| 质粒名称: | VRQR-ABEmax |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 119811 |
| 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: | -- |
| 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: | ecTadA-ecTadA*(7.10)-nSpCas9 (VRQR) |
| 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: | VRQR-ABEmax was a gift from David Liu (Addgene plasmid # 119811 ; http://n2t.net/addgene:119811 ; RRID:Addgene_119811) |
| 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
VRQRABEmax载体基本信息
VRQRABEmax载体图谱质粒图谱和多克隆位点信息
VRQRABEmax载体简介
VRQRABEmax载体序列
LOCUS VRQR-ABEmax 8811 bp ds-DNA circular SYN 20-DEC-2018
DEFINITION A-to-G base editor.
ACCESSION .
VERSION .
KEYWORDS VRQR-ABEmax
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8811)
TITLE Liu lab base editing plasmids
REFERENCE 2 (bases 1 to 8811)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Dec 20, 2018 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 140..519
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 520..723
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 674..694
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 765..784
/label=T7
/note="T7 promoter, forward primer"
promoter 765..783
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 833..853
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 2042..3757
/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 6185..6205
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 6214..6231
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(6254..6271)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 6260..6484
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind complement(6555..6571)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(6555..6571)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(6568..6590)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 6579..6595
/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(6603..6633)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 6648..6669
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(6786..6803)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(6957..7545)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(7037..7056)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(7716..8576)
/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 8339..8358
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(8577..8681)
/gene="bla"
/label=AmpR promoter
primer_bind complement(8760..8779)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
ORIGIN
1 gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg
61 accgacaatt gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta
121 cgggccagat atacgcgttg acattgatta ttgactagtt attaatagta atcaattacg
181 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
241 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
301 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
361 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat
421 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact
481 tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac
541 atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac
601 gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac
661 tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga
721 gctggtttag tgaaccgtca gatccgctag agatccgcgg ccgctaatac gactcactat
781 agggagagcc gccaccatga aacggacagc cgacggaagc gagttcgagt caccaaagaa
841 gaagcggaaa gtctctgaag tcgagtttag ccacgagtat tggatgaggc acgcactgac
901 cctggcaaag cgagcatggg atgaaagaga agtccccgtg ggcgccgtgc tggtgcacaa
961 caatagagtg atcggagagg gatggaacag gccaatcggc cgccacgacc ctaccgcaca
1021 cgcagagatc atggcactga ggcagggagg cctggtcatg cagaattacc gcctgatcga
1081 tgccaccctg tatgtgacac tggagccatg cgtgatgtgc gcaggagcaa tgatccacag
1141 caggatcgga agagtggtgt tcggagcacg ggacgccaag accggcgcag caggctccct
1201 gatggatgtg ctgcaccacc ccggcatgaa ccaccgggtg gagatcacag agggaatcct
1261 ggcagacgag tgcgccgccc tgctgagcga tttctttaga atgcggagac aggagatcaa
1321 ggcccagaag aaggcacaga gctccaccga ctctggagga tctagcggag gatcctctgg
1381 aagcgagaca ccaggcacaa gcgagtccgc cacaccagag agctccggcg gctcctccgg
