pAW8-yEGFP vector (V010153)

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"