| 质粒名称: | pCMV-dCpf1-BE0 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 107678 |
| 存储实验室: | Jia Chen |
|
载体骨架基本信息 |
|
| 载体名称: | CMV |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | dLbCpf1-BE0,pCMV-APOBEC1-XTEN-dLbCpf1(D832A/E1006A/D1125A)-SV40NLS-SGGS-UGI-SV40NLS |
| 物种来源: | R. norvegicus (rat); Lachnospiraceae bacterium, Bacteriophage PBS2 |
| 插入大小: | 4740 bp |
| 基因突变: | D832A/E925/D1148A |
| GenBank编号 | -- |
| 启动子 | CMV |
| 标签/融合蛋白: | -- |
| 克隆方法: | Gibson克隆 |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | CMV |
| 3' 测序引物 | BGH poly reverse |
|
文献引用方法 |
|
| 材料和方法部分: | pCMV-dCpf1-BE0 was a gift from Jia Chen (Addgene plasmid # 107678 ; http://n2t.net/addgene:107678 ; RRID:Addgene_107678) |
| 参考文献部分: | Base editing with a Cpf1-cytidine deaminase fusion. Li X, Wang Y, Liu Y, Yang B, Wang X, Wei J, Lu Z, Zhang Y, Wu J, Huang X, Yang L, Chen J. Nat Biotechnol. 2018 Mar 19. pii: nbt.4102. doi: 10.1038/nbt.4102. 10.1038/nbt.4102 PubMed 29553573 |
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 pCMV-dCpf1-BE0 8142 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Expresses dLbCpf1-BE0 in mammalian cells.
ACCESSION .
VERSION .
KEYWORDS pCMV-dCpf1-BE0
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8142)
AUTHORS Li X, Wang Y, Liu Y, Yang B, Wang X, Wei J, Lu Z, Zhang Y, Wu J,
Huang X, Yang L, Chen J
TITLE Base editing with a Cpf1-cytidine deaminase fusion.
JOURNAL Nat Biotechnol. 2018 Mar 19. pii: nbt.4102. doi: 10.1038/nbt.4102.
PUBMED 29553573
REFERENCE 2 (bases 1 to 8142)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..8142
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 44..423
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 424..627
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 578..598
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 669..688
/label=T7
/note="T7 promoter, forward primer"
promoter 669..687
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 701..1387
/codon_start=1
/product="cytidine deaminase (C to U editing enzyme) from
rat"
/label=APOBEC-1
/note="can use ssDNA as a substrate (Komor et al., 2016)"
/translation="MSSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWG
GRHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFL
SRYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSN
EAHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHI
LWATGLK"
CDS 5126..5146
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 5159..5407
/codon_start=1
/product="uracil-DNA glycosylase inhibitor from a Bacillus
subtilis bacteriophage (Mol et al., 1995)"
/label=UGI
/translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
CDS 5420..5440
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 5449..5466
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(5489..5506)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 5495..5719
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind complement(5790..5806)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5790..5806)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(5803..5825)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 5814..5830
/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(5838..5868)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5883..5904
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(6021..6038)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(6192..6780)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(6272..6291)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(6951..7811)
/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 7574..7593
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(7812..7916)
/gene="bla"
/label=AmpR promoter
primer_bind complement(7995..8014)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
ORIGIN
1 cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc gttgacattg attattgact
61 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
121 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
181 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
241 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
301 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
361 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
421 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga
481 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
