LOCUS       Exported                5404 bp ds-DNA    circular SYN 24-12-2013
DEFINITION  Plasmid with a kanMX marker for swapping in the GAL1 promoter and 
            adding a GFP tag.
ACCESSION   .
VERSION     .
KEYWORDS    pFA6a-kanMX6-PGAL1-GFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5404)
  AUTHORS   Longtine MS, McKenzie A, Demarini DJ, Shah NG, Wach A, Brachat A, 
            Philippsen P, Pringle JR.
  TITLE     Additional modules for versatile and economical PCR-based gene 
            deletion and modification in Saccharomyces cerevisiae.
  JOURNAL   Yeast 1998;14:953-61.
  PUBMED    9717241
REFERENCE   2  (bases 1 to 5404)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5404
                     /organism="synthetic DNA construct"
                     /lab_host="Saccharomyces cerevisiae"
                     /mol_type="other DNA"
     terminator      50..237
                     /note="ADH1 terminator"
     CDS             complement(258..970)
                     /codon_start=3
                     /gene="gfp"
                     /product="green fluorescent protein, S65T variant"
                     /note="GFP(S65T)"
                     /db_xref="GI:1019894"
                     /protein_id="AAA79306.1"
                     /translation="KGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFI
                     CTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGNY
                     KTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNF
                     KIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFV
                     TAAGITHGMDELYK"
     CDS             complement(984..986)
                     /codon_start=1
                     /note="start codon"
                     /translation="M"
     promoter        complement(1006..1447)
                     /gene="S. cerevisiae GAL1"
                     /note="GAL1 promoter"
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    1330..1447
                     /bound_moiety="Gal4"
                     /note="UAS"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     gene            1581..2937
                     /note="kanMX"
                     /note="yeast selectable marker conferring kanamycin 
                     resistance"
     promoter        1581..1924
                     /note="TEF promoter"
                     /note="Ashbya gossypii TEF promoter"
     CDS             1925..2734
                     /codon_start=1
                     /gene="aph(3)-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /note="KanR"
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     terminator      2740..2937
                     /note="TEF terminator"
                     /note="Ashbya gossypii TEF terminator"
     promoter        complement(3042..3060)
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      complement(3318..3906)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number colE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4077..4937)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4938..5042)
                     /gene="bla"
                     /note="AmpR promoter"
     promoter        5388..2
                     /note="SP6 promoter"
                     /note="promoter for bacteriophage SP6 RNA polymerase"