LOCUS       Exported                7495 bp ds-DNA     circular SYN 15-DEC-2017
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pBSWHhc-ccdB
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 7495)
  AUTHORS   Mireia
  TITLE     Direct Submission
  JOURNAL   Exported 20 Aug 2019 from SnapGene 2.6.2
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7495
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     promoter        30..734
                     /gene="S. cerevisiae ADH1"
                     /note="ADH1 promoter"
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             762..1202
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="DNA binding domain of the GAL4 transcriptional 
                     activator"
                     /note="GAL4 DNA binding domain"
                     /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK
                     RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK
                     DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"
     promoter        1213..1231
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1245..1247
                     /codon_start=1
                     /product="start codon for in vitro 
                     transcription/translation"
                     /note="ATG"
                     /translation="M"
     CDS             1251..1280
                     /codon_start=1
                     /product="Myc (human c-Myc oncogene) epitope tag"
                     /note="Myc"
                     /translation="EQKLISEEDL"
     misc_feature    1281..1309
                     /label=MCS
                     /note="MCS"
                     /note="multiple cloning site"
     CDS             1310
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="activation domain of the GAL4 transcriptional 
                     activator"
                     /note="GAL4 activation domain"
                     /translation=""
     protein_bind    1317..1441
                     /gene="mutant version of attR"
                     /bound_moiety="LR Clonase(TM)"
                     /note="attR1"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        1466..1496
                     /note="lac UV5 promoter"
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             1550..2209
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /note="CmR"
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             2551..2856
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /note="ccdB"
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(2897..3021)
                     /gene="mutant version of attR"
                     /bound_moiety="LR Clonase(TM)"
                     /note="attR2"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    3032..3055
                     /label=MCS
                     /note="MCS"
                     /note="multiple cloning site"
     terminator      3060..3107
                     /note="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      3134..3228
                     /gene="S. cerevisiae ADH1"
                     /note="ADH1 terminator"
                     /note="transcription terminator for alcohol dehydrogenase 
                     1"
     misc_feature    3229..3288
                     /note="Homology region 2"
     misc_feature    3285..3335
                     /note="Self_overlap_site"
     misc_feature    3336..3357
                     /note="Primer"
     misc_feature    3470..3496
                     /note="primer"
     rep_origin      complement(3650..4238)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number colE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    4567..4614
                     /note="HSV TK poly(A) signal"
                     /note="herpesvirus thymidine kinase polyadenylation signal"
     CDS             complement(4846..5637)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /note="SmR"
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     CDS             complement(4846)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /note="NeoR/KanR"
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin)"
                     /translation=""
     CDS             complement(4846)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /note="NeoR/KanR"
