LOCUS       62056_11300        7804 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 7804)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7804
                     /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      1839..1886
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1913..2100
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2124..2140)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2148..2164)
                     /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(2172..2202)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2217..2238)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(2526..3114)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     polyA_signal    complement(3443..3490)
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal (Cole and Stacy, 1985)"
     CDS             complement(3725..4516)
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     promoter        complement(4517..4621)
                     /label=AmpR promoter
     rep_origin      complement(4648..5990)
                     /direction=LEFT
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     promoter        6249..6530
                     /label=TRP1 promoter
     CDS             6531..7202
                     /codon_start=1
                     /label=TRP1
                     /note="phosphoribosylanthranilate isomerase, required for 
                     tryptophan biosynthesis"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     rep_origin      complement(7308..7763)
                     /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 catatggcca tggaggccga attcccgggg atctctgaag
     1321 aggtggtcag ccgcgaggtg tccggcatca aggccgccta cgaggccgag ctcggggatg
     1381 cccgcaagac ccttgactca gtagccaagg agcgcgcccg cctgcagctg gagctgagca
     1441 aagtgcgtga ggagtttaag gagctgaaag cgcgcaatac caagaaggag ggtgacctga
     1501 tagctgctca ggctcggctg aaggacctgg aggctctgct gaactccaag gaggccgcac
     1561 tgagcactgc tctcagtgag aagcgcacgc tggagggcga gctgcatgat ctgcggggcc
     1621 aggtggccaa gcttgaggca gccctaggtg aggccaagaa gcaacttcag gatgagatgc
     1681 tgcggcgggt ggatgctgag aacaggctgc agaccatgaa ggaggaactg gacttccaga
     1741 agaacatcta cagtgaggag ctgcgtgaga ccaagcgccg tcatgagacc cgactggtgg
     1801 agattgacag gatccgtcga cctgcagcgg ccgcataact agcataaccc cttggggcct
     1861 ctaaacgggt cttgaggggt tttttgcgcg cttgcagcca agctaattcc gggcgaattt
     1921 cttatgattt atgattttta ttattaaata agttataaaa aaaataagtg tatacaaatt
     1981 ttaaagtgac tcttaggttt taaaacgaaa attcttattc ttgagtaact ctttcctgta
     2041 ggtcaggttg ctttctcagg tatagcatga ggtcgctctt attgaccaca cctctaccgg
     2101 catgcaagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     2161 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     2221 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     2281 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     2341 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     2401 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     2461 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     2521 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     2581 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     2641 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     2701 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     2761 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     2821 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     2881 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     2941 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     3001 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     3061 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     3121 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     3181 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     3241 tttaaatcaa tctaaagtat atatgagtaa cctgaggcta tggcagggcc tgccgccccg
     3301 acgttggctg cgagccctgg gccttcaccc gaacttgggg ggtggggtgg ggaaaaggaa
     3361 gaaacgcggg cgtattggcc ccaatggggt ctcggtgggg tatcgacaga gtgccagccc
     3421 tgggaccgaa ccccgcgttt atgaacaaac gacccaacac cgtgcgtttt attctgtctt
     3481 tttattgccg tcatagcgcg ggttccttcc ggtattgtct ccttccgtgt ttcagttagc
     3541 ctccccctag ggtgggcgaa gaactccagc atgagatccc cgcgctggag gatcatccag
     3601 ccggcgtccc ggaaaacgat tccgaagccc aacctttcat agaaggcggc ggtggaatcg
     3661 aaatctcgtg atggcaggtt gggcgtcgct tggtcggtca tttcgaaccc cagagtcccg
     3721 ctcagaagaa ctcgtcaaga aggcgataga aggcgatgcg ctgcgaatcg ggagcggcga
     3781 taccgtaaag cacgaggaag cggtcagccc attcgccgcc aagctcttca gcaatatcac
     3841 gggtagccaa cgctatgtcc tgatagcggt ccgccacacc cagccggcca cagtcgatga
     3901 atccagaaaa gcggccattt tccaccatga tattcggcaa gcaggcatcg ccatgagtca
