| 质粒名称: | p35S-A3A-PBE |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 119773 |
| 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: | -- |
| 空载体大小: | 4919 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 植物表达 |
| 筛选标记: | -- |
| 载体抗性: | 氨苄霉素100 μg/mL |
| 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 |
| 物种来源: | -- |
| 插入大小: | 5106 bp |
| 基因突变: | nCas9 has a mutation D10A in wild type SpCas9 |
| GenBank编号 | -- |
| 启动子: | CaMv 2x35S |
| 标签/融合蛋白: | NLS (flanking UGI) |
| 克隆方法: | Gibson克隆 |
| 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: | p35S-A3A-PBE was a gift from Caixia Gao (Addgene plasmid # 119773 ; http://n2t.net/addgene:119773 ; RRID:Addgene_119773) |
| 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
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 p35S-A3A-PBE 8827 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Conversion of C to T in potato.
ACCESSION .
VERSION .
KEYWORDS p35S-A3A-PBE
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8827)
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 8827)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..8827
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 401..746
/label=CaMV 35S promoter
/note="strong constitutive promoter from cauliflower mosaic
virus"
primer_bind 689..709
/label=35S promoter
/note="CaMV 35S promoter, forward primer"
CDS 5521..5568
/codon_start=1
/product="bipartite nuclear localization signal from
nucleoplasmin"
/label=nucleoplasmin NLS
/translation="KRPAATKKAGQAKKKK"
CDS 5590..5838
/codon_start=1
/product="uracil-DNA glycosylase inhibitor from a Bacillus
subtilis bacteriophage (Mol et al., 1995)"
/label=UGI
/translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
CDS 5851..5871
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
polyA_signal 5945..6121
/label=CaMV poly(A) signal
/note="cauliflower mosaic virus polyadenylation signal"
primer_bind complement(6675..6691)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(6675..6691)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(6688..6710)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 6699..6715
/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(6723..6753)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 6768..6789
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(6906..6923)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(7077..7665)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(7157..7176)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(7836..8696)
/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 8459..8478
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(8697..8801)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 gagctcggta cccctactcc aaaaatgtca aagatacagt ctcagaagac caaagggcta
61 ttgagacttt tcaacaaagg gtaatttcgg gaaacctcct cggattccat tgcccagcta
121 tctgtcactt catcgaaagg acagtagaaa aggaaggtgg ctcctacaaa tgccatcatt
181 gcgataaagg aaaggctatc attcaagatg cctctgccga cagtggtccc aaagatggac
241 ccccacccac gaggagcatc gtggaaaaag aagacgttcc aaccacgtct tcaaagcaag
301 tggattgatg tgacatctcc actgacgtaa gggatgacgc acaatcccac ccctactcca
361 aaaatgtcaa agatacagtc tcagaagacc aaagggctat tgagactttt caacaaaggg
421 taatttcggg aaacctcctc ggattccatt gcccagctat ctgtcacttc atcgaaagga
481 cagtagaaaa ggaaggtggc tcctacaaat gccatcattg cgataaagga aaggctatca
541 ttcaagatgc ctctgccgac agtggtccca aagatggacc cccacccacg aggagcatcg
601 tggaaaaaga agacgttcca accacgtctt caaagcaagt ggattgatgt gacatctcca
661 ctgacgtaag ggatgacgca caatcccact atccttcgca agacccttcc tctatataag
721 gaagttcatt tcatttggag aggacagccc aagcttgtcg acggatccat ggaggccagc
781 ccggctagcg gcccaaggca tctcatggac ccgcacatct tcaccagcaa cttcaacaac
841 ggcatcggca ggcacaagac ctacttgtgc tacgaggtgg agaggctcga caacggaacc
901 tccgtgaaga tggaccaaca cagggggttc ctccacaacc aagccaagaa cctcctctgc
961 ggcttctacg gcaggcacgc cgagttgagg ttcctcgact tggtgccatc cctccaactc
1021 gatccagccc aaatctaccg cgtgacctgg ttcatctcct ggtccccatg cttctcctgg
1081 ggttgcgccg gcgaggttcg ggctttcctc caagaaaaca cccacgtccg cctccgcatt
1141 ttcgccgcca ggatctatga ttacgaccct ctctacaagg aggccctcca gatgctgcgg
1201 gacgccggtg ctcaggtgag tatcatgacc tacgacgagt tcaagcactg ctgggacacc
1261 ttcgttgacc accagggctg cccattccaa ccatgggacg gtctggatga acacagccaa
