LOCUS       62056_11295        8981 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 8981)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8981
                     /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"
     protein_bind    1287..1411
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        1436..1466
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             1520..2176
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             2521..2823
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(2867..2991)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     terminator      3015..3062
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      3089..3276
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(3300..3316)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3324..3340)
                     /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(3348..3378)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3393..3414)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(3702..4290)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     polyA_signal    complement(4619..4666)
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal (Cole and Stacy, 1985)"
     CDS             complement(4902..5693)
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHDDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     promoter        complement(5694..5798)
                     /label=AmpR promoter
     rep_origin      complement(5825..7167)
                     /direction=LEFT
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     promoter        7426..7707
                     /label=TRP1 promoter
     CDS             7708..8379
                     /codon_start=1
                     /label=TRP1
                     /note="phosphoribosylanthranilate isomerase, required for 
                     tryptophan biosynthesis"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     rep_origin      complement(8485..8940)
                     /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 catatgacaa gtttgtacaa aaaagctgaa cgagaaacgt
     1321 aaaatgatat aaatatcaat atattaaatt agattttgca taaaaaacag actacataat
     1381 actgtaaaac acaacatatc cagtcactat ggcggccgca ttaggcaccc caggctttac
     1441 actttatgct tccggctcgt ataatgtgtg gattttgagt taggatccgt cgagattttc
     1501 aggagctaag gaagctaaaa tggagaaaaa aatcactgga tataccaccg ttgatatatc
     1561 ccaatggcat cgtaaagaac attttgaggc atttcagtca gttgctcaat gtacctataa
     1621 ccagaccgtt cagctggata ttacggcctt tttaaagacc gtaaagaaaa ataagcacaa
     1681 gttttatccg gcctttattc acattcttgc ccgcctgatg aatgctcatc cggaattccg
     1741 tatggcaatg aaagacggtg agctggtgat atgggatagt gttcaccctt gttacaccgt
     1801 tttccatgag caaactgaaa cgttttcatc gctctggagt gaataccacg acgatttccg
     1861 gcagtttcta cacatatatt cgcaagatgt ggcgtgttac ggtgaaaacc tggcctattt
     1921 ccctaaaggg tttattgaga atatgttttt cgtctcagcc aatccctggg tgagtttcac
     1981 cagttttgat ttaaacgtgg ccaatatgga caacttcttc gcccccgttt tcaccatggg
     2041 caaatattat acgcaaggcg acaaggtgct gatgccgctg gcgattcagg ttcatcatgc
     2101 cgtttgtgat ggcttccatg tcggcagaat gcttaatgaa ttacaacagt actgcgatga
     2161 gtggcagggc ggggcgtaaa cgcgtggatc cggcttacta aaagccagat aacagtatgc
     2221 gtatttgcgc gctgattttt gcggtataag aatatatact gatatgtata cccgaagtat
     2281 gtcaaaaaga ggtatgctat gaagcagcgt attacagtga cagttgacag cgacagctat
     2341 cagttgctca aggcatatat gatgtcaata tctccggtct ggtaagcaca accatgcaga
     2401 atgaagcccg tcgtctgcgt gccgaacgct ggaaagcgga aaatcaggaa gggatggctg
     2461 aggtcgcccg gtttattgaa atgaacggct cttttgctga cgagaacagg ggctggtgaa
     2521 atgcagttta aggtttacac ctataaaaga gagagccgtt atcgtctgtt tgtggatgta
     2581 cagagtgata ttattgacac gcccgggcga cggatggtga tccccctggc cagtgcacgt
