| 质粒名称: | A3A-PBE |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 119768 |
| 存储实验室: | Caixia Gao |
|
载体骨架基本信息 |
|
| 载体名称: | pJIT163 |
| 载体出品公司: | Clontech |
| 空载体大小: | 4919bp |
| 完整质粒大小: | 10080bp |
| 载体类型: | 植物表达 |
| 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: | APOBEC3A-nCas9-NLS-UGI-NLS、APOBEC3A-nCas9-UGI |
| 插入大小: | 5106bp |
| 基因突变: | nCas9 has a mutation D10A and E945K in wild type SpCas9 |
| 启动子: | maize Ubiquitin-1 |
| 标签/融合蛋白: | NLS (flanking UGI) |
| 克隆方法: | Gateway Cloning |
| 5’测序引物 | N/A |
|
文献引用方法 |
|
| 材料和方法部分: | A3A-PBE was a gift from Caixia Gao (Addgene plasmid # 119768 ; http://n2t.net/addgene:119768 ; RRID:Addgene_119768) |
| 参考文献部分: | Efficient C-to-T base editing in plants using a fusion of nCas9 and human APOBEC3A. Zong Y, Song Q, Li C, Jin S, Zhang D, Wang Y, Qiu JL, Gao C. Nat Biotechnol. 2018 Oct 1. pii: nbt.4261. doi: 10.1038/nbt.4261. 10.1038/nbt.4261 PubMed 30272679 |
买家导航
A3A-PBE载体质粒基本信息
A3A-PBE质粒图谱载体图谱和A3A-PBE载体序列质粒序列多克隆位点信息
A3A-PBE质粒载体简介
A3A-PBE质粒序列载体序列
LOCUS A3A-PBE 10080 bp DNA circular SYN 26-AUG-2024
DEFINITION Conversion of C to T in wheat plants or monocotyledons protoplasts .
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10080)
AUTHORS Zong Y, Song Q, Li C, Jin S, Zhang D, Wang Y, Qiu JL, Gao C
TITLE Efficient C-to-T base editing in plants using a fusion of nCas9 and
human APOBEC3A.
JOURNAL Nat Biotechnol. 2018 Oct 1. pii: nbt.4261. doi: 10.1038/nbt.4261.
PUBMED 30272679
REFERENCE 2 (bases 1 to 10080)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.biofeng.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat
Biotechnol. 2018 Oct 1. pii: nbt.4261. doi: 10.1038/nbt.4261."
FEATURES Location/Qualifiers
source 1..10080
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS 6770..6817
/codon_start=1
/product="bipartite nuclear localization signal from
nucleoplasmin"
/label=nucleoplasmin NLS
/translation="KRPAATKKAGQAKKKK"
CDS 6839..7087
/codon_start=1
/product="uracil-DNA glycosylase inhibitor from a Bacillus
subtilis bacteriophage (Mol et al., 1995)"
/label=UGI
/translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
CDS 7100..7120
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
polyA_signal 7194..7370
/label=CaMV poly(A) signal
/note="cauliflower mosaic virus polyadenylation signal"
primer_bind complement(7924..7940)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(7924..7940)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(7937..7959)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 7948..7964
/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(7972..8002)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 8017..8038
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(8155..8172)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(8326..8914)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(8406..8425)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(9085..9945)
/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 9708..9727
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(9946..10050)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 gagctcggta cctgacccgg tcgtgcccct ctctagagat aatgagcatt gcatgtctaa
61 gttataaaaa attaccacat attttttttg tcacacttgt ttgaagtgca gtttatctat
121 ctttatacat atatttaaac tttactctac gaataatata atctatagta ctacaataat
181 atcagtgttt tagagaatca tataaatgaa cagttagaca tggtctaaag gacaattgag
241 tattttgaca acaggactct acagttttat ctttttagtg tgcatgtgtt ctcctttttt
301 tttgcaaata gcttcaccta tataatactt catccatttt attagtacat ccatttaggg
361 tttagggtta atggttttta tagactaatt tttttagtac atctatttta ttctatttta
421 gcctctaaat taagaaaact aaaactctat tttagttttt ttatttaata atttagatat
