Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V002334 | pUG34 | In stock, instant shipping |
Buy one, get one free! |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
- Vector Name:
- pUG34
- Antibiotic Resistance:
- Ampicillin
- Length:
- 6301 bp
- Type:
- N-terminal GFP fusion vector
- Replication origin:
- ori
- Source/Author:
- Gueldener U, Hegemann JH.
- Promoter:
- MET17
- Growth Strain(s):
- DH10B
- Growth Temperature:
- 37℃
pUG34 vector Map
Plasmid 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 ℃.
6. The concentration of plasmid re-measurement sometimes differs from the nominal value, which may be due to the position of the lyophilized plasmid in the tube, the efficiency of the re-dissolution, the measurement bias, and adsorption on the wall of the tube, therefore, it is recommended to transform and extract the plasmid before using it
General Plasmid Transform Protocol
1. Take one 100μl of the competent cells and thaw it on ice for 10min, add 2μl of plasmid, then ice bath for 30min, then heat-shock it at 42℃ for 60s, do not stir, and then ice bath for 2min.
2. Add 900μl of LB liquid medium without antibiotics, and incubate at 37℃ for 45min (30℃ for 1-1.5 hours) with 180rpm shaking.
3. Centrifuge at 6000rpm for 5min, leave only 100μl of supernatant to resuspend the bacterial precipitate and spread it onto the target plasmid-resistant LB plate.
4. Invert the plate and incubate at 37℃ for 14h, or at 30℃ for 20h.
5. Pick a single colony into LB liquid medium, add the corresponding antibiotics, incubate at 220rpm for 14h, and extract the plasmid according to the experimental needs and the instructions of the plasmid extraction kit.
pUG34 vector Sequence
LOCUS Exported 6301 bp DNA circular SYN 23-OCT-2024
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 6301)
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JOURNAL Unpublished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 6301)
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 6301)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 6301)
TITLE Direct Submission
REFERENCE 5 (bases 1 to 6301)
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"
COMMENT SGRef: number: 4; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..6301
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 317..504
/label=HIS3 promoter
CDS 505..1164
/codon_start=1
/label=HIS3
/note="imidazoleglycerol-phosphate dehydratase, required
for histidine biosynthesis"
/translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
KEALGAVRGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHF
LESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
rep_origin complement(1427..1882)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind 2027..2043
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 2053..2071
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
terminator complement(2090..2337)
/label=CYC1 terminator
/note="transcription terminator for CYC1"
primer_bind 2340..2356
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2390..2406)
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
CDS complement(2407..3120)
/codon_start=1
/label=yeGFP
/note="yeast-enhanced green fluorescent protein"
/translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
FVTAAGITHGMDELYK"
promoter complement(3127..3476)
/label=MET17 promoter
/note="expression is repressed by methionine"
promoter complement(3527..3545)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(3566..3582)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(3590..3606)
/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(3614..3644)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(3659..3680)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(3968..4556)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4730..5587)
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(5588..5692)
/label=AmpR promoter
misc_feature 5729..6232
/label=CEN/ARS
/note="S. cerevisiae CEN6 centromere fused to an
autonomously replicating sequence"