Basic Vector Information
P416 GPD 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 ℃.
P416 GPD vector Sequence
LOCUS 40924_2792 5778 bp DNA circular SYN 13-JAN-2022 DEFINITION synthetic circular DNA. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5778) TITLE Direct Submission REFERENCE 2 (bases 1 to 5778) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article" FEATURES Location/Qualifiers source 1..5778 /mol_type="other DNA" /organism="synthetic DNA construct" misc_feature complement(70..573) /label=CEN/ARS /note="S. cerevisiae CEN6 centromere fused to an autonomously replicating sequence" promoter 610..714 /label=AmpR promoter CDS 715..1572 /codon_start=1 /label=AmpR /note="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" rep_origin 1746..2334 /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" protein_bind 2622..2643 /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." promoter 2658..2688 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2696..2712 /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)." primer_bind 2720..2736 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" promoter 2757..2775 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" promoter 2800..3443 /label=GAP promoter /note="promoter for glyceraldehyde-3-phosphate dehydrogenase; also known as the TDH3 promoter" primer_bind 3449..3465 /label=SK primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3499..3515) /label=KS primer /note="common sequencing primer, one of multiple similar variants" terminator 3518..3765 /label=CYC1 terminator /note="transcription terminator for CYC1" promoter complement(3784..3802) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(3812..3828) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" rep_origin 3973..4428 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" CDS complement(4562..5362) /codon_start=1 /label=URA3 /note="orotidine-5'-phosphate decarboxylase, required for uracil biosynthesis" /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN" promoter complement(5363..5583) /label=URA3 promoter
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