481 aaaatagaat aaaataaagt gactaaaaat taaacaaata ccctttaaga aattaaaaaa
541 actaaggaaa catttttctt gtttcgagta gataatgcca gcctgttaaa cgccgtcgac
601 gagtctaacg gacaccaacc agcgaaccag cagcgtcgcg tcgggccaag cgaagcagac
661 ggcacggcat ctctgtcgct gcctctggac ccctctcgat cgagagttcc gctccaccgt
721 tggacttgct ccgctgtcgg catccagaaa ttgcgtggcg gagcggcaga cgtgagccgg
781 cacggcaggc ggcctcctcc tcctctcacg gcaccggcag ctacggggga ttcctttccc
841 accgctcctt cgctttccct tcctcgcccg ccgtaataaa tagacacccc ctccacaccc
901 tctttcccca acctcgtgtt gttcggagcg cacacacaca caaccagatc tcccccaaat
961 ccacccgtcg gcacctccgc ttcaaggtac gccgctcgtc ctcccccccc ccccctctct
1021 accttctcta gatcggcgtt ccggtccatg gttagggccc ggtagttcta cttctgttca
1081 tgtttgtgtt agatccgtgt ttgtgttaga tccgtgctgc tagcgttcgt acacggatgc
1141 gacctgtacg tcagacacgt tctgattgct aacttgccag tgtttctctt tggggaatcc
1201 tgggatggct ctagccgttc cgcagacggg atcgatttca tgattttttt tgtttcgttg
1261 catagggttt ggtttgccct tttcctttat ttcaatatat gccgtgcact tgtttgtcgg
1321 gtcatctttt catgcttttt tttgtcttgg ttgtgatgat gtggtctggt tgggcggtcg
1381 ttctagatcg gagtagaatt aattctgttt caaactacct ggtggattta ttaattttgg
1441 atctgtatgt gtgtgccata catattcata gttacgaatt gaagatgatg gatggaaata
1501 tcgatctagg ataggtatac atgttgatgc gggttttact gatgcatata cagagatgct
1561 ttttgttcgc ttggttgtga tgatgtggtg tggttgggcg gtcgttcatt cgttctagat
1621 cggagtagaa tactgtttca aactacctgg tgtatttatt aattttggaa ctgtatgtgt
1681 gtgtcataca tcttcatagt tacgagttta agatggatgg aaatatcgat ctaggatagg
1741 tatacatgtt gatgtgggtt ttactgatgc atatacatga tggcatatgc agcatctatt
1801 catatgctct aaccttgagt acctatctat tataataaac aagtatgttt tataattatt
1861 ttgatcttga tatacttgga tgatggcata tgcagcagct atatgtggat ttttttagcc
1921 ctgccttcat acgctattta tttgcttggt actgtttctt ttgtcgatgc tcaccctgtt
1981 gtttggtgtt acttctgcaa agcttgtcga cggatccatg gaggccagcc cggctagcgg
2041 cccaaggcat ctcatggacc cgcacatctt caccagcaac ttcaacaacg gcatcggcag
2101 gcacaagacc tacttgtgct acgaggtgga gaggctcgac aacggaacct ccgtgaagat
2161 ggaccaacac agggggttcc tccacaacca agccaagaac ctcctctgcg gcttctacgg
2221 caggcacgcc gagttgaggt tcctcgactt ggtgccatcc ctccaactcg atccagccca
2281 aatctaccgc gtgacctggt tcatctcctg gtccccatgc ttctcctggg gttgcgccgg
2341 cgaggttcgg gctttcctcc aagaaaacac ccacgtccgc ctccgcattt tcgccgccag
2401 gatctatgat tacgaccctc tctacaagga ggccctccag atgctgcggg acgccggtgc
2461 tcaggtgagt atcatgacct acgacgagtt caagcactgc tgggacacct tcgttgacca
2521 ccagggctgc ccattccaac catgggacgg tctggatgaa cacagccaag ccttgtccgg
2581 caggctccgg gccatcctcc aaaaccaggg gaactccggg agcgagacgc caggcacctc
2641 cgagtcggcc accccagaat ctcttaagga caagaagtac tcgatcggcc tcgccatcgg
2701 gacgaactca gttggctggg ccgtgatcac cgacgagtac aaggtgccct ctaagaagtt
2761 caaggtcctg gggaacaccg accgccattc catcaagaag aacctcatcg gcgctctcct
2821 gttcgacagc ggggagaccg ctgaggctac gaggctcaag agaaccgcta ggcgccggta
2881 cacgagaagg aagaacagga tctgctacct ccaagagatt ttctccaacg agatggccaa
2941 ggttgacgat tcattcttcc accgcctgga ggagtctttc ctcgtggagg aggataagaa
3001 gcacgagcgg catcccatct tcggcaacat cgtggacgag gttgcctacc acgagaagta
3061 ccctacgatc taccatctgc ggaagaagct cgtggactcc accgataagg cggacctcag
3121 actgatctac ctcgctctgg cccacatgat caagttccgc ggccatttcc tgatcgaggg
3181 ggatctcaac ccagacaaca gcgatgttga caagctgttc atccaactcg tgcagaccta
3241 caaccaactc ttcgaggaga acccgatcaa cgcctctggc gtggacgcga aggctatcct
3301 gtccgcgagg ctctcgaagt ccaggaggct ggagaacctg atcgctcagc tcccaggcga
3361 gaagaagaac ggcctgttcg ggaacctcat cgctctcagc ctggggctca ccccgaactt
3421 caagtcgaac ttcgatctcg ctgaggacgc caagctgcaa ctctccaagg acacctacga
3481 cgatgacctc gataacctcc tggcccagat cggcgatcaa tacgcggacc tgttcctcgc
