质粒名称: | pYLsgRNA-AtU3b/LacZ |
---|---|
出品公司: | 爱迪基因 |
目录编号: | 66199 |
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: | -- |
-- | |
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-AtU3b/LacZ was a gift from Yao-Guang Liu (Addgene plasmid # 66199 ; http://n2t.net/addgene:66199 ; RRID:Addgene_66199) |
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-AtU3b/LacZ 3369 bp ds-DNA circular SYN 06-AUG-2018 DEFINITION cloning of sgRNAs for expression in plants. ACCESSION . VERSION . KEYWORDS pYLsgRNA-AtU3b/LacZ SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3369) 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 3369) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3369 /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(723..740) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(723..739) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(948..967) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 1067..1089 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(1127..1145) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 1213..1317 /gene="bla" /label=AmpR promoter CDS 1318..2178 /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(1536..1555) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 2349..2937 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2838..2857 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 3091..3108 /label=L4440 /note="L4440 vector, forward primer" protein_bind 3225..3246 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 3261..3291 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3299..3315 /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 3304..3326 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 3323..3339 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 3323..3339 /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 gacgcgttga cattgtagga 181 ctatattgct ctaataaagg aggcagctat gctggccgtc gttttacaac gtcgtgactg 241 ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg 301 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg 361 ctaatttact ttaaattttt tcttatgcag cctgtgatgg ataactgaat caaacaaatg 421 gcgtctgggt ttaagaagat ctgttttggc tatgttggac gaaacaagtg aacttttagg 481 atcaacttca gtttatatat ggagcttata tcgagcaata agataagtgg gctttttatg 541 taatttaatg ggctatcgtc catagattca ctaataccca tgcccagtac ccatgtatgc 601 gtttcatata agctcctaat ttctcccaca tcgctcaaat ctaaacaaat cttgttgtat 661 atataacact gagggagcaa cattggtcaa gagacctctg aagataacat actaagcttg 721 gcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat 781 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat 841 cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg gtattttctc 901 cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtac aatctgctct 961 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg 1021 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg 1081 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc 1141 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt 1201 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat 1261 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg 1321 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt 1381 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga 1441 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa 1501 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt 1561 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt 1621 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc 1681 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga 1741 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat 1801 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct 1861 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc 1921 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg 1981 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc 2041 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg 2101 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca 2161 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta 2221 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc 2281 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa 2341 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca 2401 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta 2461 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc 2521 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca 2581 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta 2641 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag 2701 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt 2761 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc 2821 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac 2881 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac 2941 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc 3001 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat 3061 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag 3121 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac 3181 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc 3241 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt 3301 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacga attcgagctc 3361 ggtacccgg //
载体图谱质粒图谱pdf版和相关资料下载
载体质粒应用举例
- 上一篇:pRGEB32
- 下一篇:pYLsgRNA-AtU3d