Basic Vector Information
pUMRI-11 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-11 vector Sequence
LOCUS 40924_45738 5859 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pUMRI-11, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5859)
AUTHORS Xie W, Lv X, Yu H.
TITLE pUMRI plasmids for metabolic pathway assembly
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 5859)
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 5859)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 5859)
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"Unpublished"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..5859
/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 113..1029
/label=DPP1 homologous arm
/note="DPP1 homologous arm"
terminator complement(1064..1229)
/label=ADH1 terminator
/note="transcription terminator for the S. cerevisiae
alcohol dehydrogenase 1 (ADH1) gene"
CDS complement(1377..1400)
/codon_start=1
/label=FLAG
/note="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/translation="DYKDDDDK"
promoter 1447..2111
/label=GAL1,10 promoter
/note="divergent inducible promoter, regulated by Gal4"
promoter 2122..2140
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 2158..2187
/codon_start=1
/label=Myc
/note="Myc (human c-Myc proto-oncogene) epitope tag"
/translation="EQKLISEEDL"
terminator 2217..2406
/label=CYC1 terminator
/note="transcription terminator for CYC1"
promoter complement(2419..2437)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
misc_feature complement(2460..2493)
/label=Loxp site
/note="Loxp site"
protein_bind complement(2460..2493)
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
gene 2539..3895
/label=kanMX
/note="yeast selectable marker conferring kanamycin
resistance (Wach et al., 1994)"
promoter 3990..4210
/label=URA3 promoter
CDS 4211..5011
/codon_start=1
/label=URA3
/note="orotidine-5'-phosphate decarboxylase, required for
uracil biosynthesis"
/translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
rep_origin complement(5183..5771)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
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