     2641 ctgctgtcag ataaagtctc ccgtgaactt tacccggtgg tgcatatcgg ggatgaaagc
     2701 tggcgcatga tgaccaccga tatggccagt gtgccggtct ccgttatcgg ggaagaagtg
     2761 gctgatctca gccaccgcga aaatgacatc aaaaacgcca ttaacctgat gttctgggga
     2821 atataaatgt caggctccct tatacacagc cagtctgcag gtcgaccata gtgactggat
     2881 atgttgtgtt ttacagtatt atgtagtctg ttttttatgc aaaatctaat ttaatatatt
     2941 gatatttata tcattttacg tttctcgttc agctttcttg tacaaagtgg tctcgacctg
     3001 cagcggccgc ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt
     3061 tgcgcgcttg cagccaagct aattccgggc gaatttctta tgatttatga tttttattat
     3121 taaataagtt ataaaaaaaa taagtgtata caaattttaa agtgactctt aggttttaaa
     3181 acgaaaattc ttattcttga gtaactcttt cctgtaggtc aggttgcttt ctcaggtata
     3241 gcatgaggtc gctcttattg accacacctc taccggcatg caagcttggc gtaatcatgg
     3301 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     3361 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     3421 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     3481 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     3541 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     3601 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     3661 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     3721 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     3781 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     3841 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     3901 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     3961 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     4021 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     4081 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     4141 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     4201 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     4261 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     4321 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     4381 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     4441 gagtaacctg aggctatggc agggcctgcc gccccgacgt tggctgcgag ccctgggcct
     4501 tcacccgaac ttggggggtg gggtggggaa aaggaagaaa cgcgggcgta ttggccccaa
     4561 tggggtctcg gtggggtatc gacagagtgc cagccctggg accgaacccc gcgtttatga
     4621 acaaacgacc caacaccgtg cgttttattc tgtcttttta ttgccgtcat agcgcgggtt
     4681 ccttccggta ttgtctcctt ccgtgtttca gttagcctcc ccctagggtg ggcgaagaac
     4741 tccagcatga gatccccgcg ctggaggatc atccagccgg cgtcccggaa aacgattccg
     4801 aagcccaacc tttcatagaa ggcggcggtg gaatcgaaat ctcgtgatgg caggttgggc
     4861 gtcgcttggt cggttcattt cgaaccccag agtcccgctc agaagaactc gtcaagaagg
     4921 cgatagaagg cgatgcgctg cgaatcggga gcggcgatac cgtaaagcac gaggaagcgg
     4981 tcagcccatt cgccgccaag ctcttcagca atatcacggg tagccaacgc tatgtcctga
     5041 tagcggtccg ccacacccag ccggccacag tcgatgaatc cagaaaagcg gccattttcc
     5101 accatgatat tcggcaagca ggcatcgtca tgggtcacga cgagatcctc gccgtcgggc
     5161 atgctcgcct tgagcctggc gaacagttcg gctggcgcga gcccctgatg ctcttcgtcc
     5221 agatcatcct gatcgacaag accggcttcc atccgagtac gtgctcgctc gatgcgatgt
     5281 ttcgcttggt ggtcgaatgg gcaggtagcc ggatcaagcg tatgcagccg ccgcattgca
     5341 tcagccatga tggatacttt ctcggcagga gcaaggtgag atgacaggag atcctgcccc
     5401 ggcacttcgc ccaatagcag ccagtccctt cccgcttcag tgacaacgtc gagcacagct
     5461 gcgcaaggaa cgcccgtcgt ggccagccac gatagccgcg ctgcctcgtc ttgcagttca
     5521 ttcagggcac cggacaggtc ggtcttgaca aaaagaaccg ggcgcccctg cgctgacagc
     5581 cggaacacgg cggcatcaga gcagccgatt gtctgttgtg cccagtcata gccgaatagc
     5641 ctctccaccc aagcggccgg agaacctgcg tgcaatccat cttgttcaat catactcttc
     5701 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     5761 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     5821 cctgaacgaa gcatctgtgc ttcattttgt agaacaaaaa tgcaacgcga gagcgctaat
