Basic Vector Information
- Vector Name:
- pCFD2-dU6:2gRNA
- Antibiotic Resistance:
- Ampicillin
- Length:
- 6227 bp
- Type:
- CRISPR Plasmids
- Replication origin:
- ori
- Source/Author:
- Port F, Chen HM, Lee T, Bullock SL.
- Copy Number:
- High copy number
- Promoter:
- dU6-2
pCFD2-dU6:2gRNA vector Map
pCFD2-dU6:2gRNA vector Sequence
LOCUS pCFD2-dU6-2gRNA. 6227 bp DNA circular SYN 01-JAN-1980 DEFINITION Drosophila expression vector in which a guide RNA (gRNA) is expressed from the U6-2 promoter. ACCESSION . VERSION . KEYWORDS pCFD2-dU6-2gRNA SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6227) AUTHORS Port F, Chen HM, Lee T, Bullock SL. TITLE Optimized CRISPR/Cas tools for efficient germline and somatic genome engineering in Drosophila. JOURNAL Proc. Natl. Acad. Sci. U.S.A. 2014;111:E2967-76. PUBMED 25002478 REFERENCE 2 (bases 1 to 6227) AUTHORS Bullock Lab / Addgene #49409 TITLE Direct Submission REFERENCE 3 (bases 1 to 6227) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Proc. Natl. Acad. Sci. U.S.A."; date: "2014"; volume: "111"; pages: "E2967-76" COMMENT SGRef: number: 2; type: "Journal Article" FEATURES Location/Qualifiers source 1..6227 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 7..406 /label=dU6-2 promoter /note="RNA polymerase III promoter for Drosophila U6-2 snRNA (Das et al., 1987)" misc_RNA 425..500 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" terminator 501..506 /note="polIII terminator" /note="RNA polymerase III transcription terminator" promoter complement(811..829) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(850..866) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind complement(874..890) /label=lac operator /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(898..928) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind complement(943..964) /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(1252..1840) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(2014..2871) /label=AmpR /note="beta-lactamase" promoter complement(2872..2976) /label=AmpR promoter rep_origin complement(3002..3457) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind 3599..3615 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" promoter 3622..3640 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS complement(join(3832..3949,4026..4119,4211..4817,4872..5005, 5066..5226,5300..5325)) /codon_start=1 /product="tryptophan oxygenase" /label=tryptophan oxygenase /note="vermilion" /note="selectable marker, required to synthesize the brown eye pigment in Drosophila" /translation="MSCPYAGNGNDHDDSAVPLTTEVGKIYGEYLMLDKLLDAQCMLSE EDKRPVHDEHLFIITHQAYELWFKQIIFEFDSIRDMLDAEVIDETKTLEIVKRLNRVVL ILKLLVDQVPILETMTPLDFMDFRKYLAPASGFQSLQFRLIENKLGVLTEQRVRYNQKY SDVFSDEEARNSIRNSEKDPSLLELVQRWLERTPGLEESGFNFWAKFQESVDRFLEAQV QSAMEEPVEKAKNYRLMDIEKRREVYRSIFDPAVHDALVRRGDRRFSHRALQGAIMITF YRDEPRFSQPHQLLTLLMDIDSLITKWRYNHVIMVQRMIGSQQLGTGGSSGYQYLRSTL SDRYKVFLDLFNLSTFLIPREAIPPLDETIRKKLINKSV" protein_bind 5640..5709 /label=attB /note="attB site for the phi-C31 integrase (Groth et al., 2000)"
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