1441 aggatcctct gaggtggagt tttcccacga gtactggatg agacatgccc tgaccctggc
1501 caagagggca cgcgatgaga gggaggtgcc tgtgggagcc gtgctggtgc tgaacaatag
1561 agtgatcggc gagggctgga acagagccat cggcctgcac gacccaacag cccatgccga
1621 aattatggcc ctgagacagg gcggcctggt catgcagaac tacagactga ttgacgccac
1681 cctgtacgtg acattcgagc cttgcgtgat gtgcgccggc gccatgatcc actctaggat
1741 cggccgcgtg gtgtttggcg tgaggaacgc aaaaaccggc gccgcaggct ccctgatgga
1801 cgtgctgcac taccccggca tgaatcaccg cgtcgaaatt accgagggaa tcctggcaga
1861 tgaatgtgcc gccctgctgt gctatttctt tcggatgcct agacaggtgt tcaatgctca
1921 gaagaaggcc cagagctcca ccgactccgg aggatctagc ggaggctcct ctggctctga
1981 gacacctggc acaagcgaga gcgcaacacc tgaaagcagc gggggcagca gcggggggtc
2041 agacaagaag tacagcatcg gcctggccat cggcaccaac tctgtgggct gggccgtgat
2101 caccgacgag tacaaggtgc ccagcaagaa attcaaggtg ctgggcaaca ccgaccggca
2161 cagcatcaag aagaacctga tcggagccct gctgttcgac agcggcgaaa cagccgaggc
2221 cacccggctg aagagaaccg ccagaagaag atacaccaga cggaagaacc ggatctgcta
2281 tctgcaagag atcttcagca acgagatggc caaggtggac gacagcttct tccacagact
2341 ggaagagtcc ttcctggtgg aagaggataa gaagcacgag cggcacccca tcttcggcaa
2401 catcgtggac gaggtggcct accacgagaa gtaccccacc atctaccacc tgagaaagaa
2461 actggtggac agcaccgaca aggccgacct gcggctgatc tatctggccc tggcccacat
2521 gatcaagttc cggggccact tcctgatcga gggcgacctg aaccccgaca acagcgacgt
2581 ggacaagctg ttcatccagc tggtgcagac ctacaaccag ctgttcgagg aaaaccccat
2641 caacgccagc ggcgtggacg ccaaggccat cctgtctgcc agactgagca agagcagacg
2701 gctggaaaat ctgatcgccc agctgcccgg cgagaagaag aatggcctgt tcggaaacct
2761 gattgccctg agcctgggcc tgacccccaa cttcaagagc aacttcgacc tggccgagga
2821 tgccaaactg cagctgagca aggacaccta cgacgacgac ctggacaacc tgctggccca
2881 gatcggcgac cagtacgccg acctgtttct ggccgccaag aacctgtccg acgccatcct
2941 gctgagcgac atcctgagag tgaacaccga gatcaccaag gcccccctga gcgcctctat
3001 gatcaagaga tacgacgagc accaccagga cctgaccctg ctgaaagctc tcgtgcggca
3061 gcagctgcct gagaagtaca aagagatttt cttcgaccag agcaagaacg gctacgccgg
3121 ctacattgac ggcggagcca gccaggaaga gttctacaag ttcatcaagc ccatcctgga
3181 aaagatggac ggcaccgagg aactgctcgt gaagctgaac agagaggacc tgctgcggaa
3241 gcagcggacc ttcgacaacg gcagcatccc ccaccagatc cacctgggag agctgcacgc
3301 cattctgcgg cggcaggaag atttttaccc attcctgaag gacaaccggg aaaagatcga
3361 gaagatcctg accttccgca tcccctacta cgtgggccct ctggccaggg gaaacagcag
3421 attcgcctgg atgaccagaa agagcgagga aaccatcacc ccctggaact tcgaggaagt
3481 ggtggacaag ggcgcttccg cccagagctt catcgagcgg atgaccaact tcgataagaa
3541 cctgcccaac gagaaggtgc tgcccaagca cagcctgctg tacgagtact tcaccgtgta
3601 taacgagctg accaaagtga aatacgtgac cgagggaatg agaaagcccg ccttcctgag
3661 cggcgagcag aaaaaggcca tcgtggacct gctgttcaag accaaccgga aagtgaccgt
3721 gaagcagctg aaagaggact acttcaagaa aatcgagtgc ttcgactccg tggaaatctc
3781 cggcgtggaa gatcggttca acgcctccct gggcacatac cacgatctgc tgaaaattat
3841 caaggacaag gacttcctgg acaatgagga aaacgaggac attctggaag atatcgtgct
3901 gaccctgaca ctgtttgagg acagagagat gatcgaggaa cggctgaaaa cctatgccca
3961 cctgttcgac gacaaagtga tgaagcagct gaagcggcgg agatacaccg gctggggcag
4021 gctgagccgg aagctgatca acggcatccg ggacaagcag tccggcaaga caatcctgga
4081 tttcctgaag tccgacggct tcgccaacag aaacttcatg cagctgatcc acgacgacag
4141 cctgaccttt aaagaggaca tccagaaagc ccaggtgtcc ggccagggcg atagcctgca
4201 cgagcacatt gccaatctgg ccggcagccc cgccattaag aagggcatcc tgcagacagt
4261 gaaggtggtg gacgagctcg tgaaagtgat gggccggcac aagcccgaga acatcgtgat
4321 cgaaatggcc agagagaacc agaccaccca gaagggacag aagaacagcc gcgagagaat
4381 gaagcggatc gaagagggca tcaaagagct gggcagccag atcctgaaag aacaccccgt
4441 ggaaaacacc cagctgcaga acgagaagct gtacctgtac tacctgcaga atgggcggga
4501 tatgtacgtg gaccaggaac tggacatcaa ccggctgtcc gactacgatg tggaccatat
4561 cgtgcctcag agctttctga aggacgactc catcgacaac aaggtgctga ccagaagcga
4621 caagaaccgg ggcaagagcg acaacgtgcc ctccgaagag gtcgtgaaga agatgaagaa
4681 ctactggcgg cagctgctga acgccaagct gattacccag agaaagttcg acaatctgac
4741 caaggccgag agaggcggcc tgagcgaact ggataaggcc ggcttcatca agagacagct
4801 ggtggaaacc cggcagatca caaagcacgt ggcacagatc ctggactccc ggatgaacac
4861 taagtacgac gagaatgaca agctgatccg ggaagtgaaa gtgatcaccc tgaagtccaa
4921 gctggtgtcc gatttccgga aggatttcca gttttacaaa gtgcgcgaga tcaacaacta
