| 質粒名稱: | BPK3079, pBPK3079-U6-AsCpf1crRNA |
|---|---|
| 出品公司: | 愛迪基因 |
| 目錄編號: | 78741 |
| 存儲實驗室: | Keith Joung |
|
載體骨架基本信息 |
|
| 載體名稱: | MLM3636 |
| 載體出品公司: | Joung lab (Addgene Plasmid # 43860) |
| 載體抗性: | 氨芐青霉素 |
| 空載體大小: | -- |
| 完整質粒大小: | -- |
| 載體修飾: | -- |
| 載體類型: | 哺乳細胞表達,CRISPR |
| 篩選標記: | -- |
| 高拷貝/低拷貝: | 高拷貝 |
| 生長條件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名稱: | AsCpf1 crRNA backbone, without spacer sequence |
| 物種來源: | Synthetic |
| 插入大小: | -- |
| 基因突變: | -- |
| GenBank編號 | -- |
| 啟動子 | U6 |
| 標簽/融合蛋白: | -- |
| 克隆方法: | 限制性內切酶 |
| 5' 克隆位點: | BsmBI（未銷毀） |
| 3' 克隆位點: | BsmBI（未銷毀） |
| 5' 測序引物 | OS280 (5'-CAGGGTTATTGTCTCATGAGCGG-3') |
| 3' 測序引物 | -- |
|
文獻引用方法 |
|
| 材料和方法部分: | BPK3079 was a gift from Keith Joung (Addgene plasmid # 78741 ; //n2t.net/addgene:78741 ; RRID:Addgene_78741) |
| 參考文獻部分: | Genome-wide specificities of CRISPR-Cas Cpf1 nucleases in human cells. Kleinstiver BP, Tsai SQ, Prew MS, Nguyen NT, Welch MM, Lopez JM, McCaw ZR, Aryee MJ, Joung JK. Nat Biotechnol. 2016 Jun 27. doi: 10.1038/nbt.3620. 10.1038/nbt.3620 PubMed 27347757 |
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BPK3079載體 pBPK3079-U6-AsCpf1crRNA質粒基本信息
BPK3079質粒圖譜 pBPK3079-U6-AsCpf1crRNA載體圖譜和BPK3079載體序列 pBPK3079-U6-AsCpf1crRNA質粒序列多克隆位點信息
BPK3079質粒 pBPK3079-U6-AsCpf1crRNA載體簡介
BPK3079質粒序列 pBPK3079-U6-AsCpf1crRNA載體序列
LOCUS BPK3079 2225 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Human expression plasmid for AsCpf1 guide RNA (need to clone in
spacer into BsmBI sites).
ACCESSION .
VERSION .
KEYWORDS BPK3079
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 2225)
AUTHORS Kleinstiver BP, Tsai SQ, Prew MS, Nguyen NT, Welch MM, Lopez JM,
McCaw ZR, Aryee MJ, Joung JK
TITLE Genome-wide specificities of CRISPR-Cas Cpf1 nucleases in human
cells.
JOURNAL Nat Biotechnol. 2016 Jun 27. doi: 10.1038/nbt.3620.
PUBMED 27347757
REFERENCE 2 (bases 1 to 2225)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
//www.snapgene.com
FEATURES Location/Qualifiers
source 1..2225
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 82..322
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
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"
rep_origin complement(476..1064)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(556..575)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(1235..2095)
/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 1858..1877
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(2096..2200)
/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 gtaatttcta ctcttgtaga tggagacgat
361 taatgcgtct ccttttttta agcttgggcc gctcgaggta cctctctaca tatgacatgt
421 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
481 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
541 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
601 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
661 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
721 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
781 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
841 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
901 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
961 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
1021 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
1081 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
1141 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
1201 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
1261 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
1321 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
1381 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
1441 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
1501 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
1561 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
1621 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
1681 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
1741 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
1801 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
1861 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
1921 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
1981 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
2041 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
2101 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
2161 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
2221 cacct
//
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