| 质粒名称: | pFYF1548 EMX1 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 47508 |
| 存储实验室: | Keith Joung |
|
载体骨架基本信息 |
|
| 载体名称: | MLM3636 |
| 载体出品公司: | Joung lab (Addgene plasmid #43860) |
| 空载体大小: | 2278bp |
| 完整质粒大小: | 2279bp |
| 载体类型: | 哺乳细胞表达, CRISPR |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | XL1 Blue |
| 拷贝数: | 未知 |
|
插入基因信息 |
|
| 插入基因名称: | gRNA-EMX1 |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 20bp |
| 启动子: | U6 |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | BsmBI (not destroyed) |
| 3’克隆位点: | BsmBI (not destroyed) |
| 5’测序引物 | hU6-F (5'-GAGGGCCTATTTCCCATGATT-3') |
|
文献引用方法 |
|
| 材料和方法部分: |
pFYF1548 EMX1 was a gift from Keith Joung (Addgene plasmid # 47508 ; http://n2t.net/addgene:47508 ; RRID:Addgene_47508)
|
| 参考文献部分: | High-frequency off-target mutagenesis induced by CRISPR-Cas nucleases in human cells. Fu Y, Foden JA, Khayter C, Maeder ML, Reyon D, Joung JK, Sander JD. Nat Biotechnol. 2013 Jun 23. doi: 10.1038/nbt.2623. 10.1038/nbt.2623 PubMed 23792628 |
买家导航
pFYF1548 EMX1载体质粒基本信息
pFYF1548 EMX1质粒图谱载体图谱和pFYF1548 EMX1载体序列质粒序列多克隆位点信息
pFYF1548 EMX1质粒载体简介
pFYF1548 EMX1质粒序列载体序列
LOCUS pFYF1548 EMX1 2279 bp DNA circular SYN 14-APR-2025
DEFINITION human gRNA expression vector targeting EMX1.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 2279)
AUTHORS Fu Y, Foden JA, Khayter C, Maeder ML, Reyon D, Joung JK, Sander JD
TITLE High-frequency off-target mutagenesis induced by CRISPR-Cas
nucleases in human cells.
JOURNAL Nat Biotechnol. 2013 Jun 23. doi: 10.1038/nbt.2623.
PUBMED 23792628
REFERENCE 2 (bases 1 to 2279)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 14, 2025 from SnapGene Server 8.0.1
https://www.biofeng.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat
Biotechnol. 2013 Jun 23. doi: 10.1038/nbt.2623."
COMMENT Sequence Label: pFYF1548 EMX1
FEATURES Location/Qualifiers
source 1..2279
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 82..330
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA
(Domitrovich & Kunkel, 2003)"
primer_bind 82..102
/label=hU6-F
/note="Human U6 promoter, forward primer"
primer_bind 253..272
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
misc_RNA 351..426
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
rep_origin complement(530..1118)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(610..629)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(join(1289..2080,2081..2149))
/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 1912..1931
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(2150..2254)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 gacgtcgcta gctgtacaaa aaagcaggct ttaaaggaac caattcagtc gactggatcc
61 ggtaccaagg tcgggcagga agagggccta tttcccatga ttccttcata tttgcatata
121 cgatacaagg ctgttagaga gataattaga attaatttga ctgtaaacac aaagatatta
181 gtacaaaata cgtgacgtag aaagtaataa tttcttgggt agtttgcagt tttaaaatta
241 tgttttaaaa tggactatca tatgcttacc gtaacttgaa agtatttcga tttcttggct
301 ttatatatct tgtggaaagg acgaaacacc gagtccgagc agaagaagaa gttttagagc
361 tagaaatagc aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt
421 cggtgctttt tttaagcttg ggccgctcga ggtacctctc tacatatgac atgtgagcaa
481 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
541 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
601 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
661 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
721 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
781 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
841 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
901 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
961 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
1021 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
1081 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
1141 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
1201 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
1261 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
1321 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
1381 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
1441 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
1501 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
1561 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
1621 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
1681 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
1741 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
1801 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
1861 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
1921 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
1981 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
2041 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
2101 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
2161 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
2221 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacct
//
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