LOCUS       Exported                5785 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  PCR template vector pUG30, complete sequence.
ACCESSION   AF298781
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5785)
  AUTHORS   Gueldener U, Hegemann JH.
  TITLE     A second generation of GFP-vectors for subcelluar localization 
            studies in budding yeast
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5785)
  AUTHORS   Gueldener U, Hegemann JH.
  TITLE     Direct Submission
  JOURNAL   Submitted (23-AUG-2000) Institut fuer Mikrobiologie, 
            Heinrich-Heine-Universitaet, Universtitaetsstr. 1, Duesseldorf 
            40225, Germany
REFERENCE   3  (bases 1 to 5785)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5785
                     /organism="PCR template vector pUG30"
                     /mol_type="other DNA"
                     /label=derived from Saccharomyces cerevisiae
                     /note="derived from Saccharomyces cerevisiae"
                     /db_xref="taxon:142961"
     promoter        103..133
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    141..157
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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     165..181
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        202..220
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     regulatory      complement(234..626)
                     /regulatory_class="promoter"
                     /note="from MET25"
     promoter        271..620
                     /gene="S. cerevisiae MET17"
                     /gene_synonym="MET15; MET25"
                     /label=MET17 promoter
                     /note="expression is repressed by methionine"
     primer_bind     621..637
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             676..1392
                     /codon_start=1
                     /gene="EGFP3"
                     /product="enhanced green fluorescent protein 3"
                     /label=EGFP3
                     /protein_id="AAG34511.1"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     gene            676..1392
                     /gene="EGFP3"
                     /label=EGFP3
     regulatory      complement(1410..1668)
                     /regulatory_class="terminator"
                     /note="from CYC1"
     terminator      1411..1658
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     promoter        complement(1677..1695)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1705..1721)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1829..1862
                     /label=loxP site
                     /note="loxP site"
     protein_bind    complement(1829..1862)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     regulatory      1863..2260
                     /regulatory_class="promoter"
                     /note="from TEF"
     gene            1917..3273
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        1917..2260
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             2261..3070
                     /codon_start=1
                     /gene="kanMX"
                     /product="geneticin resistance protein"
                     /label=kanMX
                     /protein_id="AAG34512.1"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     gene            2261..3070
                     /gene="kanMX"
                     /label=kanMX
     regulatory      3071..3335
                     /regulatory_class="terminator"
                     /note="from TEF"
     terminator      3076..3273
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     misc_feature    3336..3369
                     /label=loxP site
                     /note="loxP site"
     protein_bind    complement(3336..3369)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     promoter        complement(3423..3441)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      complement(3699..4287)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4458..5318)
                     /codon_start=1
                     /gene="AmpR"
                     /product="ampicillin resistance protein"
                     /label=AmpR
                     /protein_id="AAG34513.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(4458..5318)
                     /gene="AmpR"
                     /label=AmpR
     promoter        complement(5319..5423)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5769..2
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"