| 质粒名称: | BPK3082,pBPK3082-U6-LbCpf1crRNA |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 78742 |
| 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) |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达,CRISPR |
| 筛选标记: | -- |
| 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: | LbCpf1 crRNA backbone, without spacer sequence |
| 物种来源: | Synthetic |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | U6 |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | BsmBI（未销毁） |
| 3' 克隆位点: | BsmBI（未销毁） |
| 5' 测序引物 | OS280 (5'-CAGGGTTATTGTCTCATGAGCGG-3') |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | BPK3082 was a gift from Keith Joung (Addgene plasmid # 78742 ; http://n2t.net/addgene:78742 ; RRID:Addgene_78742) |
| 参考文献部分: | 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 |
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 BPK3082 2225 bp ds-DNA circular SYN 27-MAR-2019
DEFINITION Human expression plasmid for LbCpf1 guide RNA (need to clone in
spacer into BsmBI sites).
ACCESSION .
VERSION .
KEYWORDS BPK3082
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 Mar 27, 2019 from SnapGene Server 1.1.58
http://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 gaatttctac taagtgtaga 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
//
