LOCUS       62056_11310        7749 bp DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7749)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7749
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        30..734
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             762..1202
                     /codon_start=1
                     /label=GAL4 DNA binding domain
                     /note="DNA binding domain of the GAL4 transcriptional
                     activator"
                     /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK
                     RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK
                     DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"
     promoter        1213..1231
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1251..1280
                     /codon_start=1
                     /label=Myc
                     /note="Myc (human c-Myc proto-oncogene) epitope tag"
                     /translation="EQKLISEEDL"
     terminator      1784..1831
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1858..2045
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2069..2085)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2093..2109)
                     /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(2117..2147)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2162..2183)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(2471..3059)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     polyA_signal    complement(3388..3435)
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal (Cole and Stacy, 1985)"
     CDS             complement(3670..4461)
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     promoter        complement(4462..4566)
                     /label=AmpR promoter
     rep_origin      complement(4593..5935)
                     /direction=LEFT
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     promoter        6194..6475
                     /label=TRP1 promoter
     CDS             6476..7147
                     /codon_start=1
                     /label=TRP1
                     /note="phosphoribosylanthranilate isomerase, required for 
                     tryptophan biosynthesis"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     rep_origin      complement(7253..7708)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 cggtgcgggc ctcttcgcta ttacgccaga tccttttgtt gtttccgggt gtacaatatg
       61 gacttcctct tttctggcaa ccaaacccat acatcgggat tcctataata ccttcgttgg
      121 tctccctaac atgtaggtgg cggaggggag atatacaata gaacagatac cagacaagac
      181 ataatgggct aaacaagact acaccaatta cactgcctca ttgatggtgg tacataacga
      241 actaatactg tagccctaga cttgatagcc atcatcatat cgaagtttca ctaccctttt
      301 tccatttgcc atctattgaa gtaataatag gcgcatgcaa cttcttttct ttttttttct
      361 tttctctctc ccccgttgtt gtctcaccat atccgcaatg acaaaaaaat gatggaagac
      421 actaaaggaa aaaattaacg acaaagacag caccaacaga tgtcgttgtt ccagagctga
      481 tgaggggtat ctcgaagcac acgaaacttt ttccttcctt cattcacgca cactactctc
      541 taatgagcaa cggtatacgg ccttccttcc agttacttga atttgaaata aaaaaaagtt
      601 tgctgtcttg ctatcaagta taaatagacc tgcaattatt aatcttttgt ttcctcgtca
      661 ttgttctcgt tccctttctt ccttgtttct ttttctgcac aatatttcaa gctataccaa
      721 gcatacaatc aactccaagc ttgaagcaag cctcctgaaa gatgaagcta ctgtcttcta
      781 tcgaacaagc atgcgatatt tgccgactta aaaagctcaa gtgctccaaa gaaaaaccga
      841 agtgcgccaa gtgtctgaag aacaactggg agtgtcgcta ctctcccaaa accaaaaggt
      901 ctccgctgac tagggcacat ctgacagaag tggaatcaag gctagaaaga ctggaacagc
      961 tatttctact gatttttcct cgagaagacc ttgacatgat tttgaaaatg gattctttac
     1021 aggatataaa agcattgtta acaggattat ttgtacaaga taatgtgaat aaagatgccg
     1081 tcacagatag attggcttca gtggagactg atatgcctct aacattgaga cagcatagaa
     1141 taagtgcgac atcatcatcg gaagagagta gtaacaaagg tcaaagacag ttgactgtat
     1201 cgccggaatt tgtaatacga ctcactatag ggcgagccgc catcatggag gagcagaagc
