Nat Methods. 2018 Oct 30. pii: 10.1038/s41592-018-0178-9. doi:
10.1038/s41592-018-0178-9.
PUBMED 30377362
REFERENCE 2 (bases 1 to 6139)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..6139
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(33..52)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 224..603
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 604..807
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 758..778
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 852..871
/label=T7
/note="T7 promoter, forward primer"
promoter 852..870
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 896..916
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
primer_bind complement(1719..1736)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 1725..1949
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 1995..2423
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2082..2101)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2292..2313
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(2432..2452)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 2437..2766
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 2617..2752
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2679..2698
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 2833..3627
/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
VLFVKTDLSGALNELQDEAARLS*LATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(2887..2906)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3497..3516
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 3801..3922
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3838..3857)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3892..3911
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(3971..3987)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3971..3987)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(3984..4006)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 3995..4011
/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(4019..4049)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4064..4085
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4202..4219)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4373..4961)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4453..4472)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5132..5992)
/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 5755..5774
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5993..6097)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 agatctcccg atcccctatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
61 agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt
121 taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct tagggttagg
181 cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc gttgacattg attattgact
241 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
301 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
361 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
421 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
481 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
541 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
601 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga
661 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
721 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta
781 cggtgggagg tctatataag cagagctctc tggctaacta gagaacccac tgcttactgg
841 cttatcgaaa ttaatacgac tcactatagg gagacccaag ctggctagca tggccccgaa
901 gaaaaagcga aaggtgggcg gaagcggagg ttccatgaac attaacgacc tgattcggga
961 aataaagaac aaggactaca cggtcaagct gtcaggcacc gactccaact ctataacaca
1021 gctcattata agggttaata atgatggcaa cgaatatgtc atctctgaat ccgagaatga
1081 atcaatcgtg gagaaattca tcagtgcatt taaaaacggc tggcttgcta caactttgga
1141 gcggattgag aagaatttcg tcataactga tcccaggttg cctgataatc ctataatatt
1201 cgcctccgat agttttcttc aacttacgga gtactccaga gaggagatac tgggaaggaa
1261 ttgcagattc ctccaaggtc cagaaactga tagagctact gtgaggaaga tacgggatgc
1321 cattgacaat caaactgaag tgacagtgca acttatcaat tacaccaagt cagggaaaaa
1381 attctggaac ctgttccatt tgcaacccat gagggaccaa aagggcgatg tgcaatattt
1441 cataggcgtt caacttgatg gtactgaaca cgtcagagat gcggcggagc gagaaggagt
1501 gatgctgatt aagaaaacag cagagaatat cgacgaagcg gcgaaagagt tgtacgaaga
1561 tgaggaagag ttctacaatg atatgcaaac tatcacactg aaatctgagc tgaactagct
1621 cgagcggccg ccactgtgct ggatatctgc agaattccac cacactggac tagtggatcc
1681 gagctcggta ccaagcttaa gtttaaaccg ctgatcagcc tcgactgtgc cttctagttg
1741 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
1801 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
1861 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
1921 gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc
1981 tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
2041 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
2101 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
2161 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
2221 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
2281 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
2341 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
2401 ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag
2461 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
2521 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
2581 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
2641 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
2701 gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
2761 tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga
2821 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
2881 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
2941 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
3001 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtgac tggccacgac gggcgttcct
3061 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
3121 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
3181 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
3241 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
3301 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
3361 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
3421 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
3481 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
3541 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
3601 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
3661 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
3721 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
3781 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca
3841 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
3901 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg
3961 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
4021 acatgcgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
4081 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
4141 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
4201 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
4261 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
4321 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
4381 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
4441 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
4501 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
4561 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
4621 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
4681 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
4741 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
4801 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
4861 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
4921 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
4981 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
5041 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
5101 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
5161 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
5221 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
5281 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
5341 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
5401 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
5461 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
5521 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
5581 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
5641 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
5701 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
5761 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
5821 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
5881 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
5941 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
6001 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
6061 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
6121 ctgacgtcga cggatcggg
//
