Basic Vector Information
pUG35 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 ℃.
pUG35 vector Sequence
LOCUS 40924_45493 6231 bp DNA circular SYN 18-DEC-2018 DEFINITION C-terminal GFP fusion vector pUG35, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6231) AUTHORS Gueldener U, Hegemann JH. TITLE A second generation of GFP-vectors for subcelluar localization studies in budding yeast JOURNAL Unpublished REFERENCE 2 (bases 1 to 6231) AUTHORS Gueldener U, Hegemann JH. TITLE Direct Submission JOURNAL Submitted (23-AUG-2000) Institut fuer Mikrobiologie, Heinrich-Heine-Universitaet, Universtitaetsstr. 1, Duesseldorf 40225, Germany REFERENCE 3 (bases 1 to 6231) TITLE Direct Submission REFERENCE 4 (bases 1 to 6231) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Unpublished" COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted (23-AUG-2000) Institut fuer Mikrobiologie, Heinrich-Heine-Universitaet, Universtitaetsstr. 1, Duesseldorf 40225, Germany" COMMENT SGRef: number: 3; type: "Journal Article" FEATURES Location/Qualifiers source 1..6231 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 196..416 /label=URA3 promoter CDS 417..1217 /codon_start=1 /label=URA3 /note="orotidine-5'-phosphate decarboxylase, required for uracil biosynthesis" /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN" rep_origin complement(1351..1806) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind 1951..1967 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" promoter 1977..1995 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" terminator complement(2014..2261) /label=CYC1 terminator /note="transcription terminator for CYC1" CDS complement(2274..2987) /codon_start=1 /label=yeGFP /note="yeast-enhanced green fluorescent protein" /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE FVTAAGITHGMDELYK" primer_bind 2990..3006 /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3040..3056) /label=SK primer /note="common sequencing primer, one of multiple similar variants" promoter complement(3057..3406) /label=MET17 promoter /note="expression is repressed by methionine" promoter complement(3457..3475) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(3496..3512) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind complement(3520..3536) /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(3544..3574) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind complement(3589..3610) /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(3898..4486) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(4660..5517) /codon_start=1 /label=AmpR /note="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(5518..5622) /label=AmpR promoter misc_feature 5659..6162 /label=CEN/ARS /note="S. cerevisiae CEN6 centromere fused to an autonomously replicating sequence"
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