| 质粒名称: | NG-ABE8e |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 138491 |
| 存储实验室: | David Liu |
|
载体骨架基本信息 |
|
| 载体名称: | pCMV with a BR322 origin |
| 载体出品公司: | -- |
| 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: | -- |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | -- |
|
插入基因信息 |
|
| 插入基因名称: | ecTadA(8e)-nSpCas9 (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-ABE8e was a gift from David Liu (Addgene plasmid # 138491 ; http://n2t.net/addgene:138491 ; RRID:Addgene_138491) |
| 参考文献部分: | Phage-assisted evolution of an adenine base editor with improved Cas domain compatibility and activity. Richter MF, Zhao KT, Eton E, Lapinaite A, Newby GA, Thuronyi BW, Wilson C, Koblan LW, Zeng J, Bauer DE, Doudna JA and Liu DR. Nat Biotechnol (2020) 10.1038/s41587-020-0453-z |
买家导航
NGABE8e载体基本信息
NGABE8e载体图谱质粒图谱和多克隆位点信息
NGABE8e载体简介
NGABE8e载体序列
LOCUS NG-ABE8e 8217 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION A-to-G base editor.
ACCESSION .
VERSION .
KEYWORDS NG-ABE8e
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8217)
AUTHORS Richter MF, Zhao KT, Eton E, Lapinaite A, Newby GA, Thuronyi BW,
Wilson C, Koblan LW, Zeng J, Bauer DE, Doudna JA, Liu DR
TITLE Phage-assisted evolution of an adenine base editor with improved Cas
domain compatibility and activity
JOURNAL Nat Biotechnol (2020)
REFERENCE 2 (bases 1 to 8217)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..8217
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 132..335
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 286..306
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 377..396
/label=T7
/note="T7 promoter, forward primer"
promoter 377..395
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 445..465
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 1060..2775
/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 5203..5223
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 5232..5249
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(5272..5289)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 5278..5502
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind complement(5573..5589)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5573..5589)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(5586..5608)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 5597..5613
/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(5621..5651)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5666..5687
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(5804..5821)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5975..6563)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(6055..6074)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(6734..7594)
/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 7357..7376
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(7595..7699)
/gene="bla"
/label=AmpR promoter
primer_bind complement(7778..7797)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 7969..131
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
ORIGIN
1 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
61 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
121 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
181 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
241 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
301 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
361 agagatccgc ggccgctaat acgactcact atagggagag ccgccaccat gaaacggaca
421 gccgacggaa gcgagttcga gtcaccaaag aagaagcgga aagtctctga ggtggagttt
481 tcccacgagt actggatgag acatgccctg accctggcca agagggcacg ggatgagagg
541 gaggtgcctg tgggagccgt gctggtgctg aacaatagag tgatcggcga gggctggaac
601 agagccatcg gcctgcacga cccaacagcc catgccgaaa ttatggccct gagacagggc
661 ggcctggtca tgcagaacta cagactgatt gacgccaccc tgtacgtgac attcgagcct
721 tgcgtgatgt gcgccggcgc catgatccac tctaggatcg gccgcgtggt gtttggcgtg
781 aggaactcaa aaagaggcgc cgcaggctcc ctgatgaacg tgctgaacta ccccggcatg
841 aatcaccgcg tcgaaattac cgagggaatc ctggcagatg aatgtgccgc cctgctgtgc
901 gatttctatc ggatgcctag acaggtgttc aatgctcaga agaaggccca gagctccatc