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin)"
                     /translation=""
     CDS             complement(5634..5637)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /note="NeoR/KanR"
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin)"
                     /translation="M"
     promoter        complement(5638..5742)
                     /gene="bla"
                     /note="AmpR promoter"
     misc_feature    5795..5817
                     /note="primer"
     misc_feature    5818..5868
                     /note="Self_overlap_site"
     misc_feature    5865..5924
                     /note="Homology region 1"
     promoter        5940..6221
                     /gene="S. cerevisiae TRP1"
                     /note="TRP1 promoter"
     CDS             6222..6896
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /note="TRP1"
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     rep_origin      complement(6999..7454)
                     /direction=LEFT
                     /note="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 atctccacaa
     1321 gtttgtacaa aaaagctgaa cgagaaacgt aaaatgatat aaatatcaat atattaaatt
     1381 agattttgca taaaaaacag actacataat actgtaaaac acaacatatc cagtcactat
     1441 ggcggccgca ttaggcaccc caggctttac actttatgct tccggctcgt ataatgtgtg
     1501 gattttgagt taggatccgt cgagattttc aggagctaag gaagctaaaa tggagaaaaa
     1561 aatcactgga tataccaccg ttgatatatc ccaatggcat cgtaaagaac attttgaggc
     1621 atttcagtca gttgctcaat gtacctataa ccagaccgtt cagctggata ttacggcctt
     1681 tttaaagacc gtaaagaaaa ataagcacaa gttttatccg gcctttattc acattcttgc
     1741 ccgcctgatg aatgctcatc cggaattccg tatggcaatg aaagacggtg agctggtgat
     1801 atgggatagt gttcaccctt gttacaccgt tttccatgag caaactgaaa cgttttcatc
     1861 gctctggagt gaataccacg acgatttccg gcagtttcta cacatatatt cgcaagatgt
     1921 ggcgtgttac ggtgaaaacc tggcctattt ccctaaaggg tttattgaga atatgttttt
     1981 cgtctcagcc aatccctggg tgagtttcac cagttttgat ttaaacgtgg ccaatatgga
     2041 caacttcttc gcccccgttt tcaccatggg caaatattat acgcaaggcg acaaggtgct
     2101 gatgccgctg gcgattcagg ttcatcatgc cgtttgtgat ggcttccatg tcggcagaat
     2161 gcttaatgaa ttacaacagt actgcgatga gtggcagggc ggggcgtaaa cgcgtggatc
     2221 cggcttacta aaagccagat aacagtatgc gtatttgcgc gctgattttt gcggtataag
     2281 aatatatact gatatgtata cccgaagtat gtcaaaaaga ggtatgctat gaagcagcgt
     2341 attacagtga cagttgacag cgacagctat cagttgctca aggcatatat gatgtcaata
     2401 tctccggtct ggtaagcaca accatgcaga atgaagcccg tcgtctgcgt gccgaacgct
     2461 ggaaagcgga aaatcaggaa gggatggctg aggtcgcccg gtttattgaa atgaacggct
     2521 cttttgctga cgagaacagg ggctggtgaa atgcagttta aggtttacac ctataaaaga
     2581 gagagccgtt atcgtctgtt tgtggatgta cagagtgata ttattgacac gcccgggcga
     2641 cggatggtga tccccctggc cagtgcacgt ctgctgtcag ataaagtctc ccgtgaactt
     2701 tacccggtgg tgcatatcgg ggatgaaagc tggcgcatga tgaccaccga tatggccagt
     2761 gtgccggtct ccgttatcgg ggaagaagtg gctgatctca gccaccgcga aaatgacatc
     2821 aaaaacgcca ttaacctgat gttctgggga atataaatgt caggctccct tatacacagc
     2881 cagtctgcag gtcgaccata gtgactggat atgttgtgtt ttacagtatt atgtagtctg
     2941 ttttttatgc aaaatctaat ttaatatatt gatatttata tcattttacg tttctcgttc
     3001 agctttcttg tacaaagtgg taagtaacta agatccgtcg acctgcagcg gccgcataac
     3061 tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgcgc gcttgcagcc
     3121 aagctaattc cgggcgaatt tcttatgatt tatgattttt attattaaat aagttataaa
     3181 aaaaataagt gtatacaaat tttaaagtga ctcttaggtt ttaaaacgtt atccctagtt
     3241 taaacaccca taatacccat aatagctgtt tgccaattac cctgttatcc ctattacttc
     3301 taaatctgag gaggaaaatc gaatcgctac cttagcgcct gtttcaggct gtctatgtgt
     3361 gactgttgag ctgtaacaag ttgtctcagg tgttcaattt catgttctag ttgctttgtt
     3421 ttactggttt cacctgttct attaggtgtt acatgctgtt catctgttac attgtcgatc
