| 质粒名称: | pU3-gRNA |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 53063 |
| 存储实验室: | Caixia Gao |
|
载体骨架基本信息 |
|
| 载体名称: | pUC-T |
| 载体出品公司: | CWBIO |
| 空载体大小: | 2700bp |
| 完整质粒大小: | 3193bp |
| 载体类型: | CRISPR ; Plant expression |
| 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: | OsU3 promoter + gRNA scaffold |
| 物种来源: | rice U3 promoter |
| 插入大小: | 493bp |
| 启动子: | rice U3 |
| 克隆方法: | TOPO Cloning |
| 5’克隆位点: | AgeI(未破坏) |
| 3’克隆位点: | NotI (未破坏) |
| 5’测序引物 | M13F |
| 3’测序引物 | M13R |
|
文献引用方法 |
|
| 材料和方法部分: | pU3-gRNA was a gift from Caixia Gao (Addgene plasmid # 53063 ; http://n2t.net/addgene:53063 ; RRID:Addgene_53063) |
| 参考文献部分: | Targeted genome modification of crop plants using a CRISPR-Cas system. Shan Q, Wang Y, Li J, Zhang Y, Chen K, Liang Z, Zhang K, Liu J, Xi JJ, Qiu JL, Gao C. Nat Biotechnol. 2013 Aug;31(8):686-8. doi: 10.1038/nbt.2650. 10.1038/nbt.2650 PubMed 23929338 |
买家导航
pU3-gRNA载体质粒基本信息
pU3-gRNA质粒图谱载体图谱和pU3-gRNA载体序列质粒序列多克隆位点信息
pU3-gRNA质粒载体简介
pU3-gRNA质粒序列载体序列
LOCUS pU3-gRNA 3193 bp DNA circular SYN 14-APR-2025
DEFINITION Transcription of guide RNA (gRNA) in rice cells.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3193)
AUTHORS Shan Q, Wang Y, Li J, Zhang Y, Chen K, Liang Z, Zhang K, Liu J, Xi
JJ, Qiu JL, Gao C
TITLE Targeted genome modification of crop plants using a CRISPR-Cas
system.
JOURNAL Nat Biotechnol. 2013 Aug;31(8):686-8. doi: 10.1038/nbt.2650.
PUBMED 23929338
REFERENCE 2 (bases 1 to 3193)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 14, 2025 from SnapGene Server 8.0.1
https://www.biofeng.com
COMMENT SGRef: number: 1; type: "Journal Article"; doi: "10.1038/nbt";
journalName: "Nat Biotechnol"; date: "2013-08"; volume: "31"; issue:
"8"; pages: "686-8"
COMMENT Sequence Label: pU3-gRNA
FEATURES Location/Qualifiers
source 1..3193
/mol_type="other DNA"
/organism="synthetic DNA construct"
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"
promoter 427..807
/label=OsU3 promoter
/note="Oryza sativa (rice) snRNA U3 promoter"
misc_RNA 837..912
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
primer_bind complement(972..988)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(972..988)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(985..1007)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 996..1012
/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(join(1020..1026,1027..1044,1045..1050))
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 1065..1086
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(1203..1220)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(1374..1962)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(1454..1473)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(join(2133..2924,2925..2993))
/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 2756..2775
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(2994..3098)
/gene="bla"
/label=AmpR promoter
primer_bind 3166..3184
/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 ccagtgccaa gcttgcatgc ctgcaggtcg
421 acgattaagg aatctttaaa catacgaaca gatcacttaa agttcttctg aagcaactta
481 aagttatcag gcatgcatgg atcttggagg aatcagatgt gcagtcaggg accatagcac
541 aagacaggcg tcttctactg gtgctaccag caaatgctgg aagccgggaa cactgggtac
601 gtcggaaacc acgtgatgtg aagaagtaag ataaactgta ggagaaaagc atttcgtagt
661 gggccatgaa gcctttcagg acatgtattg cagtatgggc cggcccatta cgcaattgga
721 cgacaacaaa gactagtatt agtaccacct cggctatcca catagatcaa agctgattta
781 aaagagttgt gcagatgatc cgtggcagct cgcaggtgaa cacaacacct gcacacgttt
841 tagagctaga aatagcaagt taaaataagg ctagtccgtt atcaacttga aaaagtggca
901 ccgagtcggt gctttttttc cacataatct ctagaggatc cccgggtacc gagctcgaat
961 tcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
1021 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
1081 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
1141 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
1201 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
1261 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
1321 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
1381 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
1441 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
1501 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
1561 ctttctcaaa gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
1621 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
1681 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
1741 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
1801 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
1861 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
1921 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
1981 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
2041 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
2101 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
2161 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
2221 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
2281 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
2341 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
2401 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
2461 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
2521 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
2581 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
2641 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
2701 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
2761 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
2821 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
2881 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
2941 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
3001 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
3061 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
3121 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
3181 aggccctttc gtc
//
