LOCUS       62056_11305        7743 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 7743)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7743
                     /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      1778..1825
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1852..2039
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2063..2079)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2087..2103)
                     /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(2111..2141)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2156..2177)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(2465..3053)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     polyA_signal    complement(3382..3429)
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal (Cole and Stacy, 1985)"
     CDS             complement(3664..4455)
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     promoter        complement(4456..4560)
                     /label=AmpR promoter
     rep_origin      complement(4587..5929)
                     /direction=LEFT
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     promoter        6188..6469
                     /label=TRP1 promoter
     CDS             6470..7141
                     /codon_start=1
                     /label=TRP1
                     /note="phosphoribosylanthranilate isomerase, required for 
                     tryptophan biosynthesis"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     rep_origin      complement(7247..7702)
                     /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 catatgatgt tcagtacaca atcgcaagcc atggattcac
     1321 gaattgcagt agtctgtgtt gttttggcag tgttcgcggt ggaccagacg gtcggagcgc
     1381 cgcaaaaaga tgctgtggca gcaagcggtc ccgcctacac cacaaaatac gaccatattg
     1441 acatcgacca agttttgggt tccaaaagat tagtaaacag ctacgttcaa tgtctgctgg
     1501 acaagaaacc gtgcactccc gaaggagctg aacttagaaa aattttacct gatgccttaa
     1561 aaacccaatg tgtgaaatgc aatgctacgc aaaaaaatgc tgctcttaaa gtagttgacc
     1621 gactccagag agattatgat aaagaatgga aacagcttct tgacaaatgg gaccctaagc
     1681 gtgaatattt ccagaaattc caacaatttt tggcagaaga gaagaaaaaa ggtgttgtca
     1741 aattttaagg atccgtcgac ctgcagcggc cgcataacta gcataacccc ttggggcctc
     1801 taaacgggtc ttgaggggtt ttttgcgcgc ttgcagccaa gctaattccg ggcgaatttc
     1861 ttatgattta tgatttttat tattaaataa gttataaaaa aaataagtgt atacaaattt
     1921 taaagtgact cttaggtttt aaaacgaaaa ttcttattct tgagtaactc tttcctgtag
     1981 gtcaggttgc tttctcaggt atagcatgag gtcgctctta ttgaccacac ctctaccggc
     2041 atgcaagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     2101 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     2161 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     2221 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     2281 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     2341 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     2401 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     2461 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     2521 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     2581 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     2641 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     2701 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     2761 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     2821 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     2881 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     2941 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     3001 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     3061 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     3121 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     3181 ttaaatcaat ctaaagtata tatgagtaac ctgaggctat ggcagggcct gccgccccga
     3241 cgttggctgc gagccctggg ccttcacccg aacttggggg gtggggtggg gaaaaggaag
     3301 aaacgcgggc gtattggccc caatggggtc tcggtggggt atcgacagag tgccagccct
     3361 gggaccgaac cccgcgttta tgaacaaacg acccaacacc gtgcgtttta ttctgtcttt
     3421 ttattgccgt catagcgcgg gttccttccg gtattgtctc cttccgtgtt tcagttagcc
     3481 tccccctagg gtgggcgaag aactccagca tgagatcccc gcgctggagg atcatccagc
     3541 cggcgtcccg gaaaacgatt ccgaagccca acctttcata gaaggcggcg gtggaatcga
     3601 aatctcgtga tggcaggttg ggcgtcgctt ggtcggtcat ttcgaacccc agagtcccgc
     3661 tcagaagaac tcgtcaagaa ggcgatagaa ggcgatgcgc tgcgaatcgg gagcggcgat
     3721 accgtaaagc acgaggaagc ggtcagccca ttcgccgcca agctcttcag caatatcacg
     3781 ggtagccaac gctatgtcct gatagcggtc cgccacaccc agccggccac agtcgatgaa
     3841 tccagaaaag cggccatttt ccaccatgat attcggcaag caggcatcgc catgagtcac
