| 质粒名称: | NG-ABEmax |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 124163 |
| 存储实验室: | David Liu |
|
载体骨架基本信息 |
|
| 载体名称: | pCMV |
| 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: | -- |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | TadA-TadA(evo)-Cas9-NG |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | -- |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | -- |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | NG-ABEmax was a gift from David Liu (Addgene plasmid # 124163 ; http://n2t.net/addgene:124163 ; RRID:Addgene_124163) |
| 参考文献部分: | Circularly permuted and PAM-modified Cas9 variants broaden the targeting scope of base editors. Huang TP, Zhao KT, Miller SM, Gaudelli NM, Oakes BL, Fellmann C, Savage DF, Liu DR. Nat Biotechnol. 2019 May 20. pii: 10.1038/s41587-019-0134-y. doi: 10.1038/s41587-019-0134-y. 10.1038/s41587-019-0134-y PubMed 31110355 |
买家导航
NGABEmax载体基本信息
NGABEmax载体图谱质粒图谱和多克隆位点信息
NGABEmax载体简介
NGABEmax载体序列
LOCUS NG-ABEmax 8811 bp ds-DNA circular SYN 09-APR-2019
DEFINITION A-to-G base editor.
ACCESSION .
VERSION .
KEYWORDS NG-ABEmax
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8811)
TITLE Liu lab base editing plasmids
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 8811)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 9, 2019 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..8811
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 155..534
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 535..738
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 689..709
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 780..799
/label=T7
/note="T7 promoter, forward primer"
promoter 780..798
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 848..868
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 2057..3772
/codon_start=1
/product="N-terminal portion of Streptococcus pyogenes Cas9
(Zetsche et al., 2015)"
/label=Cas9(N)
/translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIE"
CDS 6200..6220
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 6229..6246
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(6269..6286)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 6275..6499
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind complement(6570..6586)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(6570..6586)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(6583..6605)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 6594..6610
/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(6618..6648)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 6663..6684
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(6801..6818)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(6972..7560)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(7052..7071)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(7731..8591)
/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 8354..8373
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(8592..8696)
/gene="bla"
/label=AmpR promoter
primer_bind complement(8775..8794)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
ORIGIN
1 ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca
61 acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagggttag gcgttttgcg
121 ctgcttcgcg atgtacgggc cagatatacg cgttgacatt gattattgac tagttattaa
181 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
241 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
301 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
361 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
421 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
481 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg
541 cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt
601 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca
661 aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag
721 gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagagatc cgcggccgct
781 aatacgactc actataggga gagccgccac catgaaacgg acagccgacg gaagcgagtt
841 cgagtcacca aagaagaagc ggaaagtctc tgaagtcgag tttagccacg agtattggat
901 gaggcacgca ctgaccctgg caaagcgagc atgggatgaa agagaagtcc ccgtgggcgc
961 cgtgctggtg cacaacaata gagtgatcgg agagggatgg aacaggccaa tcggccgcca
1021 cgaccctacc gcacacgcag agatcatggc actgaggcag ggaggcctgg tcatgcagaa
1081 ttaccgcctg atcgatgcca ccctgtatgt gacactggag ccatgcgtga tgtgcgcagg
