LOCUS       Exported                9637 bp ds-DNA     circular SYN 06-AUG-2018
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pGADCg
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9637)
  AUTHORS   Stellberger T, Hauser R, Baiker A, Pothineni VR, Haas J, Uetz P
  TITLE     Improving the yeast two-hybrid system with permutated fusions 
            proteins: the Varicella Zoster Virus interactome
  JOURNAL   Proteome Sci. 2010 Feb 15;8:8.
REFERENCE   2  (bases 1 to 9637)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 6, 2018 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9143
                     /dnas_title="pCSJ182"
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        771..1476
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             1522..1542
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian
                     virus 40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    1561..1587
                     /note="attB1"
     CDS             2785..3126
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="activation domain of the GAL4 transcriptional
                     activator"
                     /label=GAL4 activation domain
                     /translation="ANFNQSGNIADSSLSFTFTNSSNGPNLITTQTNSQALSQPIASS
                     NVHDNFMNNEITASKIDDGNNSKPLSPGWTDQTAYNAFGITTGMFNTTTMDDVYNYLF
                     DDEDTPPNPKKE"
     terminator      3571..3758
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(3626..3650)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(3875..4969)
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENH
                     LIGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQL
                     YANLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSE
                     QYTVPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLK
                     VQHQLIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPD
                     KNTAFGLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLD
                     AGIRTGDLGGSNSTTEVGDAVAEEVKKILA"
     promoter        complement(4970..5375)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     primer_bind     complement(5417..5433)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar
                     variants"
     primer_bind     complement(5417..5433)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5430..5452)
                     /label="M13/pUC Reverse"
                     /note="In lacZ gene"
     protein_bind    5441..5457
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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(5465..5495)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5510..5531
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the
                     presence of cAMP."
     protein_bind    complement(5586..5619)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core
                     sequence (GCATACAT)."
     protein_bind    complement(5697..5730)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core
                     sequence (GCATACAT)."
     primer_bind     complement(5799..5816)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5970..6558)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     primer_bind     complement(6050..6069)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6729..7589)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin,
                     and related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW"
     primer_bind     7352..7371
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7590..7694)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7762..7780
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward
                     primer"
     primer_bind     complement(7818..7840)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7940..7959
