Basic Vector Information
phis-5GFP 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-5GFP vector Sequence
LOCUS 40924_24583 9649 bp DNA circular SYN 18-DEC-2018 DEFINITION Cloning vector phis-5GFP, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9649) 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 9649) 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 9649) TITLE Direct Submission REFERENCE 4 (bases 1 to 9649) 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..9649 /mol_type="other DNA" /organism="synthetic DNA construct" protein_bind 107..128 /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." promoter 143..173 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 181..197 /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 205..221 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" promoter 239..257 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" misc_feature 336..1578 /function="targeting to the his-5 locus" /label=truncated his-5 gene /note="truncated his-5 gene" regulatory 1640..2561 /gene="ccg-1" /label=Neurospora crassa ccg-1 (grg-1) promoter /note="Neurospora crassa ccg-1 (grg-1) promoter" /regulatory_class="promoter" gene 1640..2561 /gene="ccg-1" /label=ccg-1 primer_bind 2562..2578 /label=SK primer /note="common sequencing primer, one of multiple similar variants" CDS 3766..4482 /label=EGFP /note="enhanced GFP" misc_feature 4494..6013 /function="targeting to the his-5 locus" /label=3' flank ofNeurospora crassa his-5 gene /note="3' flank ofNeurospora crassa his-5 gene" promoter complement(6083..6101) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(6108..6124) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" rep_origin 6265..6693 /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" CDS 7037..7828 /label=NeoR/KanR /note="aminoglycoside phosphotransferase" CDS 7849..8706 /label=AmpR /note="beta-lactamase" rep_origin 8880..9468 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication"
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