LOCUS       62056_11290        8275 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 8275)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8275
                     /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"
     CDS             complement(1464..1475)
                     /codon_start=1
                     /label=Factor Xa site
                     /note="Factor Xa recognition and cleavage site"
                     /translation="IEGR"
     terminator      2309..2356
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2383..2570
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2594..2610)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2618..2634)
                     /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(2642..2672)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2687..2708)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(2996..3584)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     polyA_signal    complement(3913..3960)
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal (Cole and Stacy, 1985)"
     CDS             complement(4196..4987)
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHDDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     promoter        complement(4988..5092)
                     /label=AmpR promoter
     rep_origin      complement(5119..6461)
                     /direction=LEFT
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     promoter        6720..7001
                     /label=TRP1 promoter
     CDS             7002..7673
                     /codon_start=1
                     /label=TRP1
                     /note="phosphoribosylanthranilate isomerase, required for 
                     tryptophan biosynthesis"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     rep_origin      complement(7779..8234)
                     /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 acaaaaaaaa tgatggaaga
      421 cactaaagga aaaaattaac gacaaagaca gcaccaacag atgtcgttgt tccagagctg
      481 atgaggggta tctcgaagca cacgaaactt tttccttcct tcattcacgc acactactct
      541 ctaatgagca acggtatacg gccttccttc cagttacttg aatttgaaat 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 attccctgtc accgagaccc
     1321 ctgggccagt ggcccctgcc ccagccactc catggcccct gtcatctttt gtcccttctc
     1381 aaaaaactta ccagggcaac tatggcttcc acctgggctt cctgcagtct gggacagcca
     1441 agtctgttat gtgcacgtac tctcctcccc tcaataagct attctgccag ctggcgaaga
     1501 cgtgccctgt gcagttgtgg gtcagcgcca cacctccagc tgggagccgt gtccgcgcca
     1561 tggccatcta caagaagtca cagcacatga cggaggtcgt gagacgctgc ccccaccatg
     1621 agcgctgctc cgatggtgat ggcctggctc ctccccagca tcttatccgg gtggaaggaa
     1681 atttgtatcc cgagtatctg gaagacaggc agacttttcg ccacagcgtg gtggtacctt
     1741 atgagccacc cgaggccggc tctgagtata ccaccatcca ctacaagtac atgtgtaata
     1801 gctcctgcat ggggggcatg aaccgccgac ctatccttac catcatcaca ctggaagact
     1861 ccagtgggaa ccttctggga cgggacagct ttgaggttcg tgtttgtgcc tgccctggga
     1921 gagaccgccg tacagaagaa gaaaatttcc gcaaaaagga agtcctttgc cctgaactgc
     1981 ccccagggag cgcaaagaga gcgctgccca cctgcacaag cgcctctccc ccgcaaaaga
     2041 aaaaaccact tgatggagag tatttcaccc tcaagatccg cgggcgtaaa cgcttcgaga
     2101 tgttccggga gctgaatgag gccttagagt taaaggatgc ccatgctaca gaggagtctg
     2161 gagacagcag ggctcactcc agctacctga agaccaagaa gggccagtct acttcccgcc
     2221 ataaaaaaac aatggtcaag aaagtggggc ctgactcaga ctgactgcct ctgcatcccg
     2281 gatccgtcga cctgcagcgg ccgcataact agcataaccc cttggggcct ctaaacgggt