     3961 cgacgagatc ctcgccgtcg ggcatgctcg ccttgagcct ggcgaacagt tcggctggcg
     4021 cgagcccctg atgctcttcg tccagatcat cctgatcgac aagaccggct tccatccgag
     4081 tacgtgctcg ctcgatgcga tgtttcgctt ggtggtcgaa tgggcaggta gccggatcaa
     4141 gcgtatgcag ccgccgcatt gcatcagcca tgatggatac tttctcggca ggagcaaggt
     4201 gagatgacag gagatcctgc cccggcactt cgcccaatag cagccagtcc cttcccgctt
     4261 cagtgacaac gtcgagcaca gctgcgcaag gaacgcccgt cgtggccagc cacgatagcc
     4321 gcgctgcctc gtcttgcagt tcattcaggg caccggacag gtcggtcttg acaaaaagaa
     4381 ccgggcgccc ctgcgctgac agccggaaca cggcggcatc agagcagccg attgtctgtt
     4441 gtgcccagtc atagccgaat agcctctcca cccaagcggc cggagaacct gcgtgcaatc
     4501 catcttgttc aatcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     4561 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     4621 gcacatttcc ccgaaaagtg ccacctgaac gaagcatctg tgcttcattt tgtagaacaa
     4681 aaatgcaacg cgagagcgct aatttttcaa acaaagaatc tgagctgcat ttttacagaa
     4741 cagaaatgca acgcgaaagc gctattttac caacgaagaa tctgtgcttc atttttgtaa
     4801 aacaaaaatg caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc tgcattttta
     4861 cagaacagaa atgcaacgcg agagcgctat tttaccaaca aagaatctat acttcttttt
     4921 tgttctacaa aaatgcatcc cgagagcgct atttttctaa caaagcatct tagattactt
     4981 tttttctcct ttgtgcgctc tataatgcag tctcttgata actttttgca ctgtaggtcc
     5041 gttaaggtta gaagaaggct actttggtgt ctattttctc ttccataaaa aaagcctgac
     5101 tccacttccc gcgtttactg attactagcg aagctgcggg tgcatttttt caagataaag
     5161 gcatccccga ttatattcta taccgatgtg gattgcgcat actttgtgaa cagaaagtga
     5221 tagcgttgat gattcttcat tggtcagaaa attatgaacg gtttcttcta ttttgtctct
     5281 atatactacg tataggaaat gtttacattt tcgtattgtt ttcgattcac tctatgaata
     5341 gttcttacta caattttttt gtctaaagag taatactaga gataaacata aaaaatgtag
     5401 aggtcgagtt tagatgcaag ttcaaggagc gaaaggtgga tgggtaggtt atatagggat
     5461 atagcacaga gatatatagc aaagagatac ttttgagcaa tgtttgtgga agcggtattc
     5521 gcaatatttt agtagctcgt tacagtccgg tgcgtttttg gttttttgaa agtgcgtctt
     5581 cagagcgctt ttggttttca aaagcgctct gaagttccta tactttctag agaataggaa
     5641 cttcggaata ggaacttcaa agcgtttccg aaaacgagcg cttccgaaaa tgcaacgcga
     5701 gctgcgcaca tacagctcac tgttcacgtc gcacctatat ctgcgtgttg cctgtatata
     5761 tatatacatg agaagaacgg catagtgcgt gtttatgctt aaatgcgtac ttatatgcgt
     5821 ctatttatgt aggatgaaag gtagtctagt acctcctgtg atattatccc attccatgcg
     5881 gggtatcgta tgcttccttc agcactaccc tttagctgtt ctatatgctg ccactcctca
     5941 attggattag tctcatcctt caatgctatc atttcctttg atattggatc atactaagaa
     6001 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc
     6061 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
     6121 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
     6181 ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac
     6241 catagatcaa cgacattact atatatataa tataggaagc atttaataga acagcatcgt
     6301 aatatatgtg tactttgcag ttatgacgcc agatggcagt agtggaagat attctttatt
     6361 gaaaaatagc ttgtcacctt acgtacaatc ttgatccgga gcttttcttt ttttgccgat
     6421 taagaattaa ttcggtcgaa aaaagaaaag gagagggcca agagggaggg cattggtgac
     6481 tattgagcac gtgagtatac gtgattaagc acacaaaggc agcttggagt atgtctgtta
     6541 ttaatttcac aggtagttct ggtccattgg tgaaagtttg cggcttgcag agcacagagg
     6601 ccgcagaatg tgctctagat tccgatgctg acttgctggg tattatatgt gtgcccaata
     6661 gaaagagaac aattgacccg gttattgcaa ggaaaatttc aagtcttgta aaagcatata
     6721 aaaatagttc aggcactccg aaatacttgg ttggcgtgtt tcgtaatcaa cctaaggagg
     6781 atgttttggc tctggtcaat gattacggca ttgatatcgt ccaactgcat ggagatgagt
     6841 cgtggcaaga ataccaagag ttcctcggtt tgccagttat taaaagactc gtatttccaa
     6901 aagactgcaa catactactc agtgcagctt cacagaaacc tcattcgttt attcccttgt
     6961 ttgattcaga agcaggtggg acaggtgaac ttttggattg gaactcgatt tctgactggg
     7021 ttggaaggca agagagcccc gaaagcttac attttatgtt agctggtgga ctgacgccag
     7081 aaaatgttgg tgatgcgctt agattaaatg gcgttattgg tgttgatgta agcggaggtg
     7141 tggagacaaa tggtgtaaaa gactctaaca aaatagcaaa tttcgtcaaa aatgctaaga
     7201 aataggttat tactgagtag tatttattta agtattgttt gtgcacttgc cgatctatgc
     7261 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggaaa ttgtaagcgt
     7321 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     7381 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
     7441 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     7501 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
     7561 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     7621 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     7681 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     7741 taatgcgccg ctacagggcg cgtccattcg ccattcaggc tgcgcaactg ttgggaaggg
     7801 cgat
//
