| 质粒名称: | pAtU6-sgRNA |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 119775 |
| 存储实验室: | Caixia Gao |
|
载体骨架基本信息 |
|
| 载体名称: | pUC57 |
| 载体出品公司: | -- |
| 空载体大小: | 3152 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: | AtU6-SmR-sgRNA,AtU6-sgRNA |
| 物种来源: | -- |
| 插入大小: | 1914 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子: | AtU6 |
| 标签/融合蛋白: | -- |
| 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: | pAtU6-sgRNA was a gift from Caixia Gao (Addgene plasmid # 119775 ; http://n2t.net/addgene:119775 ; RRID:Addgene_119775) |
| 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
pAtU6sgRNA载体基本信息
pAtU6sgRNA载体图谱质粒图谱和多克隆位点信息
pAtU6sgRNA载体简介
pAtU6sgRNA载体序列
LOCUS pAtU6-sgRNA 4629 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Expression of a target sgRNA in potato.
ACCESSION .
VERSION .
KEYWORDS pAtU6-sgRNA
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4629)
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 4629)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..4629
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(29..51)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 151..170
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 364..386
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 378..395
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 379..395
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
CDS 1180..1971
/codon_start=1
/gene="aadA"
/product="aminoglycoside adenylyltransferase (Murphy,
1985)"
/label=SmR
/note="confers resistance to spectinomycin and
streptomycin"
/translation="MGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
misc_RNA 2123..2198
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
primer_bind complement(2408..2424)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2408..2424)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(2421..2443)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 2432..2448
/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(2456..2486)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2501..2522
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(2639..2656)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(2810..3398)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(2890..2909)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(3569..4429)
/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 4192..4211
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(4430..4534)
/gene="bla"
/label=AmpR promoter
primer_bind 4602..4620
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
ORIGIN
1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cgagctcggt acctcgcgaa
421 tgcatctaga tatcggatcc cgggcccgtc gactgcagag gcctgcatgc aagcttcgac
481 ttgccttccg cacaatacat catttcttct tagctttttt tcttcttctt cgttcataca
541 gttttttttt gtttatcagc ttacattttc ttgaaccgta gctttcgttt tcttcttttt
601 aactttccat tcggagtttt tgtatcttgt ttcatagttt gtcccaggat tagaatgatt
661 aggcatcgaa ccttcaagaa tttgattgaa taaaacatct tcattcttaa gatatgaaga
721 taatcttcaa aaggcccctg ggaatctgaa agaagagaag caggcccatt tatatgggaa
781 agaacaatag tatttcttat ataggcccat ttaagttgaa aacaatcttc aaaagtccca
841 catcgcttag ataagaaaac gaagctgagt ttatatacag ctagagtcga agtagtgatt
901 gggagaccaa cccagtggac ataagcctgt tcggttcgta agctgtaatg caagtagcgt
961 atgcgctcac gcaactggtc cagaaccttg accgaacgca gcggtggtaa cggcgcagtg
1021 gcggttttca tggcttgtta tgactgtttt tttggggtac agtctatgcc tcgggcatcc
1081 aagcagcaag cgcgttacgc cgtgggtcga tgtttgatgt tatggagcag caacgatgtt
1141 acgcagcagg gcagtcgccc taaaacaaag ttaaacatca tgggggaagc ggtgatcgcc
1201 gaagtatcga ctcaactatc agaggtagtt ggcgtcatcg agcgccatct cgaaccgacg
1261 ttgctggccg tacatttgta cggctccgca gtggatggcg gcctgaagcc acacagtgat
1321 attgatttgc tggttacggt gaccgtaagg cttgatgaaa caacgcggcg agctttgatc
1381 aacgaccttt tggaaacttc ggcttcccct ggagagagcg agattctccg cgctgtagaa
1441 gtcaccattg ttgtgcacga cgacatcatt ccgtggcgtt atccagctaa gcgcgaactg
1501 caatttggag aatggcagcg caatgacatt cttgcaggta tcttcgagcc agccacgatc
1561 gacattgatc tggctatctt gctgacaaaa gcaagagaac atagcgttgc cttggtaggt
1621 ccagcggcgg aggaactctt tgatccggtt cctgaacagg atctatttga ggcgctaaat
1681 gaaaccttaa cgctatggaa ctcgccgccc gactgggctg gcgatgagcg aaatgtagtg
1741 cttacgttgt cccgcatttg gtacagcgca gtaaccggca aaatcgcgcc gaaggatgtc
1801 gctgccgact gggcaatgga gcgcctgccg gcccagtatc agcccgtcat acttgaagct
1861 agacaggctt atcttggaca agaagaagat cgcttggcct cgcgcgcaga tcagttggaa
1921 gaatttgtcc actacgtgaa aggcgagatc accaaggtag tcggcaaata atgtctagct
1981 agaaattcgt tcaagccgac gccgcttcgc ggcgcggctt aactcaagcg ttagatgcac
2041 taagcacata attgctcaca gccaaactat caggtcaagt ctgcttttat tatttttaag
2101 cgtgcataat aagccggtct cggttttaga gctagaaata gcaagttaaa ataaggctag
2161 tccgttatca acttgaaaaa gtggcaccga gtcggtgctt ttttttgcaa aattttccag
2221 atcgatttct tcttcctctg ttcttcggcg ttcaatttct ggggttttct cttcgttttc
2281 tgtaactgaa acctaaaatt tgacctaaaa aaaatctcaa ataatatgat tcagtggttt
2341 tgtacttttc agttagttga gttttgcagt tccgatgaga taaaccaata agcttggcgt
2401 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
2461 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
2521 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
2581 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
2641 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
2701 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
2761 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
2821 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
2881 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
2941 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
3001 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
3061 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
3121 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
3181 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
3241 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
3301 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
3361 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
3421 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
3481 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
3541 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
3601 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
3661 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gaaccacgct
3721 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
3781 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
3841 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
3901 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
3961 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
4021 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
4081 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
4141 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
4201 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
4261 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
4321 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
4381 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
4441 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
4501 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
4561 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
4621 cctttcgtc
//
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