1321 gccttgtccg gcaggctccg ggccatcctc caaaaccagg ggaactccgg gagcgagacg
1381 ccaggcacct ccgagtcggc caccccagaa tctcttaagg acaagaagta ctcgatcggc
1441 ctcgccatcg ggacgaactc agttggctgg gccgtgatca ccgacgagta caaggtgccc
1501 tctaagaagt tcaaggtcct ggggaacacc gaccgccatt ccatcaagaa gaacctcatc
1561 ggcgctctcc tgttcgacag cggggagacc gctgaggcta cgaggctcaa gagaaccgct
1621 aggcgccggt acacgagaag gaagaacagg atctgctacc tccaagagat tttctccaac
1681 gagatggcca aggttgacga ttcattcttc caccgcctgg aggagtcttt cctcgtggag
1741 gaggataaga agcacgagcg gcatcccatc ttcggcaaca tcgtggacga ggttgcctac
1801 cacgagaagt accctacgat ctaccatctg cggaagaagc tcgtggactc caccgataag
1861 gcggacctca gactgatcta cctcgctctg gcccacatga tcaagttccg cggccatttc
1921 ctgatcgagg gggatctcaa cccagacaac agcgatgttg acaagctgtt catccaactc
1981 gtgcagacct acaaccaact cttcgaggag aacccgatca acgcctctgg cgtggacgcg
2041 aaggctatcc tgtccgcgag gctctcgaag tccaggaggc tggagaacct gatcgctcag
2101 ctcccaggcg agaagaagaa cggcctgttc gggaacctca tcgctctcag cctggggctc
2161 accccgaact tcaagtcgaa cttcgatctc gctgaggacg ccaagctgca actctccaag
2221 gacacctacg acgatgacct cgataacctc ctggcccaga tcggcgatca atacgcggac
2281 ctgttcctcg ctgccaagaa cctgtcggac gccatcctcc tgtcagatat cctccgcgtg
2341 aacaccgaga tcacgaaggc tccactctct gcctccatga tcaagcgcta cgacgagcac
2401 catcaggatc tgaccctcct gaaggcgctg gtccgccaac agctcccgga gaagtacaag
2461 gagattttct tcgatcagtc gaagaacggc tacgctgggt acatcgacgg cggggcctca
2521 caagaggagt tctacaagtt catcaagcca atcctggaga agatggacgg cacggaggag
2581 ctcctggtga agctcaacag ggaggacctc ctgcggaagc agagaacctt cgataacggc
2641 agcatccccc accaaatcca tctcggggag ctgcacgcca tcctgagaag gcaagaggac
2701 ttctaccctt tcctcaagga taaccgggag aagatcgaga agatcctgac cttcagaatc
2761 ccatactacg tcggccctct cgcgcggggg aactcaagat tcgcttggat gacccgcaag
2821 tctgaggaga ccatcacgcc gtggaacttc gaggaggtgg tggacaaggg cgctagcgct
2881 cagtcgttca tcgagaggat gaccaacttc gacaagaacc tgcccaacga gaaggtgctc
2941 cctaagcact cgctcctgta cgagtacttc accgtctaca acgagctcac gaaggtgaag
3001 tacgtcaccg agggcatgcg caagccagcg ttcctgtccg gggagcagaa gaaggctatc
3061 gtggacctcc tgttcaagac caaccggaag gtcacggtta agcaactcaa ggaggactac
3121 ttcaagaaga tcgagtgctt cgattcggtc gagatcagcg gcgttgagga ccgcttcaac
3181 gccagcctcg ggacctacca cgatctcctg aagatcatca aggataagga cttcctggac
3241 aacgaggaga acgaggatat cctggaggac atcgtgctga ccctcacgct gttcgaggac
3301 agggagatga tcgaggagcg cctgaagacg tacgcccatc tcttcgatga caaggtcatg
3361 aagcaactca agcgccggag atacaccggc tgggggaggc tgtcccgcaa gctcatcaac
3421 ggcatccggg acaagcagtc cgggaagacc atcctcgact tcctcaagag cgatggcttc
3481 gccaacagga acttcatgca actgatccac gatgacagcc tcaccttcaa ggaggatatc
3541 caaaaggctc aagtgagcgg ccagggggac tcgctgcacg agcatatcgc gaacctcgct
3601 ggctcccccg cgatcaagaa gggcatcctc cagaccgtga aggttgtgga cgagctcgtg
3661 aaggtcatgg gccggcacaa gcctgagaac atcgtcatcg agatggccag agagaaccaa
3721 accacgcaga aggggcaaaa gaactctagg gagcgcatga agcgcatcga ggagggcatc
3781 aaggagctgg ggtcccaaat cctcaaggag cacccagtgg agaacaccca actgcagaac
3841 gagaagctct acctgtacta cctccagaac ggcagggata tgtacgtgga ccaagagctg
3901 gatatcaacc gcctcagcga ttacgacgtc gatcatatcg ttccccagtc tttcctgaag
3961 gatgactcca tcgacaacaa ggtcctcacc aggtcggaca agaaccgcgg caagtcagat
4021 aacgttccat ctgaggaggt cgttaagaag atgaagaact actggaggca gctcctgaac
4081 gccaagctga tcacgcaaag gaagttcgac aacctcacca aggctgagag aggcgggctc
4141 tcagagctgg acaaggccgg cttcatcaag cggcagctgg tcgagaccag acaaatcacg
4201 aagcacgttg cgcaaatcct cgactctcgg atgaacacga agtacgatga gaacgacaag
4261 ctgatcaggg aggttaaggt gatcaccctg aagtctaagc tcgtctccga cttcaggaag
4321 gatttccagt tctacaaggt tcgcgagatc aacaactacc accatgccca tgacgcttac
4381 ctcaacgctg tggtcggcac cgctctgatc aagaagtacc caaagctgga gtccgagttc
4441 gtgtacgggg actacaaggt ttacgatgtg cgcaagatga tcgccaagtc ggagcaagag
4501 atcggcaagg ctaccgccaa gtacttcttc tactcaaaca tcatgaactt cttcaagacc
4561 gagatcacgc tggccaacgg cgagatccgg aagagaccgc tcatcgagac caacggcgag
4621 acgggggaga tcgtgtggga caagggcagg gatttcgcga ccgtccgcaa ggttctctcc
4681 atgccccagg tgaacatcgt caagaagacc gaggtccaaa cgggcgggtt ctcaaaggag
4741 tctatcctgc ctaagcggaa cagcgacaag ctcatcgcca gaaagaagga ctgggaccca
4801 aagaagtacg gcgggttcga cagccctacc gtggcctact cggtcctggt tgtggcgaag