1141 agcaatgatc cacagcagga tcggaagagt ggtgttcgga gcacgggacg ccaagaccgg
1201 cgcagcaggc tccctgatgg atgtgctgca ccaccccggc atgaaccacc gggtggagat
1261 cacagaggga atcctggcag acgagtgcgc cgccctgctg agcgatttct ttagaatgcg
1321 gagacaggag atcaaggccc agaagaaggc acagagctcc accgactctg gaggatctag
1381 cggaggatcc tctggaagcg agacaccagg cacaagcgag tccgccacac cagagagctc
1441 cggcggctcc tccggaggat cctctgaggt ggagttttcc cacgagtact ggatgagaca
1501 tgccctgacc ctggccaaga gggcacgcga tgagagggag gtgcctgtgg gagccgtgct
1561 ggtgctgaac aatagagtga tcggcgaggg ctggaacaga gccatcggcc tgcacgaccc
1621 aacagcccat gccgaaatta tggccctgag acagggcggc ctggtcatgc agaactacag
1681 actgattgac gccaccctgt acgtgacatt cgagccttgc gtgatgtgcg ccggcgccat
1741 gatccactct aggatcggcc gcgtggtgtt tggcgtgagg aacgcaaaaa ccggcgccgc
1801 aggctccctg atggacgtgc tgcactaccc cggcatgaat caccgcgtcg aaattaccga
1861 gggaatcctg gcagatgaat gtgccgccct gctgtgctat ttctttcgga tgcctagaca
1921 ggtgttcaat gctcagaaga aggcccagag ctccaccgac tccggaggat ctagcggagg
1981 ctcctctggc tctgagacac ctggcacaag cgagagcgca acacctgaaa gcagcggggg
2041 cagcagcggg gggtcagaca agaagtacag catcggcctg gccatcggca ccaactctgt
2101 gggctgggcc gtgatcaccg acgagtacaa ggtgcccagc aagaaattca aggtgctggg
2161 caacaccgac cggcacagca tcaagaagaa cctgatcgga gccctgctgt tcgacagcgg
2221 cgaaacagcc gaggccaccc ggctgaagag aaccgccaga agaagataca ccagacggaa
2281 gaaccggatc tgctatctgc aagagatctt cagcaacgag atggccaagg tggacgacag
2341 cttcttccac agactggaag agtccttcct ggtggaagag gataagaagc acgagcggca
2401 ccccatcttc ggcaacatcg tggacgaggt ggcctaccac gagaagtacc ccaccatcta
2461 ccacctgaga aagaaactgg tggacagcac cgacaaggcc gacctgcggc tgatctatct
2521 ggccctggcc cacatgatca agttccgggg ccacttcctg atcgagggcg acctgaaccc
2581 cgacaacagc gacgtggaca agctgttcat ccagctggtg cagacctaca accagctgtt
2641 cgaggaaaac cccatcaacg ccagcggcgt ggacgccaag gccatcctgt ctgccagact
2701 gagcaagagc agacggctgg aaaatctgat cgcccagctg cccggcgaga agaagaatgg
2761 cctgttcgga aacctgattg ccctgagcct gggcctgacc cccaacttca agagcaactt
2821 cgacctggcc gaggatgcca aactgcagct gagcaaggac acctacgacg acgacctgga
2881 caacctgctg gcccagatcg gcgaccagta cgccgacctg tttctggccg ccaagaacct
2941 gtccgacgcc atcctgctga gcgacatcct gagagtgaac accgagatca ccaaggcccc
3001 cctgagcgcc tctatgatca agagatacga cgagcaccac caggacctga ccctgctgaa
3061 agctctcgtg cggcagcagc tgcctgagaa gtacaaagag attttcttcg accagagcaa
3121 gaacggctac gccggctaca ttgacggcgg agccagccag gaagagttct acaagttcat
3181 caagcccatc ctggaaaaga tggacggcac cgaggaactg ctcgtgaagc tgaacagaga
3241 ggacctgctg cggaagcagc ggaccttcga caacggcagc atcccccacc agatccacct
3301 gggagagctg cacgccattc tgcggcggca ggaagatttt tacccattcc tgaaggacaa
3361 ccgggaaaag atcgagaaga tcctgacctt ccgcatcccc tactacgtgg gccctctggc
3421 caggggaaac agcagattcg cctggatgac cagaaagagc gaggaaacca tcaccccctg
3481 gaacttcgag gaagtggtgg acaagggcgc ttccgcccag agcttcatcg agcggatgac
3541 caacttcgat aagaacctgc ccaacgagaa ggtgctgccc aagcacagcc tgctgtacga
3601 gtacttcacc gtgtataacg agctgaccaa agtgaaatac gtgaccgagg gaatgagaaa
3661 gcccgccttc ctgagcggcg agcagaaaaa ggccatcgtg gacctgctgt tcaagaccaa
3721 ccggaaagtg accgtgaagc agctgaaaga ggactacttc aagaaaatcg agtgcttcga
3781 ctccgtggaa atctccggcg tggaagatcg gttcaacgcc tccctgggca cataccacga
3841 tctgctgaaa attatcaagg acaaggactt cctggacaat gaggaaaacg aggacattct
3901 ggaagatatc gtgctgaccc tgacactgtt tgaggacaga gagatgatcg aggaacggct
3961 gaaaacctat gcccacctgt tcgacgacaa agtgatgaag cagctgaagc ggcggagata
4021 caccggctgg ggcaggctga gccggaagct gatcaacggc atccgggaca agcagtccgg
4081 caagacaatc ctggatttcc tgaagtccga cggcttcgcc aacagaaact tcatgcagct
4141 gatccacgac gacagcctga cctttaaaga ggacatccag aaagcccagg tgtccggcca
4201 gggcgatagc ctgcacgagc acattgccaa tctggccggc agccccgcca ttaagaaggg
4261 catcctgcag acagtgaagg tggtggacga gctcgtgaaa gtgatgggcc ggcacaagcc
4321 cgagaacatc gtgatcgaaa tggccagaga gaaccagacc acccagaagg gacagaagaa
4381 cagccgcgag agaatgaagc ggatcgaaga gggcatcaaa gagctgggca gccagatcct
4441 gaaagaacac cccgtggaaa acacccagct gcagaacgag aagctgtacc tgtactacct
4501 gcagaatggg cgggatatgt acgtggacca ggaactggac atcaaccggc tgtccgacta
4561 cgatgtggac catatcgtgc ctcagagctt tctgaaggac gactccatcg acaacaaggt
4621 gctgaccaga agcgacaaga accggggcaa gagcgacaac gtgccctccg aagaggtcgt
4681 gaagaagatg aagaactact ggcggcagct gctgaacgcc aagctgatta cccagagaaa
4741 gttcgacaat ctgaccaagg ccgagagagg cggcctgagc gaactggata aggccggctt
