LOCUS       40924_4724        7933 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Integrating expression vector pARC25B, complete sequence.
ACCESSION   .
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)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7933)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Genome 
            Res."; date: "2001"; volume: "11"; issue: "11"; pages: "1899-1912"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (09-MAR-2001) Surrogate Genetics, Iconix Pharmaceuticals, Inc., 850 
            Maude Ave, Mountain View, CA 94043, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7933
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(666..1757)
                     /codon_start=1
                     /label=LEU2
                     /note="3-isopropylmalate dehydrogenase, required for
                     leucine biosynthesis"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKVDAVLLGAVAGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANLLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVDPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     promoter        complement(1770..2174)
                     /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(3441..4087)
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
     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    complement(4269..4285)
                     /label=lac operator
                     /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"
     protein_bind    complement(4338..4359)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4647..5235)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5409..6266)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6267..6371)
                     /label=AmpR promoter
     rep_origin      complement(6460..7802)
                     /direction=LEFT
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatcga ctacgtcgta aggccgtttc tgacagagta aaattcttga gggaactttc
      241 accattatgg gaaatgcttc aagaaggtat tgacttaaac tccatcaaat ggtcaggtca
      301 ttgagtgttt tttatttgtt gtattttttt ttttttagag aaaatcctcc aatatcaaat
      361 taggaatcgt agtttcatga ttttctgtta cacctaactt tttgtgtggt gccctcctcc
      421 ttgtcaatat taatgttaaa gtgcaattct ttttccttat cacgttgagc cattagtatc
      481 aatttgctta cctgtattcc tttactatcc tcctttttct ccttcttgat aaatgtatgt
      541 agattgcgta tatagtttcg tctaccctat gaacatattc cattttgtaa tttcgtgtcg
      601 tttctattat gaatttcatt tataaagttt atgtacaaat atcataaaaa aagagaatct
      661 ttttaagcaa ggattttctt aacttcttcg gcgacagcat caccgacttc ggtggtactg
      721 ttggaaccac ctaaatcacc agttctgata cctgcatcca aaaccttttt aactgcatct
      781 tcaatggcct taccttcttc aggcaagttc aatgacaatt tcaacatcat tgcagcagac
      841 aagatagtgg cgatagggtc aaccttattc tttggcaaat ctggagcaga accgtggcat
      901 ggttcgtaca aaccaaatgc ggtgttcttg tctggcaaag aggccaagga cgcagatggc
      961 aacaaaccca aggaacctgg gataacggag gcttcatcgg agatgatatc accaaacatg
     1021 ttgctggtga ttataatacc atttaggtgg gttgggttct taactaggat catggcggca
     1081 gaatcaatca attgatgttg aaccttcaat gtagggaatt cgttcttgat ggtttcctcc
     1141 acagtttttc tccataatct tgaagaggcc aaaagattag ctttatccaa ggaccaaata
     1201 ggcaatggtg gctcatgttg tagggccatg aaagcggcca ttcttgtgat tctttgcact
     1261 tctggaacgg tgtattgttc actatcccaa gcgacaccat caccatcgtc ttcctttctc
     1321 ttaccaaagt aaatacctcc cactaattct ctgacaacaa cgaagtcagt acctttagca
     1381 aattgtggct tgattggaga taagtctaaa agagagtcgg atgcaaagtt acatggtctt
     1441 aagttggcgt acaattgaag ttctttacgg atttttagta aaccttgttc aggtctaaca
     1501 ctaccggtac cccatttagg accagccaca gcacctaaca aaacggcatc aaccttcttg
     1561 gaggcttcca gcgcctcatc tggaagtggg acacctgtag catcgatagc agcaccacca
     1621 attaaatgat tttcgaaatc gaacttgaca ttggaacgaa catcagaaat agctttaaga
     1681 accttaatgg cttcggctgt gatttcttga ccaacgtggt cacctggcaa aacgacgatc
     1741 ttcttagggg cagacatagg ggcagacatt agaatggtat atccttgaaa tatatatata
     1801 tattgctgaa atgtaaaagg taagaaaagt tagaaagtaa gacgattgct aaccacctat
     1861 tggaaaaaac aataggtcct taaataatat tgtcaacttc aagtattgtg atgcaagcat
     1921 ttagtcatga acgcttctct attctatatg aaaagccggt tccggcctct cacctttcct
     1981 ttttctccca atttttcagt tgaaaaaggt atatgcgtca ggcgacctct gaaattaaca
     2041 aaaaatttcc agtcatcgaa tttgattctg tgcgatagcg cccctgtgtg ttctcgttat
     2101 gttgaggaaa aaaataatgg ttgctaagag attcgaactc ttgcatctta cgatacctga
     2161 gtattcccac agttaactgc ggtcaagata tttcttgaat caggcgcctt agaccgctcg
     2221 gccaaacaac caattacttg ttgagaaata gagtataatt atcctataaa tataacgttt
     2281 ttgaacacac atgaacaagg aagtacagga caattgattt tgaagagaat gtggattttg
     2341 atgtaattgt tgggattcca tttttaataa ggcaataata ttaggtatgt ggatatacta
     2401 gaagttctcc tcgaccgtcg atatgcggtg tgaaataccg cacagatgcg taaggagaaa
     2461 ataccgcatc aggaaattgt aaacgttaat attttgttaa aattcgcgtt aaatttttgt
     2521 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     2581 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     2641 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     2701 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     2761 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     2821 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     2881 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc gcgccattcg
     2941 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
     3001 cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc
     3061 cagtcacgac gttgtaaaac gacggccagt gagcgcgcgt aatacgactc actatagggc
     3121 gaattgggta ccgggccccc cctcgaatca agcttctgaa taagccctcg taatatattt