     1261 tgatctcaga ggaggacctg catatgatgg ctttcaagga aatcgcaatg ttttccagtt
     1321 tagtggtggt catcataagc tacaatgtac tagaaagcaa tgcattaatg atgcaaccat
     1381 taatagacca agatttctgt gcagtaaaag ctagtttgta cgaattggga gaggctgaga
     1441 tgaacttgat ggacgattcg cagacattaa acgacatgca aagagaatat tttgctggaa
     1501 aagttgacta ttctgctgtg gaaagagcaa ggaatgaact taataagtcc aaaaacaagt
     1561 ggttccaaaa agttatacaa tatctcaagg caacaaatga atttgagccc acaatcaact
     1621 atcaaacctc agatccacaa aaagtttacc atcaaatgga tgacttggac aattataaag
     1681 atgctctaga tgatatgcat gaaaatatcc ttaaatcaat gtcccctacc gtagcccaac
     1741 ctgttgtagc ataaggatcc gtcgacctgc agcggccgca taactagcat aaccccttgg
     1801 ggcctctaaa cgggtcttga ggggtttttt gcgcgcttgc agccaagcta attccgggcg
     1861 aatttcttat gatttatgat ttttattatt aaataagtta taaaaaaaat aagtgtatac
     1921 aaattttaaa gtgactctta ggttttaaaa cgaaaattct tattcttgag taactctttc
     1981 ctgtaggtca ggttgctttc tcaggtatag catgaggtcg ctcttattga ccacacctct
     2041 accggcatgc aagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     2101 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     2161 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     2221 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     2281 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     2341 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     2401 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     2461 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     2521 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     2581 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     2641 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     2701 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     2761 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     2821 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     2881 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     2941 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     3001 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     3061 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     3121 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     3181 gaagttttaa atcaatctaa agtatatatg agtaacctga ggctatggca gggcctgccg
     3241 ccccgacgtt ggctgcgagc cctgggcctt cacccgaact tggggggtgg ggtggggaaa
     3301 aggaagaaac gcgggcgtat tggccccaat ggggtctcgg tggggtatcg acagagtgcc
     3361 agccctggga ccgaaccccg cgtttatgaa caaacgaccc aacaccgtgc gttttattct
     3421 gtctttttat tgccgtcata gcgcgggttc cttccggtat tgtctccttc cgtgtttcag
     3481 ttagcctccc cctagggtgg gcgaagaact ccagcatgag atccccgcgc tggaggatca
     3541 tccagccggc gtcccggaaa acgattccga agcccaacct ttcatagaag gcggcggtgg
     3601 aatcgaaatc tcgtgatggc aggttgggcg tcgcttggtc ggtcatttcg aaccccagag
     3661 tcccgctcag aagaactcgt caagaaggcg atagaaggcg atgcgctgcg aatcgggagc
     3721 ggcgataccg taaagcacga ggaagcggtc agcccattcg ccgccaagct cttcagcaat
     3781 atcacgggta gccaacgcta tgtcctgata gcggtccgcc acacccagcc ggccacagtc
     3841 gatgaatcca gaaaagcggc cattttccac catgatattc ggcaagcagg catcgccatg
     3901 agtcacgacg agatcctcgc cgtcgggcat gctcgccttg agcctggcga acagttcggc
     3961 tggcgcgagc ccctgatgct cttcgtccag atcatcctga tcgacaagac cggcttccat
     4021 ccgagtacgt gctcgctcga tgcgatgttt cgcttggtgg tcgaatgggc aggtagccgg
     4081 atcaagcgta tgcagccgcc gcattgcatc agccatgatg gatactttct cggcaggagc
     4141 aaggtgagat gacaggagat cctgccccgg cacttcgccc aatagcagcc agtcccttcc
     4201 cgcttcagtg acaacgtcga gcacagctgc gcaaggaacg cccgtcgtgg ccagccacga
     4261 tagccgcgct gcctcgtctt gcagttcatt cagggcaccg gacaggtcgg tcttgacaaa
     4321 aagaaccggg cgcccctgcg ctgacagccg gaacacggcg gcatcagagc agccgattgt
     4381 ctgttgtgcc cagtcatagc cgaatagcct ctccacccaa gcggccggag aacctgcgtg
     4441 caatccatct tgttcaatca tactcttcct ttttcaatat tattgaagca tttatcaggg
