LOCUS       40924_20940        7804 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Yeast expression vector pGAD424A20ZF-, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7804)
  AUTHORS   De Schamphelaire W, Olbrechts A, Meert J, Verhelst K, Roggeman 
            Fonseca M, Vanhoucke M, Beyaert R.
  TITLE     BCCM/LMBP Plasmid collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7804)
  AUTHORS   De Schamphelaire W.
  TITLE     Direct Submission
  JOURNAL   Submitted (02-FEB-2017) BCCM/LMBP, Universiteit Gent, 
            Technologiepark 927, 9052, BELGIUM
REFERENCE   3  (bases 1 to 7804)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7804)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (02-FEB-2017) BCCM/LMBP, Universiteit Gent, Technologiepark 927, 
            9052, BELGIUM"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7804
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        5..406
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             452..472
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
     CDS             488..829
                     /label=GAL4 activation domain
                     /note="activation domain of the GAL4 transcriptional
                     activator"
     CDS             845..871
                     /label=HA
                     /note="HA (human influenza hemagglutinin) epitope tag"
     CDS             887..1987
                     /codon_start=1
                     /note="unnamed protein product; hA20f"
                     /protein_id="SJL88290.1"
                     /translation="MAEQVLPQALYLSNMRKAVKIRERTPEDIFKPTNGIIHHFKTMHR
                     YTLEMFRTCQFCPQFREIIHKALIDRNIQATLESQKKLNWCREVRKLVALKTNGDGNCL
                     MHATSQYMWGVQDTDLVLRKALFSTLKETDTRNFKFRWQLESLKSQEFVETGLCYDTRN
                     WNDEWDNLIKMASTDTPMARSGLQYNSLEEIHIFVLCNILRRPIIVISDKMLRSLESGS
                     NFAPLKVGGIYLPLHWPAQECYRYPIVLGYDSHHFVPLVTLKDSGPEIRAVPLVNRDRG
                     RFEDLKVHFLTDPENEMKEKLLKEYLMVIEIPVQGWDHGTTHLINAAKLDEANLPKEIN
                     LVDDYFELVQHEYKKWQENSEQGRREG"
     terminator      2386..2573
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae
                     alcohol dehydrogenase 1 (ADH1) gene"
     CDS             complement(2694..3785)
                     /label=LEU2
                     /note="3-isopropylmalate dehydrogenase, required for
                     leucine biosynthesis"
     promoter        complement(3786..4190)
                     /label=LEU2 promoter
     primer_bind     complement(4232..4248)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4256..4272)
                     /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(4280..4310)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4325..4346)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4634..5222)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5396..6253)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(6254..6358)
                     /label=AmpR promoter
     rep_origin      6640..7804
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
ORIGIN
        1 gcttgcatgc aacttctttt cttttttttt cttttctctc tcccccgttg ttgtctcacc
       61 atatccgcaa tgacaaaaaa aatgatggaa gacactaaag gaaaaaatta acgacaaaga
      121 cagcaccaac agatgtcgtt gttccagagc tgatgagggg tatcttcgaa cacacgaaac
      181 tttttccttc cttcattcac gcacactact ctctaatgag caacggtata cggccttcct
