LOCUS       Exported                6934 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector p413-TR-SSRE-GFP, complete sequence.
ACCESSION   DQ232596
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6934)
  AUTHORS   Chen M-T., Weiss R.
  TITLE     Artificial cell-cell communication in yeast Saccharomyces cerevisiae
            using signaling elements from Arabidopsis thaliana
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6934)
  AUTHORS   Chen M-T., Weiss R.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-OCT-2005) Electrical Engineering, Princeton 
            University, Engineering Quadrangle, Olden Street, Princeton, NJ 
            08544, USA
REFERENCE   3  (bases 1 to 6934)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6934
                     /organism="Expression vector p413-TR-SSRE-GFP"
                     /mol_type="other DNA"
                     /db_xref="taxon:356369"
     promoter        317..503
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             504..1163
                     /codon_start=1
                     /gene="HIS3"
                     /product="HIS3"
                     /label=HIS3
                     /note="imidazoleglycerol-phosphate dehydratase"
                     /protein_id="ABB83380.1"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALLARGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHFL
                     ESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     gene            504..1163
                     /gene="HIS3"
                     /label=HIS3
     rep_origin      complement(1423..1878)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     2023..2039
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2049..2067
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      complement(2409..2637)
                     /regulatory_class="terminator"
                     /note="ADH1 terminator"
     terminator      2438..2625
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     CDS             complement(2646..3362)
                     /codon_start=1
                     /product="yEGFP3"
                     /label=yEGFP3
                     /note="codon-optimized GFP containing S65G and S72A 
                     mutations; similar to GenBank Accession Number U73901"
                     /protein_id="ABB83382.1"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     regulatory      complement(3382..4114)
                     /regulatory_class="promoter"
                     /note="TR-SSRE promoter"
     primer_bind     complement(4124..4140)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(4160..4178)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4199..4215)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4223..4239
                     /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)."
     promoter        complement(4247..4277)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     rep_origin      complement(4601..5189)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5360..6220)
                     /codon_start=1
                     /gene="ampR"
                     /product="AmpR"
                     /label=ampR
                     /note="beta-lactamase; ampR ampicillin resistance"
                     /protein_id="ABB83381.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(5360..6220)
                     /gene="ampR"
                     /label=ampR
     promoter        complement(6221..6325)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    6362..6865
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"