| 质粒名称: | pUC119-gRNA |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 52255 |
| 存储实验室: | Jen Sheen |
|
载体骨架基本信息 |
|
| 载体名称: | pUC119 |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 3162 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | CRISPR;植物表达 |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | -- |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | guide RNA targeting AtPDS3,PDS3,phytoene desaturase 3 |
| 物种来源: | A. thaliana (mustard weed) |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | KF264452 |
| 启动子 | 拟南芥U6聚合酶III启动子 |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | SacI (未破坏) |
| 3' 克隆位点: | SmaI (在克隆过程中销毁) |
| 5' 测序引物 | 5’ TGGAATTGTGAGCGGATA 3’ |
| 3' 测序引物 | 5’ ATTAAGTTGGGTAACGCC 3' |
|
文献引用方法 |
|
| 材料和方法部分: | pUC119-gRNA was a gift from Jen Sheen (Addgene plasmid # 52255 ; http://n2t.net/addgene:52255 ; RRID:Addgene_52255) |
| 参考文献部分: | Multiplex and homologous recombination-mediated genome editing in Arabidopsis and Nicotiana benthamiana using guide RNA and Cas9. Li JF, Norville JE, Aach J, McCormack M, Zhang D, Bush J, Church GM, Sheen J. Nat Biotechnol. 2013 Aug;31(8):688-91. doi: 10.1038/nbt.2654. 10.1038/nbt.2654 PubMed 23929339 |
买家导航
pUC119gRNA载体基本信息
pUC119gRNA载体图谱质粒图谱和多克隆位点信息
pUC119gRNA载体简介
pUC119gRNA载体序列
LOCUS pUC119-gRNA 3771 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Can use a PCR template to assemble new desired guide RNA. Contains
an Arabidopsis U6 promoter to drive guide RNA (targeting AtPDS3 gene
target site 1) expression with a TTTTTT as terminator. .
ACCESSION .
VERSION .
KEYWORDS pUC119-gRNA
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3771)
AUTHORS Li JF, Norville JE, Aach J, McCormack M, Zhang D, Bush J, Church GM,
Sheen J
TITLE Multiplex and homologous recombination-mediated genome editing in
Arabidopsis and Nicotiana benthamiana using guide RNA and Cas9.
JOURNAL Nat Biotechnol. 2013 Aug;31(8):688-91. doi: 10.1038/nbt.2654.
PUBMED 23929339
REFERENCE 2 (bases 1 to 3771)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..3771
/organism="synthetic DNA construct"
/mol_type="other DNA"
protein_bind 107..128
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 143..173
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 181..197
/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 186..208
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 205..221
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 205..221
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
promoter 407..710
/label=AtU6-1 promoter
/note="promoter for the Arabidopsis thaliana U6-1 snRNA
gene (Waibel and Filipowicz, 1990)"
misc_RNA 731..806
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
protein_bind complement(818..842)
/gene="mutant version of attB"
/label=attB2
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
primer_bind complement(900..917)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(900..916)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(909..931)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 1129..1584
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(1216..1235)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 1426..1447
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(1601..1620)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 1720..1742
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(1780..1798)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 1866..1970
/gene="bla"
/label=AmpR promoter
CDS 1971..2831
/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(2189..2208)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 3002..3590
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 3491..3510
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 3744..3761
/label=L4440
/note="L4440 vector, forward primer"
ORIGIN
1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgggcgc gccaagctta
241 ggcctgtcga cgcccgggcg gtaccgcgat cgctcgcgac ctgcaggcat aaagccgtca
301 gtgtccgcat aaagaaccac ccataatacc cataatagct gtttgccatc gctaccttag
361 gaccgttata gttaattacc ctgttatccc tattaattaa gagctcagaa atctcaaaat
421 tccggcagaa caattttgaa tctcgatccg tagaaacgag acggtcattg ttttagttcc
481 accacgatta tatttgaaat ttacgtgagt gtgagtgaga cttgcataag aaaataaaat
541 ctttagttgg gaaaaaattc aataatataa atgggcttga gaaggaagcg agggataggc
601 ctttttctaa aataggccca tttaagctat taacaatctt caaaagtacc acagcgctta
661 ggtaaagaaa gcagctgagt ttatatatgg ttagagacga agtagtgatt ggacttttgc
721 cagccatggt gttttagagc tagaaatagc aagttaaaat aaggctagtc cgttatcaac
781 ttgaaaaagt ggcaccgagt cggtgctttt tttctagacc cagctttctt gtacaaagtt
841 ggcattaccc atcggcgcgc ctctagaatt taaatggatc ctacgtactc gaggaattca
901 ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc
961 cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc
1021 ccttcccaac agttgcgcag cctgaatggc gaatggcgcc tgatgcggta ttttctcctt
1081 acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta
1141 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
1201 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
1261 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
1321 acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat
1381 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
1441 aaactggaac aacactcaac cctatctcgg gctattcttt tgatttataa gggattttgc
1501 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
1561 acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg
1621 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
1681 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
1741 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt
1801 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
1861 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
1921 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
1981 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
2041 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
2101 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
2161 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
2221 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
2281 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
2341 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
2401 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
2461 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
2521 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
2581 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
2641 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
2701 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
2761 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
2821 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
2881 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
2941 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
3001 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
3061 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
3121 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
3181 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
3241 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
3301 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
3361 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
3421 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
3481 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
3541 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
3601 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
3661 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
3721 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa g
//
