Basic Vector Information
- Vector Name:
- pUG34
- Antibiotic Resistance:
- Ampicillin
- Length:
- 6297 bp
- Type:
- N-terminal GFP fusion vector
- Replication origin:
- ori
- Source/Author:
- Gueldener U, Hegemann JH.
- Promoter:
- MET17
pUG34 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 ℃.
pUG34 vector Sequence
LOCUS 40924_45488 6297 bp DNA circular SYN 18-DEC-2018
DEFINITION N-terminal GFP fusion vector pUG34, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6297)
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JOURNAL Unpublished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 6297)
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 6297)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 6297)
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"Unpublished"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..6297
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 317..503
/label=HIS3 promoter
CDS 504..1160
/codon_start=1
/label=HIS3
/note="imidazoleglycerol-phosphate dehydratase, required
for histidine biosynthesis"
/translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
KEALLARGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHFL
ESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
rep_origin complement(1423..1878)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind 2023..2039
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 2049..2067
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
terminator complement(2086..2333)
/label=CYC1 terminator
/note="transcription terminator for CYC1"
primer_bind 2336..2352
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2386..2402)
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
CDS complement(2403..3116)
/codon_start=1
/label=yeGFP
/note="yeast-enhanced green fluorescent protein"
/translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
FVTAAGITHGMDELYK"
promoter complement(3123..3472)
/label=MET17 promoter
/note="expression is repressed by methionine"
promoter complement(3523..3541)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(3562..3578)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(3586..3602)
/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(3610..3640)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(3655..3676)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(3964..4552)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4726..5583)
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(5584..5688)
/label=AmpR promoter
misc_feature 5725..6228
/label=CEN/ARS
/note="S. cerevisiae CEN6 centromere fused to an
autonomously replicating sequence"
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