     3901 gacgagatcc tcgccgtcgg gcatgctcgc cttgagcctg gcgaacagtt cggctggcgc
     3961 gagcccctga tgctcttcgt ccagatcatc ctgatcgaca agaccggctt ccatccgagt
     4021 acgtgctcgc tcgatgcgat gtttcgcttg gtggtcgaat gggcaggtag ccggatcaag
     4081 cgtatgcagc cgccgcattg catcagccat gatggatact ttctcggcag gagcaaggtg
     4141 agatgacagg agatcctgcc ccggcacttc gcccaatagc agccagtccc ttcccgcttc
     4201 agtgacaacg tcgagcacag ctgcgcaagg aacgcccgtc gtggccagcc acgatagccg
     4261 cgctgcctcg tcttgcagtt cattcagggc accggacagg tcggtcttga caaaaagaac
     4321 cgggcgcccc tgcgctgaca gccggaacac ggcggcatca gagcagccga ttgtctgttg
     4381 tgcccagtca tagccgaata gcctctccac ccaagcggcc ggagaacctg cgtgcaatcc
     4441 atcttgttca atcatactct tcctttttca atattattga agcatttatc agggttattg
     4501 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     4561 cacatttccc cgaaaagtgc cacctgaacg aagcatctgt gcttcatttt gtagaacaaa
     4621 aatgcaacgc gagagcgcta atttttcaaa caaagaatct gagctgcatt tttacagaac
     4681 agaaatgcaa cgcgaaagcg ctattttacc aacgaagaat ctgtgcttca tttttgtaaa
     4741 acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag aatctgagct gcatttttac
     4801 agaacagaaa tgcaacgcga gagcgctatt ttaccaacaa agaatctata cttctttttt
     4861 gttctacaaa aatgcatccc gagagcgcta tttttctaac aaagcatctt agattacttt
     4921 ttttctcctt tgtgcgctct ataatgcagt ctcttgataa ctttttgcac tgtaggtccg
     4981 ttaaggttag aagaaggcta ctttggtgtc tattttctct tccataaaaa aagcctgact
     5041 ccacttcccg cgtttactga ttactagcga agctgcgggt gcattttttc aagataaagg
     5101 catccccgat tatattctat accgatgtgg attgcgcata ctttgtgaac agaaagtgat
     5161 agcgttgatg attcttcatt ggtcagaaaa ttatgaacgg tttcttctat tttgtctcta
     5221 tatactacgt ataggaaatg tttacatttt cgtattgttt tcgattcact ctatgaatag
     5281 ttcttactac aatttttttg tctaaagagt aatactagag ataaacataa aaaatgtaga
     5341 ggtcgagttt agatgcaagt tcaaggagcg aaaggtggat gggtaggtta tatagggata
     5401 tagcacagag atatatagca aagagatact tttgagcaat gtttgtggaa gcggtattcg
     5461 caatatttta gtagctcgtt acagtccggt gcgtttttgg ttttttgaaa gtgcgtcttc
     5521 agagcgcttt tggttttcaa aagcgctctg aagttcctat actttctaga gaataggaac
     5581 ttcggaatag gaacttcaaa gcgtttccga aaacgagcgc ttccgaaaat gcaacgcgag
     5641 ctgcgcacat acagctcact gttcacgtcg cacctatatc tgcgtgttgc ctgtatatat
     5701 atatacatga gaagaacggc atagtgcgtg tttatgctta aatgcgtact tatatgcgtc
     5761 tatttatgta ggatgaaagg tagtctagta cctcctgtga tattatccca ttccatgcgg
     5821 ggtatcgtat gcttccttca gcactaccct ttagctgttc tatatgctgc cactcctcaa
     5881 ttggattagt ctcatccttc aatgctatca tttcctttga tattggatca tactaagaaa
     5941 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg
     6001 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
     6061 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     6121 gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc
     6181 atagatcaac gacattacta tatatataat ataggaagca tttaatagaa cagcatcgta
     6241 atatatgtgt actttgcagt tatgacgcca gatggcagta gtggaagata ttctttattg
     6301 aaaaatagct tgtcacctta cgtacaatct tgatccggag cttttctttt tttgccgatt
     6361 aagaattaat tcggtcgaaa aaagaaaagg agagggccaa gagggagggc attggtgact
     6421 attgagcacg tgagtatacg tgattaagca cacaaaggca gcttggagta tgtctgttat
     6481 taatttcaca ggtagttctg gtccattggt gaaagtttgc ggcttgcaga gcacagaggc
     6541 cgcagaatgt gctctagatt ccgatgctga cttgctgggt attatatgtg tgcccaatag
     6601 aaagagaaca attgacccgg ttattgcaag gaaaatttca agtcttgtaa aagcatataa
     6661 aaatagttca ggcactccga aatacttggt tggcgtgttt cgtaatcaac ctaaggagga
     6721 tgttttggct ctggtcaatg attacggcat tgatatcgtc caactgcatg gagatgagtc
     6781 gtggcaagaa taccaagagt tcctcggttt gccagttatt aaaagactcg tatttccaaa
     6841 agactgcaac atactactca gtgcagcttc acagaaacct cattcgttta ttcccttgtt
     6901 tgattcagaa gcaggtggga caggtgaact tttggattgg aactcgattt ctgactgggt
     6961 tggaaggcaa gagagccccg aaagcttaca ttttatgtta gctggtggac tgacgccaga
     7021 aaatgttggt gatgcgctta gattaaatgg cgttattggt gttgatgtaa gcggaggtgt
     7081 ggagacaaat ggtgtaaaag actctaacaa aatagcaaat ttcgtcaaaa atgctaagaa
     7141 ataggttatt actgagtagt atttatttaa gtattgtttg tgcacttgcc gatctatgcg
     7201 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggaaat tgtaagcgtt
     7261 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     7321 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     7381 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     7441 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     7501 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     7561 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     7621 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     7681 aatgcgccgc tacagggcgc gtccattcgc cattcaggct gcgcaactgt tgggaagggc
     7741 gat
//
