载体基本信息 | |||
载体名称 | CRISPRi library backbone | 载体抗性 | Ampicillin |
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载体长度 | 8888 bp | 载体类型 | CRISPR Plasmids |
载体来源 | Gilbert LA, Horlbeck MA, Adamson B, Villalta JE, Chen Y, Whitehead | 拷贝数 | High copy number |
CRISPRi library backbone 载体载体图谱
CRISPRi library backbone 载体载体序列
LOCUS CRISPRi_library 8888 bp DNA circular SYN 01-JAN-1980 DEFINITION Backbone vector for generating a CRISPR-mediated transcriptional repression (CRISPRi) library. ACCESSION . VERSION . KEYWORDS CRISPRi library backbone. SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8888) AUTHORS Gilbert LA, Horlbeck MA, Adamson B, Villalta JE, Chen Y, Whitehead EH, Guimaraes C, Panning B, Ploegh HL, Bassik MC, Qi LS, Kampmann M, Weissman JS. TITLE Genome-Scale CRISPR-Mediated Control of Gene Repression and Activation. JOURNAL Cell 2014;159:647-61. PUBMED 25307932 REFERENCE 2 (bases 1 to 8888) AUTHORS Weissman Lab / Addgene #62217 TITLE Direct Submission REFERENCE 3 (bases 1 to 8888) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Cell"; date: "2014"; volume: "159"; pages: "647-61" COMMENT SGRef: number: 2; type: "Journal Article" FEATURES Location/Qualifiers source 1..8888 /mol_type="other DNA" /organism="synthetic DNA construct" enhancer 247..626 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 627..829 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" LTR 844..1024 /label=LTR (truncated) /note="LTR (truncated)" /note="truncated long terminal repeat (LTR) from HIV-1" misc_feature 1071..1196 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1695..1928 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 2113..2157 /label=gp41 peptide /note="antigenic peptide corresponding to amino acids 655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik et al., 2013)" CDS 2306..2347 /note="Protein Tat from Human immunodeficiency virus type 1 group M subtype B (isolate WMJ22). Accession#: P12509" misc_feature 2455..2572 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 2631..2943 /note="U6 promoter" /note="modified RNA polymerase III promoter for mouse U6 snRNA" protein_bind 2825..2858 /label=Cre recombinase binding site /bound_moiety="Cre recombinase" /note="loxP" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." misc_feature 2965..3050 /label=gRNA scaffold /note="gRNA scaffold" /note="modified guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" terminator 3051..3056 /note="polIII terminator" /note="RNA polymerase III transcription terminator" promoter 3114..4286 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" CDS 4298..4894 /label=PuroR /note="puromycin N-acetyltransferase" CDS 4904..4957 /codon_start=1 /product="2A peptide from Thosea asigna virus capsid protein" /label=2A peptide from Thosea asigna virus capsid protein /note="T2A" /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /translation="EGRGSLLTCGDVEENPGP" CDS 4970..5665 /label=TagBFP /note="monomeric blue fluorescent protein" protein_bind complement(5688..5721) /label=loxP /note="Cre-mediated recombination occurs in the 8-bp core sequence (ATGTATGC) (Shaw et al., 2021)." misc_feature 5777..6365 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(6368..6384) /label=KS primer /note="common sequencing primer, one of multiple similar variants" LTR 6894..7074 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" rep_origin complement(7136..7724) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(7898..8755) /label=AmpR /note="beta-lactamase" promoter complement(8756..8860) /label=AmpR promoter
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