3541 tgccaagaac ctgtcggacg ccatcctcct gtcagatatc ctccgcgtga acaccgagat
3601 cacgaaggct ccactctctg cctccatgat caagcgctac gacgagcacc atcaggatct
3661 gaccctcctg aaggcgctgg tccgccaaca gctcccggag aagtacaagg agattttctt
3721 cgatcagtcg aagaacggct acgctgggta catcgacggc ggggcctcac aagaggagtt
3781 ctacaagttc atcaagccaa tcctggagaa gatggacggc acggaggagc tcctggtgaa
3841 gctcaacagg gaggacctcc tgcggaagca gagaaccttc gataacggca gcatccccca
3901 ccaaatccat ctcggggagc tgcacgccat cctgagaagg caagaggact tctacccttt
3961 cctcaaggat aaccgggaga agatcgagaa gatcctgacc ttcagaatcc catactacgt
4021 cggccctctc gcgcggggga actcaagatt cgcttggatg acccgcaagt ctgaggagac
4081 catcacgccg tggaacttcg aggaggtggt ggacaagggc gctagcgctc agtcgttcat
4141 cgagaggatg accaacttcg acaagaacct gcccaacgag aaggtgctcc ctaagcactc
4201 gctcctgtac gagtacttca ccgtctacaa cgagctcacg aaggtgaagt acgtcaccga
4261 gggcatgcgc aagccagcgt tcctgtccgg ggagcagaag aaggctatcg tggacctcct
4321 gttcaagacc aaccggaagg tcacggttaa gcaactcaag gaggactact tcaagaagat
4381 cgagtgcttc gattcggtcg agatcagcgg cgttgaggac cgcttcaacg ccagcctcgg
4441 gacctaccac gatctcctga agatcatcaa ggataaggac ttcctggaca acgaggagaa
4501 cgaggatatc ctggaggaca tcgtgctgac cctcacgctg ttcgaggaca gggagatgat
4561 cgaggagcgc ctgaagacgt acgcccatct cttcgatgac aaggtcatga agcaactcaa
4621 gcgccggaga tacaccggct gggggaggct gtcccgcaag ctcatcaacg gcatccggga
4681 caagcagtcc gggaagacca tcctcgactt cctcaagagc gatggcttcg ccaacaggaa
4741 cttcatgcaa ctgatccacg atgacagcct caccttcaag gaggatatcc aaaaggctca
4801 agtgagcggc cagggggact cgctgcacga gcatatcgcg aacctcgctg gctcccccgc
4861 gatcaagaag ggcatcctcc agaccgtgaa ggttgtggac gagctcgtga aggtcatggg
4921 ccggcacaag cctgagaaca tcgtcatcga gatggccaga gagaaccaaa ccacgcagaa
4981 ggggcaaaag aactctaggg agcgcatgaa gcgcatcgag gagggcatca aggagctggg
5041 gtcccaaatc ctcaaggagc acccagtgga gaacacccaa ctgcagaacg agaagctcta
5101 cctgtactac ctccagaacg gcagggatat gtacgtggac caagagctgg atatcaaccg
5161 cctcagcgat tacgacgtcg atcatatcgt tccccagtct ttcctgaagg atgactccat
5221 cgacaacaag gtcctcacca ggtcggacaa gaaccgcggc aagtcagata acgttccatc
5281 tgaggaggtc gttaagaaga tgaagaacta ctggaggcag ctcctgaacg ccaagctgat
5341 cacgcaaagg aagttcgaca acctcaccaa ggctgagaga ggcgggctct cagagctgga
5401 caaggccggc ttcatcaagc ggcagctggt cgagaccaga caaatcacga agcacgttgc
5461 gcaaatcctc gactctcgga tgaacacgaa gtacgataag aacgacaagc tgatcaggga
5521 ggttaaggtg atcaccctga agtctaagct cgtctccgac ttcaggaagg atttccagtt
5581 ctacaaggtt cgcgagatca acaactacca ccatgcccat gacgcttacc tcaacgctgt
5641 ggtcggcacc gctctgatca agaagtaccc aaagctggag tccgagttcg tgtacgggga
5701 ctacaaggtt tacgatgtgc gcaagatgat cgccaagtcg gagcaagaga tcggcaaggc
5761 taccgccaag tacttcttct actcaaacat catgaacttc ttcaagaccg agatcacgct
5821 ggccaacggc gagatccgga agagaccgct catcgagacc aacggcgaga cgggggagat
5881 cgtgtgggac aagggcaggg atttcgcgac cgtccgcaag gttctctcca tgccccaggt
5941 gaacatcgtc aagaagaccg aggtccaaac gggcgggttc tcaaaggagt ctatcctgcc
6001 taagcggaac agcgacaagc tcatcgccag aaagaaggac tgggacccaa agaagtacgg
6061 cgggttcgac agccctaccg tggcctactc ggtcctggtt gtggcgaagg ttgagaaggg
6121 caagtccaag aagctcaaga gcgtgaagga gctcctgggg atcaccatca tggagaggtc
6181 cagcttcgag aagaacccaa tcgacttcct ggaggccaag ggctacaagg aggtgaagaa
6241 ggacctgatc atcaagctcc cgaagtactc tctcttcgag ctggagaacg gcaggaagag
6301 aatgctggct tccgctggcg agctccagaa ggggaacgag ctcgcgctgc caagcaagta
6361 cgtgaacttc ctctacctgg cttcccacta cgagaagctc aagggcagcc cggaggacaa
6421 cgagcaaaag cagctgttcg tcgagcagca caagcattac ctcgacgaga tcatcgagca
6481 aatctccgag ttcagcaagc gcgtgatcct cgccgacgcg aacctggata aggtcctctc
6541 cgcctacaac aagcaccggg acaagcccat cagagagcaa gcggagaaca tcatccatct
6601 cttcaccctg acgaacctcg gcgctcctgc tgctttcaag tacttcgaca ccacgatcga