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable
                     marker"
     rep_origin      7976..9140
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    2746..2771
                     /note="attB2"
     CDS             2680..2745
                     /note="3xflag"
     CDS             1588..2673
                     /note="GID4"
ORIGIN      
        1 tgcatgcctg caggtcgaga tccgggatcg aagaaatgat ggtaaatgaa ataggaaatc
       61 aaggagcatg aaggcaaaag acaaatataa gggtcgaacg aaaaataaag tgaaaagtgt
      121 tgatatgatg tatttggctt tgcggcgccg aaaaaacgag tttacgcaat tgcacaatca
      181 tgctgactct gtggcggacc cgcgctcttg ccggcccggc gataacgctg ggcgtgaggc
      241 tgtgcccggc ggagtttttt gcgcctgcat tttccaaggt ttaccctgcg ctaaggggcg
      301 agattggaga agcaataaga atgccggttg gggttgcgat gatgacgacc acgacaactg
      361 gtgtcattat ttaagttgcc gaaagaacct gagtgcattt gcaacatgag tatactagaa
      421 gaatgagcca agacttgcga gacgcgagtt tgccggtggt gcgaacaata gagcgaccat
      481 gaccttgaag gtgagacgcg cataaccgct agagtacttt gaagaggaaa cagcaatagg
      541 gttgctacca gtataaatag acaggtacat acaacactgg aaatggttgt ctgtttgagt
      601 acgctttcaa ttcatttggg tgtgcacttt attatgttac aatatggaag ggaactttac
      661 acttctccta tgcacatata ttaattaaag tccaatgcta gtagagaagg ggggtaacac
      721 ccctccgcgc tcttttccga tttttttcta aaccgtggaa tatttcggat atccttttgt
      781 tgtttccggg tgtacaatat ggacttcctc ttttctggca accaaaccca tacatcggga
      841 ttcctataat accttcgttg gtctccctaa catgtaggtg gcggagggga gatatacaat
      901 agaacagata ccagacaaga cataatgggc taaacaagac tacaccaatt acactgcctc
      961 attgatggtg gtacataacg aactaatact gtagccctag acttgatagc catcatcata
     1021 tcgaagtttc actacccttt ttccatttgc catctattga agtaataata ggcgcatgca
     1081 acttcttttc tttttttttc ttttctctct cccccgttgt tgtctcacca tatccgcaat
     1141 gacaaaaaaa atgatggaag acactaaagg aaaaaattaa cgacaaagac agcaccaaca
     1201 gatgtcgttg ttccagagct gatgaggggt atctcgaagc acacgaaact ttttccttcc
     1261 ttcattcacg cacactactc tctaatgagc aacggtatac ggccttcctt ccagttactt
     1321 gaatttgaaa taaaaaaaag tttgctgtct tgctatcaag tataaataga cctgcaatta
     1381 ttaatctttt gtttcctcgt cattgttctc gttccctttc ttccttgttt ctttttctgc
     1441 acaatatttc aagctatacc aagcatacaa tcaactccaa gctttgcaaa gatggataaa
     1501 gcggaattaa ttcccgagcc tccaaaaaag aagagaaagg tcgaattggg taccgcatca
     1561 acaagtttgt acaaaaaagc aggcttcatg atcaataatc ctaaggtaga cagtgtagcg
     1621 gagaaaccca aagctgtgac atcaaagcag tcggagcaag cggcttcgcc agaaccaaca
     1681 ccagcccccc ctgtttctag aaatcagtat ccgatcacgt tcaacttgac ttcaaccgca
     1741 ccctttcatc ttcatgaccg tcatcgctac ttacaagagc aagatcttta caagtgcgct
     1801 tctagggatt cgttgtcttc cctgcagcaa ctcgcccata cacctaatgg gtcgacaagg
     1861 aagaaatata ttgttgagga ccaatctccc tatagttcgg aaaatccagt cattgtgacc
     1921 tcttcctata accatacggt ttgcacaaac tacttaagac caagaatgca gtttacaggg
     1981 taccagatat caggatacaa acgctatcag gtaacagtta acttaaagac tgtagatttg
     2041 ccaaagaaag attgcacgtc gctgtcccct catttatctg gatttttgtc gataagagga
     2101 ctcacgaacc aacacccgga aatcagtaca tattttgaag cctacgcggt aaaccacaag
     2161 gaattagggt tcttgtcctc tagctggaaa gatgaacctg ttttaaacga attcaaagcc
     2221 acagaccaaa cagacttaga acactggata aatttcccct cctttagaca gcttttcctg
     2281 atgagccaaa aaaacggtct caactcaact gacgacaatg gcactacaaa tgcagccaag
     2341 aagttgcctc cacagcagct tcctactaca ccaagcgcag acgctggtaa catatcaaga
     2401 atttttagcc aagagaaaca atttgacaac tacttgaacg aacggtttat atttatgaaa
     2461 tggaaggaaa aatttttggt accagatgcc ctattaatgg aaggtgtaga cggcgcatct
     2521 tatgatgggt tttattacat tgtccatgat caagttaccg ggaacattca agggttttac
     2581 tatcatcaag atgctgaaaa gttccaacag ctggaattag taccatcttt gaaaaataaa
     2641 gtcgagtcca gtgattgttc ttttgagttt gctCCCGGGG ATTACAAGGA TCATGATGGT
