BPK3079 载体 (V007404#)

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

BPK30792225 bp60012001800U6 promoteroriAmpRAmpR promoter

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

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