| 质粒名称: | pYLsgRNA-AtU6-29 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 66203 |
| 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 | -- |
| 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-AtU6-29 was a gift from Yao-Guang Liu (Addgene plasmid # 66203 ; http://n2t.net/addgene:66203 ; RRID:Addgene_66203) |
| 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-AtU6-29 3162 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION cloning of sgRNAs for expression in plants.
ACCESSION .
VERSION .
KEYWORDS pYLsgRNA-AtU6-29
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3162)
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 3162)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..3162
/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(516..533)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(516..532)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(525..547)
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind complement(741..760)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 860..882
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(920..938)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 1006..1110
/gene="bla"
/label=AmpR promoter
CDS 1111..1971
/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(1329..1348)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 2142..2730
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2631..2650
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 2884..2901
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 3018..3039
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 3054..3084
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3092..3108
/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 3097..3119
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 3116..3132
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 3116..3132
/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 gaaaatatca gagatctctt
181 acagttagtt tcgttcttaa tccaaactac tgcagcctga cagacaaatg aggatgcaaa
241 caattttaaa gtttatctaa cgctagctgt tttgtttctt ctctctggtg caccaacgac
301 ggcgttttct caatcataaa gaggcttgtt ttacttaagg ccaataatgt tgatggatcg
361 aaagaagagg gcttttaata aacgagcccg tttaagctgt aaacgatgtc aaaaacatcc
421 cacatcgttc agttgaaaat agtagctctg tttatatatt ggtagagtcg actaagagat
481 tgagagacct ctgaagataa catactaagc ttggcactgg ccgtcgtttt acaacgtcgt
541 gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc
601 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
661 aatggcgaat ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
721 cgcatatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga
781 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
841 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
901 aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
961 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
1021 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
1081 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
1141 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
1201 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
1261 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
1321 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
1381 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
1441 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
1501 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
1561 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
1621 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
1681 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
1741 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
1801 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
1861 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
1921 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
1981 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
2041 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
2101 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
2161 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
2221 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
2281 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
2341 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
2401 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
2461 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
2521 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
2581 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
2641 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
2701 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
2761 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
2821 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
2881 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
2941 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
3001 agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac
3061 tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga
3121 aacagctatg accatgatta cgaattcgag ctcggtaccc gg
//
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