961 aactccggag gatctagcgg aggctcctct ggctctgaga cacctggcac aagcgagagc
1021 gcaacacctg aaagcagcgg gggcagcagc ggggggtcag acaagaagta cagcatcggc
1081 ctggccatcg gcaccaactc tgtgggctgg gccgtgatca ccgacgagta caaggtgccc
1141 agcaagaaat tcaaggtgct gggcaacacc gaccggcaca gcatcaagaa gaacctgatc
1201 ggagccctgc tgttcgacag cggcgaaaca gccgaggcca cccggctgaa gagaaccgcc
1261 agaagaagat acaccagacg gaagaaccgg atctgctatc tgcaagagat cttcagcaac
1321 gagatggcca aggtggacga cagcttcttc cacagactgg aagagtcctt cctggtggaa
1381 gaggataaga agcacgagcg gcaccccatc ttcggcaaca tcgtggacga ggtggcctac
1441 cacgagaagt accccaccat ctaccacctg agaaagaaac tggtggacag caccgacaag
1501 gccgacctgc ggctgatcta tctggccctg gcccacatga tcaagttccg gggccacttc
1561 ctgatcgagg gcgacctgaa ccccgacaac agcgacgtgg acaagctgtt catccagctg
1621 gtgcagacct acaaccagct gttcgaggaa aaccccatca acgccagcgg cgtggacgcc
1681 aaggccatcc tgtctgccag actgagcaag agcagacggc tggaaaatct gatcgcccag
1741 ctgcccggcg agaagaagaa tggcctgttc ggaaacctga ttgccctgag cctgggcctg
1801 acccccaact tcaagagcaa cttcgacctg gccgaggatg ccaaactgca gctgagcaag
1861 gacacctacg acgacgacct ggacaacctg ctggcccaga tcggcgacca gtacgccgac
1921 ctgtttctgg ccgccaagaa cctgtccgac gccatcctgc tgagcgacat cctgagagtg
1981 aacaccgaga tcaccaaggc ccccctgagc gcctctatga tcaagagata cgacgagcac
2041 caccaggacc tgaccctgct gaaagctctc gtgcggcagc agctgcctga gaagtacaaa
2101 gagattttct tcgaccagag caagaacggc tacgccggct acattgacgg cggagccagc
2161 caggaagagt tctacaagtt catcaagccc atcctggaaa agatggacgg caccgaggaa
2221 ctgctcgtga agctgaacag agaggacctg ctgcggaagc agcggacctt cgacaacggc
2281 agcatccccc accagatcca cctgggagag ctgcacgcca ttctgcggcg gcaggaagat
2341 ttttacccat tcctgaagga caaccgggaa aagatcgaga agatcctgac cttccgcatc
2401 ccctactacg tgggccctct ggccagggga aacagcagat tcgcctggat gaccagaaag
2461 agcgaggaaa ccatcacccc ctggaacttc gaggaagtgg tggacaaggg cgcttccgcc
2521 cagagcttca tcgagcggat gaccaacttc gataagaacc tgcccaacga gaaggtgctg
2581 cccaagcaca gcctgctgta cgagtacttc accgtgtata acgagctgac caaagtgaaa
2641 tacgtgaccg agggaatgag aaagcccgcc ttcctgagcg gcgagcagaa aaaggccatc
2701 gtggacctgc tgttcaagac caaccggaaa gtgaccgtga agcagctgaa agaggactac
2761 ttcaagaaaa tcgagtgctt cgactccgtg gaaatctccg gcgtggaaga tcggttcaac
2821 gcctccctgg gcacatacca cgatctgctg aaaattatca aggacaagga cttcctggac
2881 aatgaggaaa acgaggacat tctggaagat atcgtgctga ccctgacact gtttgaggac
2941 agagagatga tcgaggaacg gctgaaaacc tatgcccacc tgttcgacga caaagtgatg
3001 aagcagctga agcggcggag atacaccggc tggggcaggc tgagccggaa gctgatcaac
3061 ggcatccggg acaagcagtc cggcaagaca atcctggatt tcctgaagtc cgacggcttc
3121 gccaacagaa acttcatgca gctgatccac gacgacagcc tgacctttaa agaggacatc
3181 cagaaagccc aggtgtccgg ccagggcgat agcctgcacg agcacattgc caatctggcc
3241 ggcagccccg ccattaagaa gggcatcctg cagacagtga aggtggtgga cgagctcgtg
3301 aaagtgatgg gccggcacaa gcccgagaac atcgtgatcg aaatggccag agagaaccag
3361 accacccaga agggacagaa gaacagccgc gagagaatga agcggatcga agagggcatc
3421 aaagagctgg gcagccagat cctgaaagaa caccccgtgg aaaacaccca gctgcagaac
3481 gagaagctgt acctgtacta cctgcagaat gggcgggata tgtacgtgga ccaggaactg
3541 gacatcaacc ggctgtccga ctacgatgtg gaccatatcg tgcctcagag ctttctgaag
3601 gacgactcca tcgacaacaa ggtgctgacc agaagcgaca agaaccgggg caagagcgac
3661 aacgtgccct ccgaagaggt cgtgaagaag atgaagaact actggcggca gctgctgaac
3721 gccaagctga ttacccagag aaagttcgac aatctgacca aggccgagag aggcggcctg
3781 agcgaactgg ataaggccgg cttcatcaag agacagctgg tggaaacccg gcagatcaca
3841 aagcacgtgg cacagatcct ggactcccgg atgaacacta agtacgacga gaatgacaag
3901 ctgatccggg aagtgaaagt gatcaccctg aagtccaagc tggtgtccga tttccggaag
3961 gatttccagt tttacaaagt gcgcgagatc aacaactacc accacgccca cgacgcctac
4021 ctgaacgccg tcgtgggaac cgccctgatc aaaaagtacc ctaagctgga aagcgagttc
4081 gtgtacggcg actacaaggt gtacgacgtg cggaagatga tcgccaagag cgagcaggaa
4141 atcggcaagg ctaccgccaa gtacttcttc tacagcaaca tcatgaactt tttcaagacc
4201 gagattaccc tggccaacgg cgagatccgg aagcggcctc tgatcgagac aaacggcgaa
4261 accggggaga tcgtgtggga taagggccgg gattttgcca ccgtgcggaa agtgctgagc
4321 atgccccaag tgaatatcgt gaaaaagacc gaggtgcaga caggcggctt cagcaaagag