4981 ccaccacgcc cacgacgcct acctgaacgc cgtcgtggga accgccctga tcaaaaagta
5041 ccctaagctg gaaagcgagt tcgtgtacgg cgactacaag gtgtacgacg tgcggaagat
5101 gatcgccaag agcgagcagg aaatcggcaa ggctaccgcc aagtacttct tctacagcaa
5161 catcatgaac tttttcaaga ccgagattac cctggccaac ggcgagatcc ggaagcggcc
5221 tctgatcgag acaaacggcg aaaccgggga gatcgtgtgg gataagggcc gggattttgc
5281 caccgtgcgg aaagtgctga gcatgcccca agtgaatatc gtgaaaaaga ccgaggtgca
5341 gacaggcggc ttcagcaaag agtctatcct gcccaagagg aacagcgata agctgatcgc
5401 cagaaagaag gactgggacc ctaagaagta cggcggcttc gtgagcccca ccgtggccta
5461 ttctgtgctg gtggtggcca aagtggaaaa gggcaagtcc aagaaactga agagtgtgaa
5521 agagctgctg gggatcacca tcatggaaag aagcagcttc gagaagaatc ccatcgactt
5581 tctggaagcc aagggctaca aagaagtgaa aaaggacctg atcatcaagc tgcctaagta
5641 ctccctgttc gagctggaaa acggccggaa gagaatgctg gcctcagcca gagaactgca
5701 gaagggaaac gaactggccc tgccctccaa atatgtgaac ttcctgtacc tggccagcca
5761 ctatgagaag ctgaagggct cccccgagga taatgagcag aaacagctgt ttgtggaaca
5821 gcacaagcac tacctggacg agatcatcga gcagatcagc gagttctcca agagagtgat
5881 cctggccgac gctaatctgg acaaagtgct gtccgcctac aacaagcacc gggataagcc
5941 catcagagag caggccgaga atatcatcca cctgtttacc ctgaccaatc tgggagcccc
6001 tgccgccttc aagtactttg acaccaccat cgaccggaag cagtacagaa gcaccaaaga
6061 ggtgctggac gccaccctga tccaccagag catcaccggc ctgtacgaga cacggatcga
6121 cctgtctcag ctgggaggtg actctggcgg ctcaaaaaga accgccgacg gcagcgaatt
6181 cgagcccaag aagaagagga aagtctaacc ggtcatcatc accatcacca ttgagtttaa
6241 acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc
6301 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag
6361 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag
6421 gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct
6481 atggcttctg aggcggaaag aaccagctgg ggctcgatac cgtcgacctc tagctagagc
6541 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
6601 cacaacatac gagccggaag cataaagtgt aaagcctagg gtgcctaatg agtgagctaa
6661 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
6721 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
6781 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
6841 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
6901 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
6961 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
7021 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
7081 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
7141 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
7201 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
7261 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
7321 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
7381 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
7441 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
7501 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
7561 ttttctacgg ggtctgacac tcagtggaac gaaaactcac gttaagggat tttggtcatg
7621 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
7681 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
7741 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
7801 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
7861 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
7921 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
7981 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
8041 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
8101 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
8161 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
8221 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
8281 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
8341 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
8401 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
8461 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
8521 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
8581 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
8641 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
8701 ccacctgacg tcgacggatc gggagatcga tctcccgatc ccctagggtc gactctcagt
8761 acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg t
//
