Basic Vector Information
- Vector Name:
- phis-3GFP3'
- Antibiotic Resistance:
- Ampicillin
- Length:
- 9650 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Bowring FJ, Yeadon PJ, Catcheside DE.
phis-3GFP3' 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 ℃.
phis-3GFP3' vector Sequence
LOCUS 40924_24578 9650 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector phis-3GFP3', complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9650)
AUTHORS Bowring FJ, Yeadon PJ, Catcheside DE.
TITLE Use of fluorescent protein to analyse recombination at three loci in
Neurospora crassa
JOURNAL Fungal Genet. Biol. 49 (8), 619-625 (2012)
PUBMED 22691725
REFERENCE 2 (bases 1 to 9650)
AUTHORS Bowring FJ, Yeadon J, Catcheside DEA.
TITLE Direct Submission
JOURNAL Submitted (11-JUN-2012) School of Biological Sciences, Flinders
University, Sturt Road, Bedford Park, SA 5162, Australia
REFERENCE 3 (bases 1 to 9650)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 9650)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Fungal
Genet. Biol."; date: "2012"; volume: "49"; issue: "8"; pages:
"619-625"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(11-JUN-2012) School of Biological Sciences, Flinders University,
Sturt Road, Bedford Park, SA 5162, Australia"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..9650
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 363..467
/label=AmpR promoter
CDS 468..1325
/label=AmpR
/note="beta-lactamase"
rep_origin 1499..2087
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
promoter 2345..2363
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 2638..2649
/label=Factor Xa site
/note="Factor Xa recognition and cleavage site"
promoter 4008..4026
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
regulatory 4059..4980
/gene="ccg-1"
/label=Neurospora crassa ccg-1 (grg-1) promoter
/note="Neurospora crassa ccg-1 (grg-1) promoter"
/regulatory_class="promoter"
gene 4059..4980
/gene="ccg-1"
/label=ccg-1
primer_bind 4981..4997
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 5002..6904
/function="inactive h1/sgfp fusion as substrate for meiotic
recombination"
/label=3' allele at 6815 renders sgfp non-functional
/note="3' allele at 6815 renders sgfp non-functional"
CDS 6185..6904
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALS*DPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(6925..6941)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
promoter complement(6971..6989)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
misc_feature complement(7532..9633)
/function="targeting to the his-3 locus"
/label=truncated his-3 gene
/note="truncated his-3 gene"
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