4381 tctatcaggc ccaagaggaa cagcgataag ctgatcgcca gaaagaagga ctgggaccct
4441 aagaagtacg gcggcttcgt cagccccacc gtggcctatt ctgtgctggt ggtggccaaa
4501 gtggaaaagg gcaagtccaa gaaactgaag agtgtgaaag agctgctggg gatcaccatc
4561 atggaaagaa gcagcttcga gaagaatccc atcgactttc tggaagccaa gggctacaaa
4621 gaagtgaaaa aggacctgat catcaagctg cctaagtact ccctgttcga gctggaaaac
4681 ggccggaaga gaatgctggc ctctgccaga ttcctgcaga agggaaacga actggccctg
4741 ccctccaaat atgtgaactt cctgtacctg gccagccact atgagaagct gaagggctcc
4801 cccgaggata atgagcagaa acagctgttt gtggaacagc acaagcacta cctggacgag
4861 atcatcgagc agatcagcga gttctccaag agagtgatcc tggccgacgc taatctggac
4921 aaagtgctgt ccgcctacaa caagcaccgg gataagccca tcagagagca ggccgagaat
4981 atcatccacc tgtttaccct gaccaatctg ggagccccta gggccttcaa gtactttgac
5041 accaccatcg accggaaggt gtacaggagc accaaagagg tgctggacgc caccctgatc
5101 caccagagca tcaccggcct gtacgagaca cggatcgacc tgtctcagct gggaggtgac
5161 tctggcggct caaaaagaac cgccgacggc agcgaattcg agcccaagaa gaagaggaaa
5221 gtctaaccgg tcatcatcac catcaccatt gagtttaaac ccgctgatca gcctcgactg
5281 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
5341 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
5401 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
5461 aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa
5521 ccagctgggg ctcgataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
5581 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
5641 taaagtgtaa agcctagggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
5701 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
5761 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
5821 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
5881 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
5941 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
6001 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
6061 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
6121 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
6181 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
6241 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
6301 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
6361 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
6421 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
6481 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
6541 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacactc
6601 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
6661 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
6721 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
6781 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
6841 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
6901 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
6961 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
7021 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
7081 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
7141 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
7201 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
7261 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
7321 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
7381 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
7441 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
7501 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
7561 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
7621 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
7681 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg
7741 gagatcgatc tcccgatccc ctagggtcga ctctcagtac aatctgctct gatgccgcat
7801 agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca
7861 aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttaggg
7921 ttaggcgttt tgcgctgctt cgcgatgtac gggccagata tacgcgttga cattgattat
7981 tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatattgagt
8041 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
8101 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
8161 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatc
//
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