LOCUS       Exported                4881 bp ds-DNA    circular SYN 29-11-2013
DEFINITION  Plasmid with a TRP1 marker for swapping in the GAL1 promoter and 
            adding a GFP tag.
ACCESSION   .
VERSION     .
KEYWORDS    pFA6a-TRP1-PGAL1-GFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4881)
  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 4881)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4881
                     /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"
     promoter        1542..1689
                     /gene="S. cerevisiae TRP1"
                     /note="TRP1 promoter"
     CDS             1690..2364
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /note="TRP1"
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     promoter        complement(2519..2537)
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      complement(2795..3383)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number colE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3554..4414)
                     /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(4415..4519)
                     /gene="bla"
                     /note="AmpR promoter"
     promoter        4865..2
                     /note="SP6 promoter"
                     /note="promoter for bacteriophage SP6 RNA polymerase"