LOCUS       Exported                5074 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pFA-LIFEACT-GFP-URA3, complete sequence.
ACCESSION   JN051499
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5074)
  AUTHORS   Epp E, Nazarova E, Douglas LM, Konopka JB, Vogel J, Whiteway M.
  TITLE     Actin dynamics and endocytosis in the absence of Arp2/3 in Candida 
            albicans
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5074)
  AUTHORS   Epp E, Nazarova E, Douglas LM, Konopka JB, Vogel J, Whiteway M.
  TITLE     Direct Submission
  JOURNAL   Submitted (24-MAY-2011) Biology, McGill University, 1205 Docteur 
            Penfield, Montreal, Quebec H3A 1B1, Canada
REFERENCE   3  (bases 1 to 5074)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5074
                     /organism="Cloning vector pFA-LIFEACT-GFP-URA3"
                     /mol_type="other DNA"
                     /label=derived from Candida albicans
                     /note="derived from Candida albicans"
                     /db_xref="taxon:1043207"
     gene            51..939
                     /gene="LIFEACT-GFP"
                     /label=LIFEACT-GFP
                     /note="LIFEACT coupled to green fluorescent protein for C. 
                     albicans; integration at the actin locus by homologous 
                     recombination through upstream and downstream"
     CDS             151..201
                     /codon_start=1
                     /product="peptide that binds filamentous actin (Riedl et 
                     al., 2008)"
                     /label=Lifeact
                     /note="first 17 amino acids of budding yeast Abp140"
                     /translation="MGVADLIKKFESISKEE"
     CDS             226..939
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTLCYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKAN
                     FKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     regulatory      940..1195
                     /regulatory_class="terminator"
                     /note="Saccharomyces cerevisiae URA3 terminator"
     CDS             1598..2410
                     /codon_start=1
                     /gene="URA3"
                     /product="Ura3"
                     /label=URA3
                     /protein_id="AEI30141.1"
                     /translation="MTVNTKTYSERAETHASPVAQRLFRLMELKKTNLCASIDVDTTKE
                     FLELIDKLGPYVCLIKTHIDIINDFSYESTIEPLLELSRKHQFMIFEDRKFADIGNTVK
                     KQYIGGVYKISSWADITNAHGVTGNGVVEGLKQGAKETTTNQEPRGLLMLAELSSVGSL
                     AYGEYSQKTVEIAKSDKEFVIGFIAQRDMGGQEEGFDWLIMTPGVGLDDKGDGLGQQYR
                     TVDEVVSTGTDIIIVGRGLFGKGRDPDIEGKRYRNAGWNAYLKKTGQL"
     gene            1598..2410
                     /gene="URA3"
                     /label=URA3
                     /note="selectable marker URA3 from C. albicans"
     promoter        complement(2712..2730)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      complement(2988..3576)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3747..4607)
                     /codon_start=1
                     /gene="bla"
                     /product="beta lactamase"
                     /label=bla
                     /protein_id="AEI30142.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(3747..4607)
                     /gene="bla"
                     /label=bla
     promoter        complement(4608..4712)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5058..2
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"