Basic Vector Information
pUDE152 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 ℃.
pUDE152 vector Sequence
LOCUS 40924_45433 8107 bp DNA circular SYN 18-DEC-2018 DEFINITION Expression vector pUDE152, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8107) AUTHORS Kozak BU, van Rossum HM, Benjamin KR, Wu L, Daran JM, Pronk JT, van Maris AJ. TITLE Replacement of the Saccharomyces cerevisiae acetyl-CoA synthetases by alternative pathways for cytosolic acetyl-CoA synthesis JOURNAL Metab. Eng. (2013) In press PUBMED 24269999 REFERENCE 2 (bases 1 to 8107) AUTHORS Kozak BU, van Rossum HM, Benjamin KR, Wu L, Daran J-M.G., Pronk JT, van Maris AJA. TITLE Direct Submission JOURNAL Submitted (31-MAY-2013) Biotechnology, Delft University of Technology, Julianalaan 67, Delft 2628BC, The Netherlands REFERENCE 3 (bases 1 to 8107) TITLE Direct Submission REFERENCE 4 (bases 1 to 8107) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Metab. Eng. (2013) In press" COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted (31-MAY-2013) Biotechnology, Delft University of Technology, Julianalaan 67, Delft 2628BC, The Netherlands" COMMENT SGRef: number: 3; type: "Journal Article" FEATURES Location/Qualifiers source 1..8107 /mol_type="other DNA" /organism="synthetic DNA construct" rep_origin complement(183..771) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(945..1802) /codon_start=1 /label=AmpR /note="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(1803..1907) /label=AmpR promoter rep_origin complement(1934..3276) /direction=LEFT /label=2u ori /note="yeast 2u plasmid origin of replication" promoter 3540..3760 /label=URA3 promoter CDS 3761..4561 /codon_start=1 /label=URA3 /note="orotidine-5'-phosphate decarboxylase, required for uracil biosynthesis" /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN" rep_origin complement(4695..5150) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind 5295..5311 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" promoter 5321..5339 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" terminator complement(5358..5605) /label=CYC1 terminator /note="transcription terminator for CYC1" primer_bind 5608..5624 /label=KS primer /note="common sequencing primer, one of multiple similar variants" protein_bind 5657..5681 /label=attB2 /note="recombination site for the Gateway(R) BP reaction" CDS complement(5697..7106) /codon_start=1 /gene="lin1129" /product="lin1129" /label=lin1129 /protein_id="AHB33301.1" /translation="MESLELEQLVKKVLLEKLAEQKEVPTKTTTQGAKSGVFDTVDEAV QAAVIAQNCYKEKSLEERRNVVKAIREALYPEIETIATRAVAETGMGNVTDKILKNTLA IEKTPGVEDLYTEVATGDNGMTLYELSPYGVIGAVAPSTNPTETLICNSIGMLAAGNAV FYSPHPGAKNISLWLIEKLNTIVRDSCGIDNLIVTVAKPSIQAAQEMMNHPKVPLLVIT GGPGVVLQAMQSGKKVIGAGAGNPPSIVDETANIEKAAADIVDGASFDHNILCIAEKSV VAVDSIADFLLFQMEKNGALHVTNPSDIQKLEKVAVTDKGVTNKKLVGKSATEILKEAG IACDFTPRLIIVETEKSHPFATVELLMPIVPVVRVPDFDEALEVAIELEQGLHHTATMH SQNISRLNKAARDMQTSIFVKNGPSFAGLGFRGEGSTTFTIATPTGEGTTTARHFARRR RCVLTDGFSIR" gene complement(5697..7106) /gene="lin1129" /label=lin1129 protein_bind complement(7122..7146) /label=attB1 /note="recombination site for the Gateway(R) BP reaction" primer_bind complement(7154..7170) /label=SK primer /note="common sequencing primer, one of multiple similar variants" promoter complement(7176..7820) /label=GAP promoter /note="promoter for glyceraldehyde-3-phosphate dehydrogenase; also known as the TDH3 promoter" promoter complement(7849..7867) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(7888..7904) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind complement(7912..7928) /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(7936..7966) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind complement(7981..8002) /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP."
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