phis-5GFP vector (V005555)

Basic Vector Information

      • Vector Name:
      • phis-5GFP
      • Antibiotic Resistance:
      • Ampicillin
      • Length:
      • 9649 bp
      • Type:
      • Cloning vector
      • Source/Author:
      • Bowring FJ, Yeadon PJ, Catcheside DE.

phis-5GFP vector Vector Map

phis-5GFP9649 bp4008001200160020002400280032003600400044004800520056006000640068007200760080008400880092009600CAP binding sitelac promoterlac operatorM13 revSP6 promotertruncated his-5 geneNeurospora crassa ccg-1 (grg-1) promoterSK primerEGFP3' flank ofNeurospora crassa his-5 geneT7 promoterM13 fwdf1 oriNeoR/KanRAmpRori

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

Copy Sequence

Download GeneBank File(.gb)

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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