Basic Vector Information
pAW8-yEGFP 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 ℃.
pAW8-yEGFP vector Sequence
LOCUS pAW8-yEGFP. 10655 bp DNA circular SYN 30-NOV-2018
DEFINITION Cloning vector pAW8-yEGFP, complete sequence.
ACCESSION .
VERSION .
KEYWORDS pAW8-yEGFP.
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10655)
AUTHORS Watson AT, Garcia V, Bone N, Carr AM, Armstrong J.
TITLE Gene tagging and gene replacement using recombinase-mediated
cassette exchange in Schizosaccharomyces pombe
JOURNAL Gene 407 (1-2), 63-74 (2008)
PUBMED 18054176
REFERENCE 2 (bases 1 to 10655)
AUTHORS Watson AT.
TITLE Direct Submission
JOURNAL Submitted (13-AUG-2007) Genome Damage and Stability Centre,
University of Sussex, Falmer, Brighton, East Sussex BN1 9RQ, UK
REFERENCE 3 (bases 1 to 10655)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 10655)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Gene 407
(1-2), 63-74 (2008)"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(13-AUG-2007) Genome Damage and Stability Centre, University of
Sussex, Falmer, Brighton, East Sussex BN1 9RQ, UK"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..10655
/mol_type="other DNA"
/organism="synthetic DNA construct"
rep_origin 575..1356
/label=ars1
/note="Schizosaccharomyces pombe autonomously replicating
sequence ars1"
promoter 1716..1820
/label=AmpR promoter
CDS 1821..2678
/label=AmpR
/note="beta-lactamase"
rep_origin 2852..3440
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 3728..3749
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 3764..3794
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3802..3818
/label=lac repressor encoded by lacI binding site
/bound_moiety="lac repressor encoded by lacI"
/note="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 3826..3842
/label=M13 rev
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
gene complement(3976..5332)
/label=kanMX
/note="yeast selectable marker conferring kanamycin
resistance (Wach et al., 1994)"
terminator complement(5407..5594)
/label=ADH1 terminator
/note="transcription terminator for the S. cerevisiae
alcohol dehydrogenase 1 (ADH1) gene"
CDS complement(5618..6331)
/label=GFP
/note="green fluorescent protein"
protein_bind 6340..6373
/label=loxP
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
primer_bind complement(6389..6405)
/label=M13 fwd
/note="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
regulatory 6612..7168
/label=nmt1
/note="nmt1"
/regulatory_class="terminator"
CDS complement(7177..8166)
/label=Cre
/note="site-specific recombinase"
intron complement(8167..8215)
/gene="cre"
/note="Schizosaccharomyces pombe rad50 intron 1"
regulatory complement(8255..8865)
/label=nmt41
/note="nmt41"
/regulatory_class="promoter"
promoter 8884..9303
/label=LEU2 promoter
CDS 9304..10395
/label=LEU2
/note="3-isopropylmalate dehydrogenase, required for
leucine biosynthesis"
regulatory 10399..10655
/gene="LEU2"
/regulatory_class="terminator"
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