Basic Vector Information
pUMRI-13 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 ℃.
pUMRI-13 vector Sequence
LOCUS 40924_45748 5901 bp DNA circular SYN 18-DEC-2018 DEFINITION Cloning vector pUMRI-13, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5901) AUTHORS Xie W, Lv X, Yu H. TITLE pUMRI plasmids for metabolic pathway assembly JOURNAL Unpublished REFERENCE 2 (bases 1 to 5901) AUTHORS Xie W, Lv X, Yu H. TITLE Direct Submission JOURNAL Submitted (21-JUL-2014) Department of Chemical and Biological Engineering, Zhejiang University, Institute of Bioengineering, 34 TianMuShan Road, Hangzhou, Zhejiang 310028, China REFERENCE 3 (bases 1 to 5901) TITLE Direct Submission REFERENCE 4 (bases 1 to 5901) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Unpublished" COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted (21-JUL-2014) Department of Chemical and Biological Engineering, Zhejiang University, Institute of Bioengineering, 34 TianMuShan Road, Hangzhou, Zhejiang 310028, China" COMMENT SGRef: number: 3; type: "Journal Article" COMMENT ##Assembly-Data-START## Assembly Method :: Artificial Sequence v. Artificial Sequence Sequencing Technology :: Artificial Sequence ##Assembly-Data-END## FEATURES Location/Qualifiers source 1..5901 /mol_type="other DNA" /organism="synthetic DNA construct" protein_bind 22..55 /label=loxP /note="Cre-mediated recombination occurs in the 8-bp core sequence (ATGTATGC) (Shaw et al., 2021)." misc_feature 88..1095 /label=GAL1-7 homologous arm /note="GAL1-7 homologous arm" terminator complement(1106..1271) /label=ADH1 terminator /note="transcription terminator for the S. cerevisiae alcohol dehydrogenase 1 (ADH1) gene" CDS complement(1419..1442) /codon_start=1 /label=FLAG /note="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /translation="DYKDDDDK" promoter 1489..2153 /label=GAL1,10 promoter /note="divergent inducible promoter, regulated by Gal4" promoter 2164..2182 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 2200..2229 /codon_start=1 /label=Myc /note="Myc (human c-Myc proto-oncogene) epitope tag" /translation="EQKLISEEDL" terminator 2259..2448 /label=CYC1 terminator /note="transcription terminator for CYC1" promoter complement(2461..2479) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind complement(2502..2535) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." gene 2581..3937 /label=kanMX /note="yeast selectable marker conferring kanamycin resistance (Wach et al., 1994)" promoter 4032..4252 /label=URA3 promoter CDS 4253..5053 /codon_start=1 /label=URA3 /note="orotidine-5'-phosphate decarboxylase, required for uracil biosynthesis" /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN" rep_origin complement(5225..5813) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication"
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