541 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta
601 cggtgggagg tctatataag cagagctggt ttagtgaacc gtcagatccg ctagagatcc
661 gcggccgcta atacgactca ctatagggag agccgccacc atgagctcag agactggccc
721 agtggctgtg gaccccacat tgagacggcg gatcgagccc catgagtttg aggtattctt
781 cgatccgaga gagctccgca aggagacctg cctgctttac gaaattaatt gggggggccg
841 gcactccatt tggcgacata catcacagaa cactaacaag cacgtcgaag tcaacttcat
901 cgagaagttc acgacagaaa gatatttctg tccgaacaca aggtgcagca ttacctggtt
961 tctcagctgg agcccatgcg gcgaatgtag tagggccatc actgaattcc tgtcaaggta
1021 tccccacgtc actctgttta tttacatcgc aaggctgtac caccacgctg acccccgcaa
1081 tcgacaaggc ctgcgggatt tgatctcttc aggtgtgact atccaaatta tgactgagca
1141 ggagtcagga tactgctgga gaaactttgt gaattatagc ccgagtaatg aagcccactg
1201 gcctaggtat ccccatctgt gggtacgact gtacgttctt gaactgtact gcatcatact
1261 gggcctgcct ccttgtctca acattctgag aaggaagcag ccacagctga cattctttac
1321 catcgctctt cagtcttgtc attaccagcg actgccccca cacattctct gggccaccgg
1381 gttgaaaagc ggcagcgaga ctcccgggac ctcagagtcc gccacacccg aaagtatgtc
1441 aaagctggag aaattcacca actgttatag cctgtctaag accctgcgct tcaaggcaat
1501 cccagtgggc aagacacaag agaacattga caacaaacgg ctcctggtgg aggatgagaa
1561 gagggctgaa gattacaagg gcgttaagaa gctgctggat aggtactatc tgtcattcat
1621 caacgatgtc ctccacagta tcaagctgaa gaatctgaac aattacattt ctctgttccg
1681 gaagaagaca cggaccgaga aggagaacaa agagctggag aatctggaga tcaacctgag
1741 gaaagaaata gctaaggctt tcaaagggaa cgagggttac aagtccctgt tcaagaaaga
1801 cattatcgag actattctgc ctgagttcct ggacgataaa gatgagatcg ccctcgtcaa
1861 ttccttcaat gggtttacca cagcctttac cggcttcttc gacaatagag agaatatgtt
1921 ctctgaagag gccaaatcca ctagcatcgc ctttcgctgc ataaacgaga acctgactag
1981 gtacatcagc aatatggaca tctttgagaa agtcgatgcc atattcgaca aacatgaggt
2041 gcaggagatt aaggagaaga tcctgaactc agattacgat gtcgaagatt tcttcgaggg
2101 agagttcttc aacttcgtgc tcacacaaga gggcattgat gtgtacaatg caatcattgg
2161 agggttcgtg acagagagtg gcgagaagat aaagggcctg aacgagtata tcaacctcta
2221 caaccagaaa accaagcaga aactgcctaa gttcaagcca ctgtacaaac aagtgctctc
2281 agatagggaa agcctgagct tctacggtga agggtataca tcagatgaag aagtgctcga
2341 agtgttccgc aacaccctca ataagaacag tgaaatcttc tcttcaatca agaagctgga
2401 gaaactgttc aagaatttcg atgagtactc ctctgccgga atctttgtga agaatggccc
2461 tgcaatatcc actattagca aagacatctt tggcgagtgg aacgttatca gggataagtg
2521 gaatgccgag tacgatgata ttcatctcaa gaagaaagcc gtggttacag agaaatacga
2581 ggatgataga cgcaagagct ttaagaagat tggtagcttc tctctcgaac agctgcagga
2641 gtacgccgac gctgacctgt cagtcgtgga gaaactcaag gagatcataa tccagaaggt
2701 ggatgaaatc tacaaagtgt atggaagctc tgagaaactc ttcgatgcag actttgttct
2761 ggagaagagt ctgaagaaga acgacgcagt ggttgctatc atgaaggacc tgctggattc
2821 tgttaagtct ttcgagaatt acattaaggc attctttggt gaagggaagg agacaaatag
2881 ggacgagagc ttctatggcg actttgttct ggcctacgac atcctcctca aggttgacca
2941 catctatgac gctatacgga attacgttac ccagaagccc tatagcaaag acaagttcaa
3001 gctgtatttc cagaatccac agtttatggg tgggtgggat aaagacaaag aaacagatta
3061 cagggccact atcctgcggt acggcagcaa atactatctg gctatcatgg ataagaagta
3121 cgccaaatgc ctccagaaga tcgacaagga cgacgtgaac ggtaactacg agaagatcaa
3181 ttacaagctc ctgccaggac ctaacaagat gctgcccaag gtgttcttct ccaagaaatg
3241 gatggcctac tataacccaa gcgaggacat tcagaagata tacaagaatg ggacattcaa
3301 gaagggcgat atgttcaacc tcaacgactg ccacaagctg attgatttct tcaaggatag
3361 catttctcgc tatcccaagt ggtctaatgc atacgatttc aacttcagcg agactgagaa
3421 gtacaaagac atcgctggct tctaccggga ggtggaagag caaggctata aggtgtcatt
3481 cgaatccgct tctaagaagg aagtggataa gctcgtggaa gagggtaagc tgtacatgtt
3541 ccagatatac aacaaagact tcagcgataa gagccacggc actccaaacc tccatactat
3601 gtatttcaag ctgctgtttg acgagaacaa ccacggacag attaggctgt caggaggcgc
3661 agaactcttc atgcgcagag cttcactgaa gaaggaggaa ctcgttgtcc acccagccaa
3721 tagccctata gccaataaga atccagacaa tcctaagaaa accactactc tgtcttacga
3781 tgtgtataag gataagagat tctctgaaga tcagtacgaa ctgcacatac ccattgccat
3841 taacaagtgc cctaagaaca tcttcaagat taacacagag gttagagtgc tcctgaaaca
3901 cgacgataac ccttatgtta taggcattgc tcgcggagag agaaacctgc tgtacatcgt
3961 cgtggtggac ggcaaaggca acatcgtgga acagtacagt ctcaatgaaa tcattaacaa
4021 tttcaacgga atccgcatta agaccgacta ccattctctc ctcgacaaga aggagaaaga
4081 aaggttcgaa gcaagacaga attggacaag tatagagaat atcaaagaac tgaaggctgg
4141 gtacatctct caggttgtgc acaagatatg tgagctggtg gagaagtacg acgctgttat