     3481 tgttcatggt gaacaggcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     3541 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     3601 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     3661 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     3721 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     3781 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     3841 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     3901 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     3961 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     4021 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     4081 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     4141 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     4201 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     4261 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     4321 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     4381 gtatatatga gtaacctgag gctatggcag ggcctgccgc cccgacgttg gctgcgagcc
     4441 ctgggccttc acccgaactt ggggggtggg gtggggaaaa ggaagaaacg cgggcgtatt
     4501 ggccccaatg gggtctcggt ggggtatcga cagagtgcca gccctgggac cgaaccccgc
     4561 gtttatgaac aaacgaccca acaccgtgcg ttttattctg tctttttatt gccgtcatag
     4621 cgcgggttcc ttccggtatt gtctccttcc gtgtttcagt tagcctcccc ctagggtggg
     4681 cgaagaactc cagcatgaga tccccgcgct ggaggatcat ccagccggcg tcccggaaaa
     4741 cgattccgaa gcccaacctt tcatagaagg cggcggtgga atcgaaatct cgtgatggca
     4801 ggttgggcgt cgcttggtcg gtcatttcga accccagagt cccgcttatt tgccgactac
     4861 cttggtgatc tcgcctttca cgtagtggac aaattcttcc aactgatctg cgcgcgaggc
     4921 caagcgatct tcttcttgtc caagataagc ctgtctagct tcaagtatga cgggctgata
     4981 ctgggccggc aggcgctcca ttgcccagtc ggcagcgaca tccttcggcg cgattttgcc
     5041 ggttactgcg ctgtaccaaa tgcgggacaa cgtaagcact acatttcgct catcgccagc
     5101 ccagtcgggc ggcgagttcc atagcgttaa ggtttcattt agcgcctcaa atagatcctg
     5161 ttcaggaacc ggatcaaaga gttcctccgc cgctggacct accaaggcaa cgctatgttc
     5221 tcttgctttt gtcagcaaga tagccagatc aatgtcgatc gtggctggct cgaagatacc
     5281 tgcaagaatg tcattgcgct gccattctcc aaattgcagt tcgcgcttag ctggataacg
     5341 ccacggaatg atgtcgtcgt gcacaacaat ggtgacttct acagcgcgga gaatctcgct
     5401 ctctccaggg gaagccgaag tttccaaaag gtcgttgatc aaagctcgcc gcgttgtttc
     5461 atcaagcctt acggtcaccg taaccagcaa atcaatatca ctgtgtggct tcaggccgcc
     5521 atccactgcg gagccgtaca aatgtacggc cagcaacgtc ggttcgagat ggcgctcgat
     5581 gacgccaact acctctgata gttgagtcga tacttcggcg atcaccgctt ccctcatact
     5641 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     5701 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatcta agaaaccatt
     5761 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tcttcactta
     5821 tccctattac ttctaaatct gaggaggaaa atcgaatcgc taccttagga ccgttatagt
     5881 tactatcacc atatatgtaa ctggtgattc tttcgctacc ttagagtgca ccatagatca
     5941 acgacattac tatatatata atataggaag catttaatag aacagcatcg taatatatgt
     6001 gtactttgca gttatgacgc cagatggcag tagtggaaga tattctttat tgaaaaatag
     6061 cttgtcacct tacgtacaat cttgatccgg agcttttctt tttttgccga ttaagaatta
     6121 attcggtcga aaaaagaaaa ggagagggcc aagagggagg gcattggtga ctattgagca
     6181 cgtgagtata cgtgattaag cacacaaagg cagcttggag tatgtctgtt attaatttca
     6241 caggtagttc tggtccattg gtgaaagttt gcggcttgca gagcacagag gccgcagaat
     6301 gtgctctaga ttccgatgct gacttgctgg gtattatatg tgtgcccaat agaaagagaa
     6361 caattgaccc ggttattgca aggaaaattt caagtcttgt aaaagcatat aaaaatagtt
     6421 caggcactcc gaaatacttg gttggcgtgt ttcgtaatca acctaaggag gatgttttgg
     6481 ctctggtcaa tgattacggc attgatatcg tccaactgca tggagatgag tcgtggcaag
     6541 aataccaaga gttcctcggt ttgccagtta ttaaaagact cgtatttcca aaagactgca
     6601 acatactact cagtgcagct tcacagaaac ctcattcgtt tattcccttg tttgattcag
     6661 aagcaggtgg gacaggtgaa cttttggatt ggaactcgat ttctgactgg gttggaaggc
     6721 aagagagccc cgaaagctta cattttatgt tagctggtgg actgacgcca gaaaatgttg
     6781 gtgatgcgct tagattaaat ggcgttattg gtgttgatgt aagcggaggt gtggagacaa
     6841 atggtgtaaa agactctaac aaaatagcaa atttcgtcaa aaatgctaag aaataggtta
     6901 ttactgagta gtatttattt aagtattgtt tgtgcacttg ccgatctatg cggtgtgaaa
     6961 taccgcacag atgcgtaagg agaaaatacc gcatcaggaa attgtaagcg ttaatatttt
     7021 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     7081 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     7141 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     7201 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     7261 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     7321 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     7381 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     7441 gctacagggc gcgtccattc gccattcagg ctgcgcaact gttgggaagg gcgat
//