6661 tcggaagaga tacacctcca cgaaggaggt cctggacgcg accctcatcc accagtcgat
6721 caccggcctg tacgagacga ggatcgacct ctcacaactc ggcggggata agagacccgc
6781 agcaaccaag aaggcagggc aagcaaagaa gaagaagacg cgtgactccg gcggcagcac
6841 caacctgtcc gacatcatcg agaaggagac gggcaagcaa ctcgtgatcc aggagagcat
6901 cctcatgctg ccagaggagg tggaggaggt catcggcaac aagccagagt ccgacatcct
6961 ggtgcacacc gcctacgacg agtccaccga cgagaacgtc atgctcctga ccagcgacgc
7021 cccagagtac aagccatggg ccctcgtcat ccaggacagc aacggggaga acaagatcaa
7081 gatgctgtcg ggggggagcc caaagaagaa gcggaaggtg tagtgtacaa gtaacccggg
7141 aattcggtac gctgaaatca ccagtctctc tctacaaatc tatctctctc tattttctcc
7201 ataaataatg tgtgagtagt ttcccgataa gggaaattag ggttcttata gggtttcgct
7261 catgtgttga gcatataaga aacccttagt atgtatttgt atttgtaaaa tacttctatc
7321 aataaaattt ctaattccta aaaccaaaat ccagtactaa aatccagatc tcctaaagtc
7381 cctatagatc tttgtcgtga atataaacca gacacgagac gactaaacct ggagcccaga
7441 cgccgttcga agctagaagt accgcttagg caggaggccg ttagggaaaa gatgctaagg
7501 cagggttggt tacgttgact cccccgtagg tttggtttaa atatgatgaa gtggacggaa
7561 ggaaggagga agacaaggaa ggataaggtt gcaggccctg tgcaaggtaa gaagatggaa
7621 atttgataga ggtacgctac tatacttata ctatacgcta agggaatgct tgtatttata
7681 ccctataccc cctaataacc ccttatcaat ttaagaaata atccgcataa gcccccgctt
7741 aaaaattggt atcagagcca tgaataggtc tatgaccaaa actcaagagg ataaaacctc
7801 accaaaatac gaaagagttc ttaactctaa agataaaaga tctttcaaga tcaaaactag
7861 ttccctcaca ccggtgacgg ggatcgcatg cgatatctcg agatctagct tggcgtaatc
7921 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
7981 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
8041 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
8101 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
8161 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
8221 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
8281 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
8341 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
8401 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
8461 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
8521 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
8581 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
8641 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
8701 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
8761 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
8821 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
8881 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
8941 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
9001 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
9061 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
9121 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
9181 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
9241 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
9301 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
9361 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
9421 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
9481 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
9541 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
9601 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
9661 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
9721 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
9781 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
9841 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
9901 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
9961 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
10021 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgackt
//
