| 质粒名称: | pYLsgRNA-OsU3m |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 66193 |
| 存储实验室: | Yao-Guang Liu |
|
载体骨架基本信息 |
|
| 载体名称: | based on pUC18 |
| 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: | 人细胞色素c氧化酶亚基VIII（human cytochrome C oxidase subunit VIII） |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 新霉素（G418） |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | -- |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | 人细胞色素c氧化酶亚基VIII（N端） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | AgeI(未破坏) |
| 3' 克隆位点: | NotI (未破坏) |
| 5' 测序引物 | CMV-F |
| 3' 测序引物 | SV40pA-R |
|
文献引用方法 |
|
| 材料和方法部分: | pYLsgRNA-OsU3m was a gift from Yao-Guang Liu (Addgene plasmid # 66193 ; http://n2t.net/addgene:66193 ; RRID:Addgene_66193) |
| 参考文献部分: | A robust CRISPR/Cas9 system for convenient, high-efficiency multiplex genome editing in monocot and dicot plants. Ma X, Zhang Q, Zhu Q, Liu W, Chen Y, Qiu R, Wang B, Yang Z, Li H, Lin Y, Xie Y, Shen R, Chen S, Wang Z, Chen Y, Guo J, Chen L, Zhao X, Dong Z, Liu YG. Mol Plant. 2015 Apr 24. pii: S1674-2052(15)00204-X. doi: 10.1016/j.molp.2015.04.007. 10.1016/j.molp.2015.04.007 PubMed 25917172 |
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 pYLsgRNA-OsU3m 3224 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION cloning of sgRNAs for expression in plants.
ACCESSION .
VERSION .
KEYWORDS pYLsgRNA-OsU3m
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3224)
AUTHORS Ma X, Zhang Q, Zhu Q, Liu W, Chen Y, Qiu R, Wang B, Yang Z, Li H,
Lin Y, Xie Y, Shen R, Chen S, Wang Z, Chen Y, Guo J, Chen L, Zhao X,
Dong Z, Liu YG
TITLE A robust CRISPR/Cas9 system for convenient, high-efficiency
multiplex genome editing in monocot and dicot plants.
JOURNAL Mol Plant. 2015 Apr 24. pii: S1674-2052(15)00204-X. doi:
10.1016/j.molp.2015.04.007.
PUBMED 25917172
REFERENCE 2 (bases 1 to 3224)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..3224
/organism="synthetic DNA construct"
/mol_type="other DNA"
misc_RNA 20..95
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
promoter 162..544
/label=OsU3 promoter
/note="Oryza sativa (rice) snRNA U3 promoter"
primer_bind complement(578..595)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(578..594)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(587..609)
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind complement(803..822)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 922..944
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(982..1000)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 1068..1172
/gene="bla"
/label=AmpR promoter
CDS 1173..2033
/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 complement(1391..1410)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 2204..2792
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2693..2712
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 2946..2963
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 3080..3101
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 3116..3146
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3154..3170
/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)."
primer_bind 3159..3181
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 3178..3194
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 3178..3194
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
ORIGIN
1 ggatccagcg tgggtctcgg ttttagagct agaaatagca agttaaaata aggctagtcc
61 gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt ttcaagagct tggagtggat
121 ggaattttcc tccgttttac ctgtggaatc ggcagcaaag gaaggaatct ttaaacatac
181 gaacagatca cttaaagttc ttctgaagca acttaaagtt atcaggcatg catggatctt
241 ggaggaatca gatgtgcagt cagggaccat agcacaagac aggcgtcttc tactggtgct
301 accagcaaat gctggaagcc gggaacactg ggtacgttgg aaaccacgtg tgatgtgaag
361 gagtaagata aactgtagga gaaaagcatt tcgtagtggg ccatgaagcc tttcaggaca
421 tgtattgcag tatgggccgg cccattacgc aattggacga caacaaaggc tagtattagt
481 accacctcgg ctatccacat agatcaaagc tggtttaaaa gagttgtgca gatgatccgt
541 ggcaagagac ctctgaagat aacatactaa gcttggcact ggccgtcgtt ttacaacgtc
601 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg
661 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
721 tgaatggcga atggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
781 accgcatatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc
841 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
901 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
961 cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
1021 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
1081 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
1141 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
1201 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
1261 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
1321 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
1381 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
1441 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
1501 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
1561 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
1621 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
1681 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
1741 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
1801 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
1861 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
1921 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
1981 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
2041 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
2101 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
2161 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
2221 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
2281 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
2341 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
2401 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
2461 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
2521 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
2581 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
2641 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
2701 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
2761 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
2821 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
2881 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
2941 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc
3001 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
3061 gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac
3121 actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag
3181 gaaacagcta tgaccatgat tacgaattcg agctcggtac ccgg
//
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