| 质粒名称: | pC016-LwCas13a guide expression backbone with U6 promoter |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 91906 |
| 存储实验室: | Vladislav Verkhusha |
|
载体骨架基本信息 |
|
| 载体名称: | pUC19 |
| 载体抗性: | 氨苄霉素 |
| 空载体大小: | 2850 bp |
| 完整质粒大小: | 2908 bp |
| 载体类型: | 哺乳细胞表达 |
| 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: | LwCas13a Guide site |
| 物种来源: | 合成 |
| 插入大小: | 58 bp |
|
文献引用方法 |
|
| 材料和方法部分: | pC016 - LwCas13a guide expression backbone with U6 promoter was a gift from Feng Zhang (Addgene plasmid # 91906 ; http://n2t.net/addgene:91906 ; RRID:Addgene_91906) |
| 参考文献部分: |
RNA targeting with CRISPR-Cas13. Abudayyeh OO, Gootenberg JS, Essletzbichler P, Han S, Joung J, Belanto JJ, Verdine V, Cox DBT, Kellner MJ, Regev A, Lander ES, Voytas DF, Ting AY, Zhang F. Nature. 2017 Oct 4. doi: 10.1038/nature24049. 10.1038/nature24049 PubMed 28976959
|
pC016-LwCas13a guide expression backbone with U6 promoter
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 pC016-LwCas13a guide expression backbone with U6 promoter 2908 bp ds-DNA circular SYN 21-JUL-2020
DEFINITION Backbone for cloning LwCas13a guides under U6 promoter.
ACCESSION .
VERSION .
KEYWORDS pC016 - LwCas13a guide expression backbone with U6 promoter
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 2908)
AUTHORS Abudayyeh OO, Gootenberg JS, Essletzbichler P, Han S, Joung J,
Belanto JJ, Verdine V, Cox DBT, Kellner MJ, Regev A, Lander ES,
Voytas DF, Ting AY, Zhang F
TITLE RNA targeting with CRISPR-Cas13.
JOURNAL Nature. 2017 Oct 4. doi: 10.1038/nature24049.
PUBMED 28976959
REFERENCE 2 (bases 1 to 2908)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Jul 21, 2020 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..2908
/organism="synthetic DNA construct"
/mol_type="other DNA"
rep_origin 138..726
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 627..646
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
promoter 789..1029
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
primer_bind 789..809
/label=hU6-F
/note="Human U6 promoter, forward primer"
primer_bind 960..979
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
rep_origin 1173..1628
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(1260..1279)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 1470..1491
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(1645..1664)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 1764..1786
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(1824..1842)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 1910..2014
/gene="bla"
/label=AmpR promoter
CDS 2015..2875
/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(2233..2252)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
ORIGIN
1 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
61 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
121 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
181 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
241 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
301 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
361 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
421 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
481 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
541 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
601 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
661 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
721 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
781 cagctagcga gggcctattt cccatgattc cttcatattt gcatatacga tacaaggctg
841 ttagagagat aattggaatt aatttgactg taaacacaaa gatattagta caaaatacgt
901 gacgtagaaa gtaataattt cttgggtagt ttgcagtttt aaaattatgt tttaaaatgg
961 actatcatat gcttaccgta acttgaaagt atttcgattt cttggcttta tatatcttgt
1021 ggaaaggacg aaacaccgat ttagactacc ccaaaaacga aggggactaa aacgggtctt
1081 cgagaagacc ttttttttga attctgatgc ggtattttct ccttacgcat ctgtgcggta
1141 tttcacaccg catacgtcaa agcaaccata gtacgcgccc tgtagcggcg cattaagcgc
1201 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgcct tagcgcccgc
1261 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
1321 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
1381 acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
1441 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
1501 caactctatc tcgggctatt cttttgattt ataagggatt ttgccgattt cggtctattg
1561 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt
1621 tacaatttta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc
1681 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
1741 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
1801 accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
1861 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
1921 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
1981 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
2041 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
2101 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
2161 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
2221 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
2281 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
2341 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
2401 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
2461 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
2521 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
2581 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
2641 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
2701 tgctgataaa tctggagccg gtgagcgtgg aagccgcggt atcattgcag cactggggcc
2761 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
2821 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
2881 agaccaagtt tactcatata tactttag
//