     2341 cttgaggggt tttttgcgcg cttgcagcca agctaattcc gggcgaattt cttatgattt
     2401 atgattttta ttattaaata agttataaaa aaaataagtg tatacaaatt ttaaagtgac
     2461 tcttaggttt taaaacgaaa attcttattc ttgagtaact ctttcctgta ggtcaggttg
     2521 ctttctcagg tatagcatga ggtcgctctt attgaccaca cctctaccgg catgcaagct
     2581 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     2641 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     2701 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     2761 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     2821 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     2881 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     2941 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     3001 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     3061 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     3121 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     3181 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     3241 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     3301 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     3361 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     3421 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     3481 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     3541 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     3601 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     3661 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     3721 tctaaagtat atatgagtaa cctgaggcta tggcagggcc tgccgccccg acgttggctg
     3781 cgagccctgg gccttcaccc gaacttgggg ggtggggtgg ggaaaaggaa gaaacgcggg
     3841 cgtattggcc ccaatggggt ctcggtgggg tatcgacaga gtgccagccc tgggaccgaa
     3901 ccccgcgttt atgaacaaac gacccaacac cgtgcgtttt attctgtctt tttattgccg
     3961 tcatagcgcg ggttccttcc ggtattgtct ccttccgtgt ttcagttagc ctccccctag
     4021 ggtgggcgaa gaactccagc atgagatccc cgcgctggag gatcatccag ccggcgtccc
     4081 ggaaaacgat tccgaagccc aacctttcat agaaggcggc ggtggaatcg aaatctcgtg
     4141 atggcaggtt gggcgtcgct tggtcggttc atttcgaacc ccagagtccc gctcagaaga
     4201 actcgtcaag aaggcgatag aaggcgatgc gctgcgaatc gggagcggcg ataccgtaaa
     4261 gcacgaggaa gcggtcagcc cattcgccgc caagctcttc agcaatatca cgggtagcca
     4321 acgctatgtc ctgatagcgg tccgccacac ccagccggcc acagtcgatg aatccagaaa
     4381 agcggccatt ttccaccatg atattcggca agcaggcatc gtcatgggtc acgacgagat
     4441 cctcgccgtc gggcatgctc gccttgagcc tggcgaacag ttcggctggc gcgagcccct
     4501 gatgctcttc gtccagatca tcctgatcga caagaccggc ttccatccga gtacgtgctc
     4561 gctcgatgcg atgtttcgct tggtggtcga atgggcaggt agccggatca agcgtatgca
     4621 gccgccgcat tgcatcagcc atgatggata ctttctcggc aggagcaagg tgagatgaca
     4681 ggagatcctg ccccggcact tcgcccaata gcagccagtc ccttcccgct tcagtgacaa
     4741 cgtcgagcac agctgcgcaa ggaacgcccg tcgtggccag ccacgatagc cgcgctgcct
     4801 cgtcttgcag ttcattcagg gcaccggaca ggtcggtctt gacaaaaaga accgggcgcc
     4861 cctgcgctga cagccggaac acggcggcat cagagcagcc gattgtctgt tgtgcccagt
     4921 catagccgaa tagcctctcc acccaagcgg ccggagaacc tgcgtgcaat ccatcttgtt
     4981 caatcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     5041 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     5101 cccgaaaagt gccacctgaa cgaagcatct gtgcttcatt ttgtagaaca aaaatgcaac
     5161 gcgagagcgc taatttttca aacaaagaat ctgagctgca tttttacaga acagaaatgc
     5221 aacgcgaaag cgctatttta ccaacgaaga atctgtgctt catttttgta aaacaaaaat
     5281 gcaacgcgag agcgctaatt tttcaaacaa agaatctgag ctgcattttt acagaacaga
     5341 aatgcaacgc gagagcgcta ttttaccaac aaagaatcta tacttctttt ttgttctaca