     2701 GATTACAAGG ATCATGACAT TGACTATAAG GATGATGATG ACAAAGACCC AGCTTTCTTG
     2761 TACAAAGTGG Ttgatggtac cgccgccaat tttaatcaaa gtgggaatat tgctgatagc
     2821 tcattgtcct tcactttcac taacagtagc aacggtccga acctcataac aactcaaaca
     2881 aattctcaag cgctttcaca accaattgcc tcctctaacg ttcatgataa cttcatgaat
     2941 aatgaaatca cggctagtaa aattgatgat ggtaataatt caaaaccact gtcacctggt
     3001 tggacggacc aaactgcgta taacgcgttt ggaatcacta cagggatgtt taataccact
     3061 acaatggatg atgtatataa ctatctattc gatgatgaag ataccccacc aaacccaaaa
     3121 aaagagatcc tcgagtacga cgtaccagat tacgcttagt gatcacgata cgggatccat
     3181 cgagctcgag ctgcagatga atcgtagata ctgaaaaacc ccgcaagttc acttcaactg
     3241 tgcatcgtgc accatctcaa tttctttcat ttatacatcg ttttgccttc ttttatgtaa
     3301 ctatactcct ctaagtttca atcttggcca tgtaacctct gatctataga attttttaaa
     3361 tgactagaat taatgcccat cttttttttg gacctaaatt cttcatgaaa atatattacg
     3421 agggcttatt cagaagcttt ggacttcttc gccagaggtt tggtcaagtc tccaatcaag
     3481 gttgtcggct tgtctacctt gccagaaatt tacgaaaaga tggaaaaggg tcaaatcgtt
     3541 ggtagatacg ttgttgacac ttctaaataa gcgaatttct tatgatttat gatttttatt
     3601 attaaataag ttataaaaaa aataagtgta tacaaatttt aaagtgactc ttaggtttta
     3661 aaacgaaaat tcttattctt gagtaactct ttcctgtagg tcaggttgct ttctcaggta
     3721 tagcatgagg tcgctcttat tgaccacacc tctaccggcc ggtcgaaatt cccctaccct
     3781 atgaacatat tccattttgt aatttcgtgt cgtttctatt atgaatttca tttataaagt
     3841 ttatgtacaa atatcataaa aaaagagaat ctttttaagc aaggattttc ttaacttctt
     3901 cggcgacagc atcaccgact tcggtggtac tgttggaacc acctaaatca ccagttctga
     3961 tacctgcatc caaaaccttt ttaactgcat cttcaatggc cttaccttct tcaggcaagt
     4021 tcaatgacaa tttcaacatc attgcagcag acaagatagt ggcgataggg ttgaccttat
     4081 tctttggcaa atctggagca gaaccgtggc atggttcgta caaaccaaat gcggtgttct
     4141 tgtctggcaa agaggccaag gacgcagatg gcaacaaacc caaggaacct gggataacgg
     4201 aggcttcatc ggagatgata tcaccaaaca tgttgctggt gattataata ccatttaggt
     4261 gggttgggtt cttaactagg atcatggcgg cagaatcaat caattgatgt tgaaccttca
     4321 atgtaggaaa ttcgttcttg atggtttcct ccacagtttt tctccataat cttgaagagg
     4381 ccaaaacatt agctttatcc aaggaccaaa taggcaatgg tggctcatgt tgtagggcca
     4441 tgaaagcggc cattcttgtg attctttgca cttctggaac ggtgtattgt tcactatccc
     4501 aagcgacacc atcaccatcg tcttcctttc tcttaccaaa gtaaatacct cccactaatt
     4561 ctctgacaac aacgaagtca gtacctttag caaattgtgg cttgattgga gataagtcta
     4621 aaagagagtc ggatgcaaag ttacatggtc ttaagttggc gtacaattga agttctttac
     4681 ggatttttag taaaccttgt tcaggtctaa cactacctgt accccattta ggaccaccca
     4741 cagcacctaa caaaacggca tcaaccttct tggaggcttc cagcgcctca tctggaagtg
     4801 ggacacctgt agcgtcgata gcagcaccac caattaaatg attttcgaaa tcgaacttga
     4861 cattggaacg aacatcagaa atagctttaa gaaccttaat ggcttcggct gtgatttctt
     4921 gaccaacgtg gtcacctggc aaaacgacga tcttcttagg ggcagacatt agaatggtat
     4981 atccttgaaa tatatatata tattgctgaa atgtaaaagg taagaaaagt tagaaagtaa
     5041 gacgattgct aaccacctat tggaaaaaac aataggtcct taaataatat tgtcaacttc
     5101 aagtattgtg atgcaagcat ttagtcatga acgcttctct attctatatg aaaagccggt
     5161 tccggcgctc tcacctttcc tttttctccc aatttttcag ttgaaaaagg tatatgcgtc
     5221 aggcgacctc tgaaattaac aaaaaatttc cagtcatcga atttgattct gtgcgatagc
     5281 gcccctgtgt gttctcgtta tgttgaggaa aaaaataatg gttgctaaga gattcgaact
     5341 cttgcatctt acgatacctg agtattccca cagttgggga tctcgactct agctagagga
     5401 tcaattcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     5461 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     5521 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     5581 agctgataac ttcgtataat gtatgctata cgaagttatt aggtctgaag aggagtttac
     5641 gtccagccaa gctagcttgg ctgcaggtcg agcggccgcg atccggaacc cttaatataa
     5701 cttcgtataa tgtatgctat acgaagttat cagctgcatt aatgaatcgg ccaacgcgcg
