| 质粒名称: | pYLsgRNA-AtU6-1 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 66202 |
| 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-AtU6-1 was a gift from Yao-Guang Liu (Addgene plasmid # 66202 ; http://n2t.net/addgene:66202 ; RRID:Addgene_66202) |
| 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-AtU6-1 3146 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION cloning of sgRNAs for expression in plants.
ACCESSION .
VERSION .
KEYWORDS pYLsgRNA-AtU6-1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3146)
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 3146)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..3146
/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..465
/label=AtU6-1 promoter
/note="promoter for the Arabidopsis thaliana U6-1 snRNA
gene (Waibel and Filipowicz, 1990)"
primer_bind complement(500..517)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(500..516)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(509..531)
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind complement(725..744)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 844..866
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(904..922)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 990..1094
/gene="bla"
/label=AmpR promoter
CDS 1095..1955
/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(1313..1332)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 2126..2714
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2615..2634
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 2868..2885
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 3002..3023
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 3038..3068
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3076..3092
/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 3081..3103
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 3100..3116
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 3100..3116
/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 gagaaatctc aaaattccgg
181 cagaacaatt ttgaatctcg atccgtagaa acgagacggt cattgtttta gttccaccac
241 gattatattt gaaatttacg tgagtgtgag tgagacttgc ataagaaaat aaaatcttta
301 gttgggaaaa aattcaataa tataaatggg cttgagaagg aagcgaggga taggcctttt
361 tctaaaatag gcccatttaa gctattaaca atcttcaaaa gtaccacagc gcttaggtaa
421 agaaagcagc tgagtttata tatggttaga gacgaagtag tgattgagag acctctgaag
481 ataacatact aagcttggca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg
541 gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg
601 aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggcgcc
661 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata tggtgcactc
721 tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg
781 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
841 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa
901 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
961 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
1021 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
1081 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
1141 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
1201 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
1261 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
1321 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
1381 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
1441 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
1501 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
1561 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
1621 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
1681 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
1741 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
1801 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
1861 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
1921 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
1981 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
2041 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
2101 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
2161 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
2221 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
2281 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
2341 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
2401 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
2461 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
2521 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
2581 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
2641 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
2701 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
2761 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
2821 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
2881 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
2941 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
3001 ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc
3061 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg
3121 attacgaatt cgagctcggt acccgg
//
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