4801 catcaagaga cagctggtgg aaacccggca gatcacaaag cacgtggcac agatcctgga
4861 ctcccggatg aacactaagt acgacgagaa tgacaagctg atccgggaag tgaaagtgat
4921 caccctgaag tccaagctgg tgtccgattt ccggaaggat ttccagtttt acaaagtgcg
4981 cgagatcaac aactaccacc acgcccacga cgcctacctg aacgccgtcg tgggaaccgc
5041 cctgatcaaa aagtacccta agctggaaag cgagttcgtg tacggcgact acaaggtgta
5101 cgacgtgcgg aagatgatcg ccaagagcga gcaggaaatc ggcaaggcta ccgccaagta
5161 cttcttctac agcaacatca tgaacttttt caagaccgag attaccctgg ccaacggcga
5221 gatccggaag cggcctctga tcgagacaaa cggcgaaacc ggggagatcg tgtgggataa
5281 gggccgggat tttgccaccg tgcggaaagt gctgagcatg ccccaagtga atatcgtgaa
5341 aaagaccgag gtgcagacag gcggcttcag caaagagtct atcaggccca agaggaacag
5401 cgataagctg atcgccagaa agaaggactg ggaccctaag aagtacggcg gcttcgtcag
5461 ccccaccgtg gcctattctg tgctggtggt ggccaaagtg gaaaagggca agtccaagaa
5521 actgaagagt gtgaaagagc tgctggggat caccatcatg gaaagaagca gcttcgagaa
5581 gaatcccatc gactttctgg aagccaaggg ctacaaagaa gtgaaaaagg acctgatcat
5641 caagctgcct aagtactccc tgttcgagct ggaaaacggc cggaagagaa tgctggcctc
5701 tgccagattc ctgcagaagg gaaacgaact ggccctgccc tccaaatatg tgaacttcct
5761 gtacctggcc agccactatg agaagctgaa gggctccccc gaggataatg agcagaaaca
5821 gctgtttgtg gaacagcaca agcactacct ggacgagatc atcgagcaga tcagcgagtt
5881 ctccaagaga gtgatcctgg ccgacgctaa tctggacaaa gtgctgtccg cctacaacaa
5941 gcaccgggat aagcccatca gagagcaggc cgagaatatc atccacctgt ttaccctgac
6001 caatctggga gcccctaggg ccttcaagta ctttgacacc accatcgacc ggaaggtgta
6061 caggagcacc aaagaggtgc tggacgccac cctgatccac cagagcatca ccggcctgta
6121 cgagacacgg atcgacctgt ctcagctggg aggtgactct ggcggctcaa aaagaaccgc
6181 cgacggcagc gaattcgagc ccaagaagaa gaggaaagtc taaccggtca tcatcaccat
6241 caccattgag tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
6301 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
6361 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
6421 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
6481 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc gataccgtcg
6541 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
6601 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctagggtgcc
6661 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
6721 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
6781 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
6841 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
6901 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
6961 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
7021 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
7081 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
7141 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
7201 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
7261 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
7321 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
7381 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
7441 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
7501 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
7561 gaagatcctt tgatcttttc tacggggtct gacactcagt ggaacgaaaa ctcacgttaa
7621 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
7681 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
7741 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
7801 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
7861 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
7921 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
7981 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
8041 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
8101 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
8161 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
8221 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
8281 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
8341 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
8401 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
8461 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
8521 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
8581 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
8641 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
8701 tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atcgatctcc cgatccccta
8761 gggtcgactc tcagtacaat ctgctctgat gccgcatagt taagccagta t
//
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