     5761 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     5821 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     5881 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     5941 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     6001 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     6061 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     6121 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     6181 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     6241 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     6301 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     6361 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     6421 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     6481 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     6541 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     6601 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     6661 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     6721 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     6781 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     6841 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     6901 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     6961 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     7021 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     7081 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     7141 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     7201 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     7261 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     7321 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     7381 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     7441 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     7501 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     7561 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     7621 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     7681 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt
     7741 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc
     7801 ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg
     7861 taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt
     7921 cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccataacgca
     7981 tttaagcata aacacgcact atgccgttct tctcatgtat atatatatac aggcaacacg
     8041 cagatatagg tgcgacgtga acagtgagct gtatgtgcgc agctcgcgtt gcattttcgg
     8101 aagcgctcgt tttcggaaac gctttgaagt tcctattccg aagttcctat tctctagcta
     8161 gaaagtatag gaacttcaga gcgcttttga aaaccaaaag cgctctgaag acgcactttc
     8221 aaaaaaccaa aaacgcaccg gactgtaacg agctactaaa atattgcgaa taccgcttcc
     8281 acaaacattg ctcaaaagta tctctttgct atatatctct gtgctatatc cctatataac
     8341 ctacccatcc acctttcgct ccttgaactt gcatctaaac tcgacctcta cattttttat
     8401 gtttatctct agtattactc tttagacaaa aaaattgtag taagaactat tcatagagtg
     8461 aatcgaaaac aatacgaaaa tgtaaacatt tcctatacgt agtatataga gacaaaatag
     8521 aagaaaccgt tcataatttt ctgaccaatg aagaatcatc aacgctatca ctttctgttc
     8581 acaaagtatg cgcaatccac atcggtatag aatataatcg gggatgcctt tatcttgaaa
     8641 aaatgcaccc gcagcttcgc tagtaatcag taaacgcggg aagtggagtc aggctttttt
     8701 tatggaagag aaaatagaca ccaaagtagc cttcttctaa ccttaacgga cctacagtgc
     8761 aaaaagttat caagagactg cattatagag cgcacaaagg agaaaaaaag taatctaaga
     8821 tgctttgtta gaaaaatagc gctctcggga tgcatttttg tagaacaaaa aagaagtata
     8881 gattctttgt tggtaaaata gcgctctcgc gttgcatttc tgttctgtaa aaatgcagct
     8941 cagattcttt gtttgaaaaa ttagcgctct cgcgttgcat ttttgtttta caaaaatgaa
     9001 gcacagattc ttcgttggta aaatagcgct ttcgcgttgc atttctgttc tgtaaaaatg
     9061 cagctcagat tctttgtttg aaaaattagc gctctcgcgt tgcatttttg ttctacaaaa
     9121 tgaagcacag atgcttcgtt gct
//