4201 cgccctcgcg gacctgaata gcggcttcaa gaactccagg gtgaaggtgg agaagcaggt
4261 gtatcagaag ttcgagaaga tgctgatcga caagctcaac tatatggtgg acaagaaatc
4321 caatccttgc gctactggtg gagccctgaa gggctatcaa atcaccaata agttcgaatc
4381 tttcaagtct atgagcaccc agaatggctt catcttctac atacccgcat ggctgacatc
4441 caagattgat ccctctaccg gatttgttaa tctgctcaag actaagtaca cctctattgc
4501 tgactcaaag aagttcatat catcatttga ccgcatcatg tacgtgccag aagaggacct
4561 gttcgagttt gccctggatt acaagaattt ctctcggact gacgccgact acatcaagaa
4621 gtggaagctc tactcttatg gtaatcggat tcgcatattc cgcaatccca agaagaataa
4681 cgtgttcgat tgggaggaag tttgcctcac cagcgcttac aaggagctgt tcaataagta
4741 tgggattaac taccagcagg gcgacataag agccctgctg tgcgaacaat ctgataaggc
4801 attctattcc tctttcatgg cactgatgtc actgatgctg caaatgcgca attccatcac
4861 cggaagaaca gacgtggcct ttctgatctc tcctgtcaag aactcagatg gcatcttcta
4921 cgattcccgc aactatgaag cacaggagaa tgctatcctg cctaagaatg ccgatgcaaa
4981 tggagcctat aacatcgcca gaaaggtcct ctgggccata ggacaattca agaaagctga
5041 agatgagaag ctggacaagg tgaagatcgc catttcaaac aaagagtggc tcgaatatgc
5101 tcagacctca gtgaagcatg gatcacccaa gaagaaacgg aaagtgtctg gtggttctac
5161 taatctgtca gatattattg aaaaggagac cggtaagcaa ctggttatcc aggaatccat
5221 cctcatgctc ccagaggagg tggaagaagt cattgggaac aagccggaaa gcgatatact
5281 cgtgcacacc gcctacgacg agagcaccga cgagaatgtc atgcttctga ctagcgacgc
5341 ccctgaatac aagccttggg ctctggtcat acaggatagc aacggtgaga acaagattaa
5401 gatgctctct ggtggttctc ccaagaagaa gaggaaagtc taaccggtca tcatcaccat
5461 caccattgag tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
5521 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
5581 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
5641 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
5701 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc gataccgtcg
5761 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
5821 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctagggtgcc
5881 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
5941 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
6001 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
6061 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
6121 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
6181 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
6241 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
6301 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
6361 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
6421 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
6481 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
6541 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
6601 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
6661 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
6721 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
6781 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
6841 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
6901 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
6961 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
7021 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
7081 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
7141 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
7201 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
7261 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
7321 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
7381 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
7441 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
7501 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
7561 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
7621 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
7681 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
7741 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
7801 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
7861 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
7921 tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atcgatctcc cgatccccta
7981 gggtcgactc tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct
8041 gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa
8101 ggcttgaccg acaattgcat gaagaatctg cttagggtta gg
//
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