| 質粒名稱: | pXR003: CasRx gRNA cloning backbone |
|---|---|
| 出品公司: | 愛迪基因 |
| 目錄編號: | 109053 |
| 存儲實驗室: | Patrick Hsu |
|
載體骨架基本信息 |
|
| 載體名稱: | pUC19 |
| 載體出品公司: | -- |
| 載體抗性: | 氨芐青霉素 |
| 空載體大小: | 2686 bp |
| 完整質粒大小: | -- |
| 載體修飾: | inserted gRNA expression cassette |
| 載體類型: | 哺乳動物表達,CRISPR |
| 篩選標記: | -- |
| 高拷貝/低拷貝: | 高拷貝 |
| 生長條件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名稱: | -- |
| 物種來源: | -- |
| 插入大小: | -- |
| 基因突變: | -- |
| GenBank編號 | -- |
| 啟動子 | hU6 |
| 標簽/融合蛋白: | -- |
| 克隆方法: | -- |
| 5' 克隆位點: | -- |
| 3' 克隆位點: | -- |
| 5' 測序引物 | -- |
| 3' 測序引物 | -- |
|
文獻引用方法 |
|
| 材料和方法部分: | pXR003: CasRx gRNA cloning backbone was a gift from Patrick Hsu (Addgene plasmid # 109053 ; //n2t.net/addgene:109053 ; RRID:Addgene_109053) |
| 參考文獻部分: | Transcriptome Engineering with RNA-Targeting Type VI-D CRISPR Effectors. Konermann S, Lotfy P, Brideau NJ, Oki J, Shokhirev MN, Hsu PD. Cell. 2018 Mar 8. pii: S0092-8674(18)30207-1. doi: 10.1016/j.cell.2018.02.033. 10.1016/j.cell.2018.02.033 PubMed 29551272 |
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pXR003: CasRx gRNA cloning backbone載體質粒基本信息
pXR003: CasRx gRNA cloning backbone質粒圖譜載體圖譜和pXR003: CasRx gRNA cloning backbone載體序列質粒序列多克隆位點信息
pXR003: CasRx gRNA cloning backbone質粒載體簡介
pXR003: CasRx gRNA cloning backbone質粒序列載體序列
LOCUS pXR003: CasRx gRNA cloning backbone 2903 bp ds-DNA circular SYN 24-JUN-2019
DEFINITION hU6-driven expression of guide RNAs compatible with CasRx. 5'
processed DR followed by BbsI sites for guide cloning..
ACCESSION .
VERSION .
KEYWORDS pXR003: CasRx gRNA cloning backbone
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 2903)
AUTHORS Konermann S, Lotfy P, Brideau NJ, Oki J, Shokhirev MN, Hsu PD
TITLE Transcriptome Engineering with RNA-Targeting Type VI-D CRISPR
Effectors.
JOURNAL Cell. 2018 Mar 8. pii: S0092-8674(18)30207-1. doi:
10.1016/j.cell.2018.02.033.
PUBMED 29551272
REFERENCE 2 (bases 1 to 2903)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Jun 24, 2019 from SnapGene Server 1.1.58
//www.snapgene.com
FEATURES Location/Qualifiers
source 1..2903
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 7..247
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
primer_bind 7..27
/label=hU6-F
/note="Human U6 promoter, forward primer"
primer_bind 178..197
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
rep_origin 386..841
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(473..492)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 683..704
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(858..877)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 977..999
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(1037..1055)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 1123..1227
/gene="bla"
/label=AmpR promoter
CDS 1228..2088
/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(1446..1465)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 2259..2847
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2748..2767
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 gctagcgagg gcctatttcc catgattcct tcatatttgc atatacgata caaggctgtt
61 agagagataa ttggaattaa tttgactgta aacacaaaga tattagtaca aaatacgtga
121 cgtagaaagt aataatttct tgggtagttt gcagttttaa aattatgttt taaaatggac
181 tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct tggctttata tatcttgtgg
241 aaaggacgaa acaccgaacc cctaccaact ggtcggggtt tgaaacgggt cttcgagaag
301 accttttttt tgaattctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
361 ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt
421 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc cgctcctttc
481 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
541 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
601 ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
661 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaactct
721 atctcgggct attcttttga tttataaggg attttgccga tttcggtcta ttggttaaaa
781 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt
841 ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac
901 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
961 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
1021 cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
1081 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
1141 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
1201 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
1261 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
1321 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
1381 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
1441 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
1501 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
1561 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
1621 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
1681 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
1741 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
1801 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
1861 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
1921 aaatctggag ccggtgagcg tggaagccgc ggtatcattg cagcactggg gccagatggt
1981 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
2041 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
2101 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
2161 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
2221 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
2281 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
2341 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
2401 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
2461 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
2521 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
2581 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
2641 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
2701 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
2761 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
2821 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
2881 gccttttgct ggccttttgc tca
//