Basic Vector Information
pUG30 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 ℃.
pUG30 vector Sequence
LOCUS 40924_45483 5785 bp DNA circular SYN 18-DEC-2018 DEFINITION PCR template vector pUG30, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5785) 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 TITLE A second generation of GFP-vectors for subcelluar localization studies in budding yeast JOURNAL Unpublished REFERENCE 2 (bases 1 to 5785) AUTHORS Gueldener U, Hegemann JH.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 5785) TITLE Direct Submission REFERENCE 4 (bases 1 to 5785) 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 TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Unpublished" COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted (23-AUG-2000) Institut fuer Mikrobiologie, Heinrich-Heine-Universitaet,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..5785 /mol_type="other DNA" /organism="synthetic DNA construct" protein_bind 67..88 /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." promoter 103..133 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 141..157 /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 165..181 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" promoter 202..220 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" promoter 271..620 /label=MET17 promoter /note="expression is repressed by methionine" primer_bind 621..637 /label=SK primer /note="common sequencing primer, one of multiple similar variants" CDS 676..1389 /codon_start=1 /label=yeGFP /note="yeast-enhanced green fluorescent protein" /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE FVTAAGITHGMDELYK" terminator 1411..1658 /label=CYC1 terminator /note="transcription terminator for CYC1" promoter complement(1677..1695) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(1705..1721) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" protein_bind 1829..1862 /label=loxP /note="Cre-mediated recombination occurs in the 8-bp core sequence (ATGTATGC) (Shaw et al., 2021)." gene 1917..3273 /label=kanMX /note="yeast selectable marker conferring kanamycin resistance (Wach et al., 1994)" misc_feature 3336..3369 /label=loxP site /note="loxP site" protein_bind complement(3336..3369) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." promoter complement(3423..3441) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" rep_origin complement(3699..4287) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(4461..5318) /codon_start=1 /label=AmpR /note="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(5319..5423) /label=AmpR promoter promoter 5769..5785 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase"
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