| 质粒名称: | sgRNA(MS2) cloning backbone |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 61424 |
| 存储实验室: | Feng Zhang |
|
载体骨架基本信息 |
|
| 载体名称: | pUC19 |
| 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: | -- |
| 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: | -- |
| 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: | sgRNA(MS2) cloning backbone was a gift from Feng Zhang (Addgene plasmid # 61424 ; http://n2t.net/addgene:61424 ; RRID:Addgene_61424) |
| 参考文献部分: | 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 |
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
 cloning backbone质粒图谱.png)
载体简介
载体序列
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
http://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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