Basic Vector Information
- Vector Name:
- pUASTattB-LT3-Dam
- Antibiotic Resistance:
- Ampicillin
- Length:
- 10106 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Southall TD, Gold KS, Egger B, Davidson CM, Caygill EE, Marshall OJ, Brand AH.
- Promoter:
- hsp70
pUASTattB-LT3-Dam vector Vector Map
Plasmid Resuspension Protocol:
1. Centrifuge at 5,000×g for 5 min.
2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.
3. Close the tube and incubate for 10 minutes at room temperature.
4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.
5.Store the plasmid at -20 ℃.
pUASTattB-LT3-Dam vector Sequence
LOCUS 40924_44769 10106 bp DNA circular SYN 18-DEC-2018 DEFINITION Cloning vector pUASTattB-LT3-Dam, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10106) AUTHORS Southall TD, Gold KS, Egger B, Davidson CM, Caygill EE, Marshall OJ, Brand AH. TITLE Cell-type-specific profiling of gene expression and chromatin binding without cell isolation: assaying RNA Pol II occupancy in neural stem cells JOURNAL Dev. Cell 26 (1), 101-112 (2013) PUBMED 23792147 REFERENCE 2 (bases 1 to 10106) AUTHORS Southall TD, Brand AH. TITLE Direct Submission JOURNAL Submitted (18-FEB-2016) Life Sciences, Imperial College London, Sir Ernst Chain Building, London SW7 2AZ, United Kingdom REFERENCE 3 (bases 1 to 10106) TITLE Direct Submission REFERENCE 4 (bases 1 to 10106) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Dev. Cell"; date: "2013"; volume: "26"; issue: "1"; pages: "101-112" COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted (18-FEB-2016) Life Sciences, Imperial College London, Sir Ernst Chain Building, London SW7 2AZ, United Kingdom" COMMENT SGRef: number: 3; type: "Journal Article" COMMENT ##Assembly-Data-START## Sequencing Technology :: Sanger dideoxy sequencing ##Assembly-Data-END## FEATURES Location/Qualifiers source 1..10106 /mol_type="other DNA" /organism="synthetic DNA construct" rep_origin 4..459 /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind 600..616 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" promoter 623..641 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" gene 666..4802 /label=mini-white /note="This modified version of the white gene lacks part of the first intron." protein_bind 4809..4842 /label=loxP /note="Cre-mediated recombination occurs in the 8-bp core sequence (ATGTATGC) (Shaw et al., 2021)." protein_bind 4875..4969 /label=5X UAS /note="five tandem copies of the 'ScaI site' 17-mer CGGAGTACTGTCCTCCG, an upstream activating sequence (UAS) that efficiently binds yeast Gal4 (Webster et al., 1988; Pfeiffer et al., 2010)" promoter 4988..5226 /label=hsp70 promoter /note="Drosophila melanogaster hsp70Bb promoter" CDS 5261..5968 /label=mCherry /note="monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)" CDS 5991..6824 /gene="dam" /label=dam /note="DNA adenine methylase from Escherichia coli (strain K12). Accession#: P0AEE8" CDS 6831..6860 /label=Myc /note="Myc (human c-Myc proto-oncogene) epitope tag" intron 6980..7045 /label=small t intron /note="SV40 (simian virus 40) small t antigen intron" CDS 7175..7195 /label=SV40 NLS /note="nuclear localization signal of SV40 (simian virus 40) large T antigen" polyA_signal 7467..7601 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" protein_bind 7630..7699 /label=attB /note="attB site for the phi-C31 integrase (Groth et al., 2000)" promoter complement(7918..7936) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(7957..7973) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind 7981..7997 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /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(8005..8035) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind complement(8050..8071) /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(8359..8947) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(9121..9978) /label=AmpR /note="beta-lactamase" promoter complement(9979..10083) /label=AmpR promoter
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