LOCUS       Exported                8197 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pTL85, complete sequence.
ACCESSION   FJ834450
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8197)
  AUTHORS   Baruffini E, Serafini F, Lodi T.
  TITLE     Construction and characterization of centromeric, episomal and 
            GFP-containing vectors for Saccharomyces cerevisiae prototrophic 
            strains
  JOURNAL   J. Biotechnol. 143 (4), 247-254 (2009)
  PUBMED    19683551
REFERENCE   2  (bases 1 to 8197)
  AUTHORS   Baruffini E, Serafini F, Lodi T.
  TITLE     Direct Submission
  JOURNAL   Submitted (18-MAR-2009) Department of Genetics, Biology of 
            Microorganisms, Anthropology, Evolution, University of Parma, Viale 
            Usberti 11/A, Parma 43100, Italy
REFERENCE   3  (bases 1 to 8197)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8197
                     /organism="Expression vector pTL85"
                     /lab_host="Saccharomyces cerevisiae"
                     /mol_type="other DNA"
                     /db_xref="taxon:670505"
     protein_bind    106..127
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        142..172
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    180..196
                     /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     204..220
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      249..496
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     CDS             complement(506..1222)
                     /codon_start=1
                     /gene="yEGFP"
                     /product="green fluorescent protein"
                     /label=yEGFP
                     /note="optimized for yeast codon usage"
                     /protein_id="ACV50017.1"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     gene            complement(506..1222)
                     /gene="yEGFP"
                     /label=yEGFP
     regulatory      complement(1223..1673)
                     /regulatory_class="promoter"
                     /note="MET17 promoter"
     primer_bind     1225..1241
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1231..1287
                     /label=multiple cloning site for GFP fusion
                     /note="multiple cloning site for GFP fusion"
     primer_bind     complement(1275..1291)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(1292..1641)
                     /gene="S. cerevisiae MET17"
                     /gene_synonym="MET15; MET25"
                     /label=MET17 promoter
                     /note="expression is repressed by methionine"
     primer_bind     complement(1686..1702)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      2139..3485
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     rep_origin      2140..3305
                     /label=2 micron origin
                     /note="2 micron origin"
     regulatory      3592..3982
                     /regulatory_class="promoter"
                     /note="TEF-promoter"
     gene            3639..4995
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        3639..3982
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             3983..4792
                     /codon_start=1
                     /gene="kanR"
                     /product="KanR"
                     /label=kanR
                     /note="aminoglycoside 3-phosphotransferase; APH"
                     /protein_id="ACV50018.1"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     gene            3983..4792
                     /gene="kanR"
                     /label=kanR
     regulatory      4790..5024
                     /regulatory_class="terminator"
                     /note="TEF-terminator"
     terminator      4798..4995
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     promoter        6291..6395
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6396..7256
                     /codon_start=1
                     /gene="ampR"
                     /product="AmpR"
                     /label=ampR
                     /note="beta-lactamase"
                     /protein_id="ACV50019.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            6396..7256
                     /gene="ampR"
                     /label=ampR
     rep_origin      7427..8015
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"