| 质粒名称: | pC0040-LwaCas13a crRNA backbone |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 103851 |
| 存储实验室: | Feng Zhang |
|
载体骨架基本信息 |
|
| 载体名称: | pUC19 |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 2686 bp |
| 完整质粒大小: | 2962 bp |
| 载体类型: | 哺乳细胞表达,CRISPR |
| 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: | U6-LwaCas13aDR-BbsI-BbsI-polyT |
| 插入大小: | 314 bp |
| 启动子 | U6 |
| 克隆方法: | Gibson克隆 |
| 5' 测序引物 | GAGGGCCTATTTCCCATGATTCC |
|
文献引用方法 |
|
| 材料和方法部分: | pC0040-LwaCas13a crRNA backbone was a gift from Feng Zhang (Addgene plasmid # 103851 ; http://n2t.net/addgene:103851 ; RRID:Addgene_103851) |
| 参考文献部分: | RNA editing with CRISPR-Cas13. Cox DBT, Gootenberg JS, Abudayyeh OO, Franklin B, Kellner MJ, Joung J, Zhang F. Science. 2017 Oct 25. pii: eaaq0180. doi: 10.1126/science.aaq0180. 10.1126/science.aaq0180 PubMed 29070703 |
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 pC0040-LwaCas13a crRNA backbone 2962 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Backbone for cloning guides that are compatible with LwaCas13a.
Contains a 5' direct repeat. Clone using BbsI (BpiI). F overhang
AAAC. R overhang AAAA.
ACCESSION .
VERSION .
KEYWORDS pC0040-LwaCas13a crRNA backbone
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 2962)
AUTHORS Cox DBT, Gootenberg JS, Abudayyeh OO, Franklin B, Kellner MJ, Joung
J, Zhang F
TITLE RNA editing with CRISPR-Cas13.
JOURNAL Science. 2017 Oct 25. pii: eaaq0180. doi: 10.1126/science.aaq0180.
PUBMED 29070703
REFERENCE 2 (bases 1 to 2962)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..2962
/organism="synthetic DNA construct"
/mol_type="other DNA"
rep_origin complement(38..626)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(118..137)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(797..1657)
/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 1420..1439
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(1658..1762)
/gene="bla"
/label=AmpR promoter
primer_bind 1830..1848
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(1886..1908)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 2008..2027
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 2221..2243
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 2235..2252
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 2236..2252
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter complement(2344..2584)
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
primer_bind complement(2394..2413)
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
primer_bind complement(2564..2584)
/label=hU6-F
/note="Human U6 promoter, forward primer"
primer_bind complement(2598..2614)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2598..2614)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(2611..2633)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 2622..2638
/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)."
promoter complement(2646..2676)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2691..2712
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(2829..2846)
/label=L4440
/note="L4440 vector, forward primer"
ORIGIN
1 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
61 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
121 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
181 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
241 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
301 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
361 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
421 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
481 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
541 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
601 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
661 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
721 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
781 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
841 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
901 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
961 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
1021 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
1081 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
1141 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
1201 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
1261 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
1321 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
1381 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
1441 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
1501 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
1561 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
1621 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
1681 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
1741 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
1801 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg
1861 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
1921 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
1981 gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc
2041 atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt
2101 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
2161 gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt
2221 cccagtcacg acgttgtaaa acgacggcca gtgaattcga gctcggtacc aaaaaattgt
2281 cttcgtccca ggaagacatg ttttagtccc cttcgttttt ggggtagtct aaatcggtgt
2341 ttcgtccttt ccacaagata tataaagcca agaaatcgaa atactttcaa gttacggtaa
2401 gcatatgata gtccatttta aaacataatt ttaaaactgc aaactaccca agaaattatt
2461 actttctacg tcacgtattt tgtactaata tctttgtgtt tacagtcaaa ttaattccaa
2521 ttatctctct aacagccttg tatcgtatat gcaaatatga aggaatcatg ggaaataggc
2581 cctctggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
2641 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
2701 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
2761 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
2821 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
2881 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
2941 atgtgagcaa aaggccagca aa
//