     5401 aaaatgcatc ccgagagcgc tatttttcta acaaagcatc ttagattact ttttttctcc
     5461 tttgtgcgct ctataatgca gtctcttgat aactttttgc actgtaggtc cgttaaggtt
     5521 agaagaaggc tactttggtg tctattttct cttccataaa aaaagcctga ctccacttcc
     5581 cgcgtttact gattactagc gaagctgcgg gtgcattttt tcaagataaa ggcatccccg
     5641 attatattct ataccgatgt ggattgcgca tactttgtga acagaaagtg atagcgttga
     5701 tgattcttca ttggtcagaa aattatgaac ggtttcttct attttgtctc tatatactac
     5761 gtataggaaa tgtttacatt ttcgtattgt tttcgattca ctctatgaat agttcttact
     5821 acaatttttt tgtctaaaga gtaatactag agataaacat aaaaaatgta gaggtcgagt
     5881 ttagatgcaa gttcaaggag cgaaaggtgg atgggtaggt tatataggga tatagcacag
     5941 agatatatag caaagagata cttttgagca atgtttgtgg aagcggtatt cgcaatattt
     6001 tagtagctcg ttacagtccg gtgcgttttt ggttttttga aagtgcgtct tcagagcgct
     6061 tttggttttc aaaagcgctc tgaagttcct atactttcta gagaatagga acttcggaat
     6121 aggaacttca aagcgtttcc gaaaacgagc gcttccgaaa atgcaacgcg agctgcgcac
     6181 atacagctca ctgttcacgt cgcacctata tctgcgtgtt gcctgtatat atatatacat
     6241 gagaagaacg gcatagtgcg tgtttatgct taaatgcgta cttatatgcg tctatttatg
     6301 taggatgaaa ggtagtctag tacctcctgt gatattatcc cattccatgc ggggtatcgt
     6361 atgcttcctt cagcactacc ctttagctgt tctatatgct gccactcctc aattggatta
     6421 gtctcatcct tcaatgctat catttccttt gatattggat catattaaga aaccattatt
     6481 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc
     6541 ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg
     6601 taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt
     6661 cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatagatca
     6721 acgacattac tatatatata atataggaag catttaatag aacagcatcg taatatatgt
     6781 gtactttgca gttatgacgc cagatggcag tagtggaaga tattctttat tgaaaaatag
     6841 cttgtcacct tacgtacaat cttgatccgg agcttttctt tttttgccga ttaagaatta
     6901 attcggtcga aaaaagaaaa ggagagggcc aagagggagg gcattggtga ctattgagca
     6961 cgtgagtata cgtgattaag cacacaaagg cagcttggag tatgtctgtt attaatttca
     7021 caggtagttc tggtccattg gtgaaagttt gcggcttgca gagcacagag gccgcagaat
     7081 gtgctctaga ttccgatgct gacttgctgg gtattatatg tgtgcccaat agaaagagaa
     7141 caattgaccc ggttattgca aggaaaattt caagtcttgt aaaagcatat aaaaatagtt
     7201 caggcactcc gaaatacttg gttggcgtgt ttcgtaatca acctaaggag gatgttttgg
     7261 ctctggtcaa tgattacggc attgatatcg tccaactgca tggagatgag tcgtggcaag
     7321 aataccaaga gttcctcggt ttgccagtta ttaaaagact cgtatttcca aaagactgca
     7381 acatactact cagtgcagct tcacagaaac ctcattcgtt tattcccttg tttgattcag
     7441 aagcaggtgg gacaggtgaa cttttggatt ggaactcgat ttctgactgg gttggaaggc
     7501 aagagagccc cgaaagctta cattttatgt tagctggtgg actgacgcca gaaaatgttg
     7561 gtgatgcgct tagattaaat ggcgttattg gtgttgatgt aagcggaggt gtggagacaa
     7621 atggtgtaaa agactctaac aaaatagcaa atttcgtcaa aaatgctaag aaataggtta
     7681 ttactgagta gtatttattt aagtattgtt tgtgcacttg ccgatctatg cggtgtgaaa
     7741 taccgcacag atgcgtaagg agaaaatacc gcatcaggaa attgtaaacg ttaatatttt
     7801 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     7861 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     7921 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     7981 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     8041 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     8101 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     8161 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     8221 gctacagggc gcgtccattc gccattcagg ctgcgcaact gttgggaagg gcgat
//