4861 gttgagaagg gcaagtccaa gaagctcaag agcgtgaagg agctcctggg gatcaccatc
4921 atggagaggt ccagcttcga gaagaaccca atcgacttcc tggaggccaa gggctacaag
4981 gaggtgaaga aggacctgat catcaagctc ccgaagtact ctctcttcga gctggagaac
5041 ggcaggaaga gaatgctggc ttccgctggc gagctccaga aggggaacga gctcgcgctg
5101 ccaagcaagt acgtgaactt cctctacctg gcttcccact acgagaagct caagggcagc
5161 ccggaggaca acgagcaaaa gcagctgttc gtcgagcagc acaagcatta cctcgacgag
5221 atcatcgagc aaatctccga gttcagcaag cgcgtgatcc tcgccgacgc gaacctggat
5281 aaggtcctct ccgcctacaa caagcaccgg gacaagccca tcagagagca agcggagaac
5341 atcatccatc tcttcaccct gacgaacctc ggcgctcctg ctgctttcaa gtacttcgac
5401 accacgatcg atcggaagag atacacctcc acgaagaagg tcctggacgc gaccctcatc
5461 caccagtcga tcaccggcct gtacgagacg aggatcgacc tctcacaact cggcggggat
5521 aagagacccg cagcaaccaa gaaggcaggg caagcaaaga agaagaagac gcgtgactcc
5581 ggcggcagca ccaacctgtc cgacatcatc gagaaggaga cgggcaagca actcgtgatc
5641 caggagagca tcctcatgct gccagaggag gtggaggagg tcatcggcaa caagccagag
5701 tccgacatcc tggtgcacac cgcctacgac gagtccaccg acgagaacgt catgctcctg
5761 accagcgacg ccccagagta caagccatgg gccctcgtca tccaggacag caacggggag
5821 aacaagatca agatgctgtc gggggggagc ccaaagaaga agcggaaggt gtagtgtaca
5881 agtaacccgg gaattcggta cgctgaaatc accagtctct ctctacaaat ctatctctct
5941 ctattttctc cataaataat gtgtgagtag tttcccgata agggaaatta gggttcttat
6001 agggtttcgc tcatgtgttg agcatataag aaacccttag tatgtatttg tatttgtaaa
6061 atacttctat caataaaatt tctaattcct aaaaccaaaa tccagtacta aaatccagat
6121 ctcctaaagt ccctatagat ctttgtcgtg aatataaacc agacacgaga cgactaaacc
6181 tggagcccag acgccgttcg aagctagaag taccgcttag gcaggaggcc gttagggaaa
6241 agatgctaag gcagggttgg ttacgttgac tcccccgtag gtttggttta aatatgatga
6301 agtggacgga aggaaggagg aagacaagga aggataaggt tgcaggccct gtgcaaggta
6361 agaagatgga aatttgatag aggtacgcta ctatacttat actatacgct aagggaatgc
6421 ttgtatttat accctatacc ccctaataac cccttatcaa tttaagaaat aatccgcata
6481 agcccccgct taaaaattgg tatcagagcc atgaataggt ctatgaccaa aactcaagag
6541 gataaaacct caccaaaata cgaaagagtt cttaactcta aagataaaag atctttcaag
6601 atcaaaacta gttccctcac accggtgacg gggatcgcat gcgatatctc gagatctagc
6661 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
6721 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
6781 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
6841 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
6901 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
6961 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
7021 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
7081 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
7141 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
7201 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
7261 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
7321 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
7381 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
7441 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
7501 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
7561 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
7621 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
7681 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
7741 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
7801 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
7861 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
7921 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
7981 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
8041 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
8101 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
8161 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
8221 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
8281 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
8341 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
8401 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
8461 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
8521 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
8581 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
8641 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
8701 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
8761 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
8821 ccacctg
//