     5881 ttttcaaaca aagaatctga gctgcatttt tacagaacag aaatgcaacg cgaaagcgct
     5941 attttaccaa cgaagaatct gtgcttcatt tttgtaaaac aaaaatgcaa cgcgagagcg
     6001 ctaatttttc aaacaaagaa tctgagctgc atttttacag aacagaaatg caacgcgaga
     6061 gcgctatttt accaacaaag aatctatact tcttttttgt tctacaaaaa tgcatcccga
     6121 gagcgctatt tttctaacaa agcatcttag attacttttt ttctcctttg tgcgctctat
     6181 aatgcagtct cttgataact ttttgcactg taggtccgtt aaggttagaa gaaggctact
     6241 ttggtgtcta ttttctcttc cataaaaaaa gcctgactcc acttcccgcg tttactgatt
     6301 actagcgaag ctgcgggtgc attttttcaa gataaaggca tccccgatta tattctatac
     6361 cgatgtggat tgcgcatact ttgtgaacag aaagtgatag cgttgatgat tcttcattgg
     6421 tcagaaaatt atgaacggtt tcttctattt tgtctctata tactacgtat aggaaatgtt
     6481 tacattttcg tattgttttc gattcactct atgaatagtt cttactacaa tttttttgtc
     6541 taaagagtaa tactagagat aaacataaaa aatgtagagg tcgagtttag atgcaagttc
     6601 aaggagcgaa aggtggatgg gtaggttata tagggatata gcacagagat atatagcaaa
     6661 gagatacttt tgagcaatgt ttgtggaagc ggtattcgca atattttagt agctcgttac
     6721 agtccggtgc gtttttggtt ttttgaaagt gcgtcttcag agcgcttttg gttttcaaaa
     6781 gcgctctgaa gttcctatac tttctagaga ataggaactt cggaatagga acttcaaagc
     6841 gtttccgaaa acgagcgctt ccgaaaatgc aacgcgagct gcgcacatac agctcactgt
     6901 tcacgtcgca cctatatctg cgtgttgcct gtatatatat atacatgaga agaacggcat
     6961 agtgcgtgtt tatgcttaaa tgcgtactta tatgcgtcta tttatgtagg atgaaaggta
     7021 gtctagtacc tcctgtgata ttatcccatt ccatgcgggg tatcgtatgc ttccttcagc
     7081 actacccttt agctgttcta tatgctgcca ctcctcaatt ggattagtct catccttcaa
     7141 tgctatcatt tcctttgata ttggatcata ttaagaaacc attattatca tgacattaac
     7201 ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga
     7261 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     7321 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa
     7381 ctatgcggca tcagagcaga ttgtactgag agtgcaccat agatcaacga cattactata
     7441 tatataatat aggaagcatt taatagaaca gcatcgtaat atatgtgtac tttgcagtta
     7501 tgacgccaga tggcagtagt ggaagatatt ctttattgaa aaatagcttg tcaccttacg
     7561 tacaatcttg atccggagct tttctttttt tgccgattaa gaattaattc ggtcgaaaaa
     7621 agaaaaggag agggccaaga gggagggcat tggtgactat tgagcacgtg agtatacgtg
     7681 attaagcaca caaaggcagc ttggagtatg tctgttatta atttcacagg tagttctggt
     7741 ccattggtga aagtttgcgg cttgcagagc acagaggccg cagaatgtgc tctagattcc
     7801 gatgctgact tgctgggtat tatatgtgtg cccaatagaa agagaacaat tgacccggtt
     7861 attgcaagga aaatttcaag tcttgtaaaa gcatataaaa atagttcagg cactccgaaa
     7921 tacttggttg gcgtgtttcg taatcaacct aaggaggatg ttttggctct ggtcaatgat
     7981 tacggcattg atatcgtcca actgcatgga gatgagtcgt ggcaagaata ccaagagttc
     8041 ctcggtttgc cagttattaa aagactcgta tttccaaaag actgcaacat actactcagt
     8101 gcagcttcac agaaacctca ttcgtttatt cccttgtttg attcagaagc aggtgggaca
     8161 ggtgaacttt tggattggaa ctcgatttct gactgggttg gaaggcaaga gagccccgaa
     8221 agcttacatt ttatgttagc tggtggactg acgccagaaa atgttggtga tgcgcttaga
     8281 ttaaatggcg ttattggtgt tgatgtaagc ggaggtgtgg agacaaatgg tgtaaaagac
     8341 tctaacaaaa tagcaaattt cgtcaaaaat gctaagaaat aggttattac tgagtagtat
     8401 ttatttaagt attgtttgtg cacttgccga tctatgcggt gtgaaatacc gcacagatgc
     8461 gtaaggagaa aataccgcat caggaaattg taaacgttaa tattttgtta aaattcgcgt
     8521 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     8581 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     8641 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     8701 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     8761 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     8821 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     8881 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     8941 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga t
//
