| 质粒名称: | pYLsgRNA-AtU3d |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 66200 |
| 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-AtU3d was a gift from Yao-Guang Liu (Addgene plasmid # 66200 ; http://n2t.net/addgene:66200 ; RRID:Addgene_66200) |
| 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-AtU3d 2944 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION cloning of sgRNAs for expression in plants.
ACCESSION .
VERSION .
KEYWORDS pYLsgRNA-AtU3d
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 2944)
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 2944)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..2944
/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"
primer_bind complement(298..315)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(298..314)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(307..329)
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind complement(523..542)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 642..664
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(702..720)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 788..892
/gene="bla"
/label=AmpR promoter
CDS 893..1753
/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(1111..1130)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 1924..2512
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2413..2432
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 2666..2683
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 2800..2821
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 2836..2866
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2874..2890
/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 2879..2901
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 2898..2914
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 2898..2914
/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 gaataagctt atgatttctt
181 ttttcttacg aattttgcgt cccacatcgg taagcgagtg aagaaataac tgctttatat
241 atggctacaa agcaccattg gtcaagagac ctctgaagat aacatactaa gcttggcact
301 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
361 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc
421 ttcccaacag ttgcgcagcc tgaatggcga atggcgcctg atgcggtatt ttctccttac
481 gcatctgtgc ggtatttcac accgcatatg gtgcactctc agtacaatct gctctgatgc
541 cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
601 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
661 gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt
721 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
781 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
841 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
901 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
961 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
1021 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
1081 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
1141 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
1201 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
1261 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
1321 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
1381 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
1441 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
1501 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
1561 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
1621 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
1681 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
1741 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
1801 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
1861 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
1921 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
1981 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
2041 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
2101 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
2161 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
2221 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
2281 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
2341 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
2401 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
2461 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
2521 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
2581 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
2641 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
2701 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
2761 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca
2821 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag
2881 cggataacaa tttcacacag gaaacagcta tgaccatgat tacgaattcg agctcggtac
2941 ccgg
//
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