| 质粒名称: | pCRISPRainbow-DONOR1 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 75398 |
| 存储实验室: | Thoru Pederson |
|
载体骨架基本信息 |
|
| 载体名称: | pUC19 |
| 载体抗性: | 氯霉素和氨苄青霉素 |
| 空载体大小: | 2000 bp |
| 完整质粒大小: | 3524 bp |
| 载体类型: | 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: | Cm(R)-CcdB |
| 启动子 | Bacteria |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | Mlu I(未破坏) |
| 3' 克隆位点: | Pci I (未破坏) |
| 5' 测序引物 | GCTACAACAAGGCAAGGCTTGACCGA |
| 3' 测序引物 | AACGCCAGCAACGCGGCCTT |
|
文献引用方法 |
|
| 材料和方法部分: | pCRISPRainbow-DONOR1 was a gift from Thoru Pederson (Addgene plasmid # 75398 ; http://n2t.net/addgene:75398 ; RRID:Addgene_75398) |
| 参考文献部分: | Multiplexed labeling of genomic loci with dCas9 and engineered sgRNAs using CRISPRainbow. Ma H, Tu LC, Naseri A, Huisman M, Zhang S, Grunwald D, Pederson T. Nat Biotechnol. 2016 Apr 18. doi: 10.1038/nbt.3526. 10.1038/nbt.3526 PubMed 27088723 |
买家导航
pCRISPRainbowDONOR1载体基本信息
pCRISPRainbowDONOR1载体图谱质粒图谱和多克隆位点信息
pCRISPRainbowDONOR1载体简介
pCRISPRainbowDONOR1载体序列
LOCUS pCRISPRainbow-DONOR1 3527 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION CRISPRainbow-DONOR1.
ACCESSION .
VERSION .
KEYWORDS pCRISPRainbow-DONOR1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3527)
AUTHORS Ma H, Tu LC, Naseri A, Huisman M, Zhang S, Grunwald D, Pederson T
TITLE Multiplexed labeling of genomic loci with dCas9 and engineered
sgRNAs using CRISPRainbow.
JOURNAL Nat Biotechnol. 2016 Apr 18. doi: 10.1038/nbt.3526.
PUBMED 27088723
REFERENCE 2 (bases 1 to 3527)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..3527
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 185..215
/label=lac UV5 promoter
/note="E. coli lac promoter with an ""up"" mutation"
CDS 269..928
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
primer_bind complement(335..354)
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
CDS 1270..1575
/codon_start=1
/gene="ccdB"
/product="CcdB, a bacterial toxin that poisons DNA gyrase"
/label=ccdB
/note="Plasmids containing the ccdB gene cannot be
propagated in standard E. coli strains."
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
primer_bind 1454..1473
/label=ccdB-fwd
/note="ccdB gene, forward primer"
rep_origin complement(1688..2276)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(1768..1787)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(2447..3307)
/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 3070..3089
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(3308..3412)
/gene="bla"
/label=AmpR promoter
primer_bind complement(3487..3506)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
ORIGIN
1 tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa
61 ggcaaggctt gaccgacaat tccatgaaga atctgcttag ggttaggcgt tttgcgctgc
121 ttcgcgatgt acgggccaga tatacgcgtt gacagagacc gcggccgcat taggcacccc
181 aggctttaca ctttatgctt ccggctcgta taatgtgtgg attttgagtt aggatccgtc
241 gagattttca ggagctaagg aagctaaaat ggagaaaaaa atcactggat ataccaccgt
301 tgatatatcc caatggcatc gtaaagaaca ttttgaggca tttcagtcag ttgctcaatg
361 tacctataac cagaccgttc agctggatat tacggccttt ttaaagaccg taaagaaaaa
421 taagcacaag ttttatccgg cctttattca cattcttgcc cgcctgatga atgctcatcc
481 ggaattccgt atggcaatga aagacggtga gctggtgata tgggatagtg ttcacccttg
541 ttacaccgtt ttccatgagc aaactgaaac gttttcatcg ctctggagtg aataccacga
601 cgatttccgg cagtttctac acatatattc gcaagatgtg gcgtgttacg gtgaaaacct
661 ggcctatttc cctaaagggt ttattgagaa tatgtttttc gtctcagcca atccctgggt
721 gagtttcacc agttttgatt taaacgtggc caatatggac aacttcttcg cccccgtttt
781 caccatgggc aaatattata cgcaaggcga caaggtgctg atgccgctgg cgattcaggt
841 tcatcatgcc gtttgtgatg gcttccatgt cggcagaatg cttaatgaat tacaacagta
901 ctgcgatgag tggcagggcg gggcgtaaag atctggatcc ggcttactaa aagccagata
961 acagtatgcg tatttgcgcg ctgatttttg cggtataaga atatatactg atatgtatac
1021 ccgaagtatg tcaaaaagag gtatgctatg aagcagcgta ttacagtgac agttgacagc
1081 gacagctatc agttgctcaa ggcatatatg atgtcaatat ctccggtctg gtaagcacaa
1141 ccatgcagaa tgaagcccgt cgtctgcgtg ccgaacgctg gaaagcggaa aatcaggaag
1201 ggatggctga ggtcgcccgg tttattgaaa tgaacggctc ttttgctgac gagaacaggg
1261 gctggtgaaa tgcagtttaa ggtttacacc tataaaagag agagccgtta tcgtctgttt
1321 gtggatgtac agagtgatat tattgacacg cccgggcgac ggatggtgat ccccctggcc
1381 agtgcacgtc tgctgtcaga taaagtctcc cgtgaacttt acccggtggt gcatatcggg
1441 gatgaaagct ggcgcatgat gaccaccgat atggccagtg tgccggtctc cgttatcggg
1501 gaagaagtgg ctgatctcag ccaccgcgaa aatgacatca aaaacgccat taacctgatg
1561 ttctggggaa tataaatgtc aggctccctt atacacagcc agtctgcagg tcgacggtct
1621 cgactcacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
1681 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
1741 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
1801 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
1861 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
1921 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
1981 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
2041 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
2101 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
2161 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
2221 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
2281 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
2341 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
2401 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
2461 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
2521 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
2581 taccgcgaga accacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
2641 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
2701 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
2761 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
2821 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
2881 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
2941 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
3001 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
3061 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
3121 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
3181 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
3241 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
3301 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
3361 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
3421 cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtgcac
3481 tctcagtaca atctgctctg atgccgcata gttaagccag tatctgc
//
pCRISPRainbowDONOR1载体图谱质粒图谱pdf版和相关资料下载
pCRISPRainbowDONOR1载体质粒应用举例
- 上一篇：pcDNA-dCas9
- 下一篇：pLH-sgRNA1
