| 载体基本信息 |
| 载体名称 |
BPK3079 |
载体抗性 |
Ampicillin |
| 载体长度 |
2225 bp |
载体类型 |
Mammalian Expression, CRISPR |
| 拷贝数 |
High Copy |
启动子 |
U6 |
| 克隆方法 |
Restriction Enzyme |
5引物 |
OS280 (5-CAGGGTTATTGTCTCATGAGCGG-3) |
BPK3079 载体载体图谱
BPK3079 载体载体序列
LOCUS Exported 2225 bp ds-DNA circular SYN 13-MAY-2021
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
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat
Biotechnol. 2016 Jun 27. doi: 10.1038/nbt.3620."
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