     4501 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     4561 tccgcgcaca tttccccgaa aagtgccacc tgaacgaagc atctgtgctt cattttgtag
     4621 aacaaaaatg caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc tgcattttta
     4681 cagaacagaa atgcaacgcg aaagcgctat tttaccaacg aagaatctgt gcttcatttt
     4741 tgtaaaacaa aaatgcaacg cgagagcgct aatttttcaa acaaagaatc tgagctgcat
     4801 ttttacagaa cagaaatgca acgcgagagc gctattttac caacaaagaa tctatacttc
     4861 ttttttgttc tacaaaaatg catcccgaga gcgctatttt tctaacaaag catcttagat
     4921 tacttttttt ctcctttgtg cgctctataa tgcagtctct tgataacttt ttgcactgta
     4981 ggtccgttaa ggttagaaga aggctacttt ggtgtctatt ttctcttcca taaaaaaagc
     5041 ctgactccac ttcccgcgtt tactgattac tagcgaagct gcgggtgcat tttttcaaga
     5101 taaaggcatc cccgattata ttctataccg atgtggattg cgcatacttt gtgaacagaa
     5161 agtgatagcg ttgatgattc ttcattggtc agaaaattat gaacggtttc ttctattttg
     5221 tctctatata ctacgtatag gaaatgttta cattttcgta ttgttttcga ttcactctat
     5281 gaatagttct tactacaatt tttttgtcta aagagtaata ctagagataa acataaaaaa
     5341 tgtagaggtc gagtttagat gcaagttcaa ggagcgaaag gtggatgggt aggttatata
     5401 gggatatagc acagagatat atagcaaaga gatacttttg agcaatgttt gtggaagcgg
     5461 tattcgcaat attttagtag ctcgttacag tccggtgcgt ttttggtttt ttgaaagtgc
     5521 gtcttcagag cgcttttggt tttcaaaagc gctctgaagt tcctatactt tctagagaat
     5581 aggaacttcg gaataggaac ttcaaagcgt ttccgaaaac gagcgcttcc gaaaatgcaa
     5641 cgcgagctgc gcacatacag ctcactgttc acgtcgcacc tatatctgcg tgttgcctgt
     5701 atatatatat acatgagaag aacggcatag tgcgtgttta tgcttaaatg cgtacttata
     5761 tgcgtctatt tatgtaggat gaaaggtagt ctagtacctc ctgtgatatt atcccattcc
     5821 atgcggggta tcgtatgctt ccttcagcac taccctttag ctgttctata tgctgccact
     5881 cctcaattgg attagtctca tccttcaatg ctatcatttc ctttgatatt ggatcatact
     5941 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     6001 gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg
     6061 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
     6121 gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag
     6181 tgcaccatag atcaacgaca ttactatata tataatatag gaagcattta atagaacagc
     6241 atcgtaatat atgtgtactt tgcagttatg acgccagatg gcagtagtgg aagatattct
     6301 ttattgaaaa atagcttgtc accttacgta caatcttgat ccggagcttt tctttttttg
     6361 ccgattaaga attaattcgg tcgaaaaaag aaaaggagag ggccaagagg gagggcattg
     6421 gtgactattg agcacgtgag tatacgtgat taagcacaca aaggcagctt ggagtatgtc
     6481 tgttattaat ttcacaggta gttctggtcc attggtgaaa gtttgcggct tgcagagcac
     6541 agaggccgca gaatgtgctc tagattccga tgctgacttg ctgggtatta tatgtgtgcc
     6601 caatagaaag agaacaattg acccggttat tgcaaggaaa atttcaagtc ttgtaaaagc
     6661 atataaaaat agttcaggca ctccgaaata cttggttggc gtgtttcgta atcaacctaa
     6721 ggaggatgtt ttggctctgg tcaatgatta cggcattgat atcgtccaac tgcatggaga
     6781 tgagtcgtgg caagaatacc aagagttcct cggtttgcca gttattaaaa gactcgtatt
     6841 tccaaaagac tgcaacatac tactcagtgc agcttcacag aaacctcatt cgtttattcc
     6901 cttgtttgat tcagaagcag gtgggacagg tgaacttttg gattggaact cgatttctga
     6961 ctgggttgga aggcaagaga gccccgaaag cttacatttt atgttagctg gtggactgac
     7021 gccagaaaat gttggtgatg cgcttagatt aaatggcgtt attggtgttg atgtaagcgg
     7081 aggtgtggag acaaatggtg taaaagactc taacaaaata gcaaatttcg tcaaaaatgc
     7141 taagaaatag gttattactg agtagtattt atttaagtat tgtttgtgca cttgccgatc
     7201 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggaaattgta
     7261 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     7321 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     7381 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     7441 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     7501 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     7561 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     7621 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     7681 gcgcttaatg cgccgctaca gggcgcgtcc attcgccatt caggctgcgc aactgttggg
     7741 aagggcgat
//
