LOCUS       Exported                7933 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Integrating expression vector pARC25B, complete sequence.
ACCESSION   AF359244
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7933)
  AUTHORS   Tugendreich S, Perkins E, Couto J, Barthmaier P, Sun D, Tang S, 
            Tulac S, Nguyen A, Yeh E, Mays A, Wallace E, Lila T, Shivak D, 
            Prichard M, Andrejka L, Kim R, Melese T.
  TITLE     A streamlined process to phenotypically profile heterologous cDNAs 
            in parallel using yeast cell-based assays
  JOURNAL   Genome Res. 11 (11), 1899-1912 (2001)
  PUBMED    11691855
REFERENCE   2  (bases 1 to 7933)
  AUTHORS   Tugendreich SM, Perkins E, Couto J, Barthmaier P, Sun D, Tang S, 
            Tulac S, Nguyen A, Yeh E, Mays A, Lila T, Shivak D, Prichard M, 
            Andrejka L, Kim R, Melese T.
  TITLE     Direct Submission
  JOURNAL   Submitted (09-MAR-2001) Surrogate Genetics, Iconix Pharmaceuticals, 
            Inc., 850 Maude Ave, Mountain View, CA 94043, USA
REFERENCE   3  (bases 1 to 7933)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7933
                     /organism="Integrating expression vector pARC25B"
                     /lab_host="Saccharomyces cerevisiae"
                     /mol_type="other DNA"
                     /label=episomal
                     /note="episomal"
                     /db_xref="taxon:156893"
     CDS             complement(663..1769)
                     /codon_start=1
                     /product="LEU2 marker"
                     /label=LEU2 marker
                     /protein_id="AAK31262.1"
                     /translation="MSAPMSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDF
                     ENHLIGGAAIDATGVPLPDEALEASKKVDAVLLGAVAGPKWGTGSVRPEQGLLKIRKEL
                     QLYANLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDS
                     EQYTVPEVQRITRMAAFMALQHEPPLPIWSLDKANLLASSRLWRKTVEETIKNEFPTLK
                     VQHQLIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDK
                     NTAFGLYEPCHGSAPDLPKNKVDPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAG
                     IRTGDLGGSNSTTEVGDAVAEEVKKILA"
     CDS             complement(663..1757)
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKVDAVLLGAVAGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANLLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVDPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     promoter        complement(1770..2174)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     rep_origin      complement(2474..2929)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     3074..3090
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3100..3118
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(3442..3883)
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    3766..3883
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     promoter        complement(4206..4224)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4245..4261)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4269..4285
                     /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(4293..4323)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     rep_origin      complement(4647..5235)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5406..6266)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6267..6371)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6460..7802
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(7423..7470)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."