     3181 tcatgaagaa tttaggtcca aaaaaaagat gggcattaat tctagtcatt taaaaaattc
     3241 tatagatcag aggttacatg gccaagattg aaacttagag gagtatagtt acataaaaga
     3301 aggcaaaacg atgtataaat gaaagaaatt gagatggtgc acgatgcaca gttgaagtga
     3361 acttgcgggg tttttcagta tctacgattc atagatctct cgagctcgaa ttcactagtg
     3421 tcgacctgca gttactcgga tccccttgac gttaaagtat agaggtatat taacaatttt
     3481 ttgttgatac ttttattaca tttgaataag aagtaataca aaccgaaaat gttgaaagta
     3541 ttagttaaag tggttatgca gcttttccat ttatatatct gttaatagat caaaaatcat
     3601 cgcttcgctg attaattacc ccagaaataa ggctaaaaaa ctaatcgcat tatcatccta
     3661 tggttgttaa tttgattcgt taatttgaag gtttgtgggg ccaggttact gccaattttt
     3721 cctcttcata accataaaag ctagtattgt agaatcttta ttgttcggag cagtgcggcg
     3781 cgaggcacat ctgcgtttca ggaacgcgac cggtgaagac gaggacgcac ggaggagagt
     3841 cttccgtcgg agggctgtcg cccgctcggc ggcttctaat ccgtacttca atatagcaat
     3901 gagcagttaa gcgtattact gaaagttcca aagagaaggt ttttttaggc taagataatg
     3961 gggctcttta catttccaca acatataagt aagattagat atggatatgt atatggtggt
     4021 aatgccatgt aatatgatta ttaaacttct ttgcgtccat ccaaaaaaaa agtaagaatt
     4081 tttgaaaatt caatataaat gacagctcag ttacaaagtg aaagccgcgg ctgagtcatt
     4141 ctagtgaccg gcatgcaagc aacgaagcga tggccgccac cgcggtggag ctccagcttt
     4201 tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat catggtcata gctgtttcct
     4261 gtgtgaaatt gttatccgct cacaattcca cacaacatag gagccggaag cataaagtgt
     4321 aaagcctggg gtgcctaatg agtgaggtaa ctcacattaa ttgcgttgcg ctcactgccc
     4381 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     4441 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     4501 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     4561 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     4621 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     4681 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     4741 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     4801 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     4861 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     4921 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     4981 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     5041 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
     5101 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     5161 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     5221 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     5281 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     5341 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     5401 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     5461 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     5521 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     5581 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     5641 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     5701 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     5761 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     5821 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     5881 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     5941 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     6001 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     6061 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     6121 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     6181 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     6241 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     6301 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     6361 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctagttgct tcaggtgctg
     6421 gtgctcgtgg aagctccgca gcagctggcc atgtaggcca acgaagcatc tgtgcttcat
     6481 tttgtagaac aaaaatgcaa cgcgagagcg ctaatttttc aaacaaagaa tctgagctgc
     6541 atttttacag aacagaaatg caacgcgaaa gcgctatttt accaacgaag aatctgtgct
     6601 tcatttttgt aaaacaaaaa tgcaacgcga gagcgctaat ttttcaaaca aagaatctga
     6661 gctgcatttt tacagaacag aaatgcaacg cgagagcgct attttaccaa caaagaatct
     6721 atacttcttt tttgttctac aaaaatgcat cccgagagcg ctatttttct aacaaagcat
     6781 cttagattac tttttttctc ctttgtgcgc tctataatgc agtctcttga taactttttg
     6841 cactgtaggt ccgttaaggt tagaagaagg ctactttggt gtctattttc tcttccataa
     6901 aaaaagcctg actccacttc ccgcgtttac tgattactag cgaagctgcg ggtgcatttt
     6961 ttcaagataa aggcatcccc gattatattc tataccgatg tggattgcgc atactttgtg
     7021 aacagaaagt gatagcgttg atgattcttc attggtcaga aaattatgaa cggtttcttc
     7081 tattttgtct ctatatacta cgtataggaa atgtttacat tttcgtattg ttttcgattc
     7141 actctatgaa tagttcttac tacaattttt ttgtctaaag agtaatacta gagataaaca
     7201 taaaaaatgt agaggtcgag tttagatgca agttcaagga gcgaaaggtg gatgggtagg
     7261 ttatataggg atatagcaca gagatatata gcaaagagat acttttgagc aatgtttgtg
     7321 gaagcggtat tcgcaatatt ttagtagctc gttacagtcc ggtgcgtttt tggttttttg
     7381 aaagtgcgtc ttcagagcgc ttttggtttt caaaagcgct ctgaagttcc tatactttct
     7441 agagaatagg aacttcggaa taggaacttc aaagcgtttc cgaaaacgag cgcttccgaa
     7501 aatgcaacgc gagctgcgca catacagctc actgttcacg tcgcacctat atctgcgtgt
     7561 tgcctgtata tatatataca tgagaagaac ggcatagtgc gtgtttatgc ttaaatgcgt
     7621 acttatatgc gtctatttat gtaggatgaa aggtagtcta gtacctcctg tgatattatc
     7681 ccattccatg cggggtatcg tatgcttcct tcagcactac cctttagctg ttctatatgc
     7741 tgccactcct caattggatt agtctcatcc ttcaatgcta tcatttcctt tgatattgga
     7801 tcataggcca tgtaggccac gaaaaggtct ggatagagaa gttggataat ccaactcttt
     7861 caggacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     7921 aggccctttc gtc
//
