| 質粒名稱: | sgRNA(MS2) cloning backbone |
|---|---|
| 出品公司: | 愛迪基因 |
| 目錄編號: | 61424 |
| 存儲實驗室: | Feng Zhang |
|
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: | -- |
| 完整質粒大小: | -- |
| 載體修飾: | -- |
| 載體類型: | 哺乳細胞表達 |
| 篩選標記: | 新霉素（G418） |
| 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 | -- |
| 標簽/融合蛋白: | -- |
| 克隆方法: | 限制性內切酶 |
| 5' 克隆位點: | AgeI(未破壞) |
| 3' 克隆位點: | NotI (未破壞) |
| 5' 測序引物 | CMV-F |
| 3' 測序引物 | SV40pA-R |
|
文獻引用方法 |
|
| 材料和方法部分: | sgRNA(MS2) cloning backbone was a gift from Feng Zhang (Addgene plasmid # 61424 ; //n2t.net/addgene:61424 ; RRID:Addgene_61424) |
| 參考文獻部分: | Genome-scale transcriptional activation by an engineered CRISPR-Cas9 complex. Konermann S, Brigham MD, Trevino AE, Joung J, Abudayyeh OO, Barcena C, Hsu PD, Habib N, Gootenberg JS, Nishimasu H, Nureki O, Zhang F. Nature. 2014 Dec 10. doi: 10.1038/nature14136. 10.1038/nature14136 PubMed 25494202 |
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sgRNA(MS2) cloning backbone載體質粒基本信息
sgRNA(MS2) cloning backbone質粒圖譜載體圖譜和sgRNA(MS2) cloning backbone載體序列質粒序列多克隆位點信息
 cloning backbone質粒圖譜.png)
sgRNA(MS2) cloning backbone質粒載體簡介
sgRNA(MS2) cloning backbone質粒序列載體序列
LOCUS sgRNA(MS2) cloning backbone 3008 bp ds-DNA circular SYN 12-AUG-2019
DEFINITION sgRNA cloning backbone with MS2 loops at tetraloop and stemloop 2.
Contains BbsI sites for insertion of spacer sequences..
ACCESSION .
VERSION .
KEYWORDS sgRNA(MS2) cloning backbone
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3008)
AUTHORS Konermann S, Brigham MD, Trevino AE, Joung J, Abudayyeh OO, Barcena
C, Hsu PD, Habib N, Gootenberg JS, Nishimasu H, Nureki O, Zhang F
TITLE Genome-scale transcriptional activation by an engineered CRISPR-Cas9
complex.
JOURNAL Nature. 2014 Dec 10. doi: 10.1038/nature14136.
PUBMED 25494202
REFERENCE 2 (bases 1 to 3008)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 12, 2019 from SnapGene Server 1.1.58
//www.snapgene.com
FEATURES Location/Qualifiers
source 1..3008
/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"
misc_RNA 291..309
/label=MS2 stem loop
/note="stem loop that binds the bacteriophage MS2 coat
protein"
misc_RNA 361..379
/label=MS2 stem loop
/note="stem loop that binds the bacteriophage MS2 coat
protein"
rep_origin 491..946
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(578..597)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 788..809
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(963..982)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 1082..1104
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(1142..1160)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 1228..1332
/gene="bla"
/label=AmpR promoter
CDS 1333..2193
/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(1551..1570)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 2364..2952
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2853..2872
/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 acaccgggtc ttcgagaaga cctgttttag agctaggcca acatgaggat
301 cacccatgtc tgcagggcct agcaagttaa aataaggcta gtccgttatc aacttggcca
361 acatgaggat cacccatgtc tgcagggcca agtggcaccg agtcggtgct ttttttgaat
421 tctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag
481 caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
541 agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc
601 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
661 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca
721 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
781 tttaatagtg gactcttgtt ccaaactgga acaacactca actctatctc gggctattct
841 tttgatttat aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa
901 caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc
961 agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct
1021 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
1081 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag
1141 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
1201 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
1261 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
1321 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
1381 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
1441 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
1501 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
1561 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
1621 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
1681 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
1741 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
1801 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
1861 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
1921 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
1981 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
2041 gagcgtggaa gccgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
2101 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
2161 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
2221 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
2281 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
2341 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
2401 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
2461 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
2521 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
2581 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
2641 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
2701 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
2761 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
2821 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
2881 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
2941 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
3001 tttgctca
//
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