      241 tccagttact tgaatttgaa ataaaaaaag tttgccgctt tgctatcaag tataaataga
      301 cctgcaatta ttaatctttt gtttcctcgt cattgttctc gttccctttc ttccttgttt
      361 ctttttctgc acaatatttc aagctatacc aagcatacaa tcaactccaa gctttgcaaa
      421 gatggataaa gcggaattaa ttcccgagcc tccaaaaaag aagagaaagg tcgaattggg
      481 taccgccgcc aattttaatc aaagtgggaa tattgctgat agctcattgt ccttcacttt
      541 cactaacagt agcaacggtc cgaacctcat aacaactcaa acaaattctc aagcgctttc
      601 acaaccaatt gcctcctcta acgttcatga taacttcatg aataatgaaa tcacggctag
      661 taaaattgat gatggtaata attcaaaacc actgtcacct ggttggacgg accaaactgc
      721 gtataacgcg tttggaatca ctacagggat gtttaatacc actacaatgg atgatgtata
      781 taactatcta ttcgatgatg aagatacccc accaaaccca aaaaaagaga tcgaattcat
      841 ggcttaccca tacgatgttc cagattacgc tagcttgggt ggtcatatgg ctgaacaagt
      901 ccttcctcag gctttgtatt tgagcaatat gcggaaagct gtgaagatac gggagagaac
      961 tccagaagac atttttaaac ctactaatgg gatcattcat cattttaaaa ccatgcaccg
     1021 atacacactg gaaatgttca gaacttgcca gttttgtcct cagtttcggg agatcatcca
     1081 caaagccctc atcgacagaa acatccaggc caccctggaa agccagaaga aactcaactg
     1141 gtgtcgagaa gtccggaagc ttgtggcgct gaaaacgaac ggtgacggca attgcctcat
     1201 gcatgccact tctcagtaca tgtggggcgt tcaggacaca gacttggtac tgaggaaggc
     1261 gctgttcagc acgctcaagg aaacagacac acgcaacttt aaattccgct ggcaactgga
     1321 gtctctcaaa tctcaggaat ttgttgaaac ggggctttgc tatgatactc ggaactggaa
     1381 tgatgaatgg gacaatctta tcaaaatggc ttccacagac acacccatgg cccgaagtgg
     1441 acttcagtac aactcactgg aagaaataca catatttgtc ctttgcaaca tcctcagaag
     1501 gccaatcatt gtcatttcag acaaaatgct aagaagtttg gaatcaggtt ccaatttcgc
     1561 ccctttgaaa gtgggtggaa tttacttgcc tctccactgg cctgcccagg aatgctacag
     1621 ataccccatt gttctcggct atgacagcca tcattttgta cccttggtga ccctgaagga
     1681 cagtgggcct gaaatccgag ctgttccact tgttaacaga gaccggggaa gatttgaaga
     1741 cttaaaagtt cactttttga cagatcctga aaatgagatg aaggagaagc tcttaaaaga
     1801 gtacttaatg gtgatagaaa tccccgtcca aggctgggac catggcacaa ctcatctcat
     1861 caatgccgca aagttggatg aagctaactt accaaaagaa atcaatctgg tagatgatta
     1921 ctttgaactt gttcagcatg agtacaagaa atggcaggaa aacagcgagc aggggaggag
     1981 agagggggga tccgtcgacc tgcagagatc tatgaatcgt agatactgaa aaaccccgca
     2041 agttcacttc aactgtgcat cgtgcaccat ctcaatttct ttcatttata catcgttttg
     2101 ccttctttta tgtaactata ctcctctaag tttcaatctt ggccatgtaa cctctgatct
     2161 atagaatttt ttaaatgact agaattaatg cccatctttt ttttggacct aaattcttca
     2221 tgaaaatata ttacgagggc ttattcagaa gctttggact tcttcgccag aggtttggtc
     2281 aagtctccaa tcaaggttgt cggcttgtct accttgccag aaatttacga aaagatggaa
     2341 aagggtcaaa tcgttggtag atacgttgtt gacacttcta aataagcgaa tttcttatga
     2401 tttatgattt ttattattaa ataagttata aaaaaaataa gtgtatacaa attttaaagt
     2461 gactcttagg ttttaaaacg aaaattcttg ttcttgagta actctttcct gtaggtcagg
     2521 ttgctttctc aggtatagca tgaggtcgct cttattgacc acacctctac cggcatgccc
     2581 gaaattcccc taccctatga acatattcca ttttgtaatt tcgtgtcgtt tctattatga
     2641 atttcattta taaagtttat gtacaaatat cataaaaaaa gagaatcttt ttaagcaagg
     2701 attttcttaa cttcttcggc gacagcatca ccgacttcgg tggtactgtt ggaaccacct
     2761 aaatcaccag ttctgatacc tgcatccaaa acctttttaa ctgcatcttc aatggcctta
     2821 ccttcttcag gcaagttcaa tgacaatttc aacatcattg cagcagacaa gatagtggcg
     2881 atagggtcaa ccttattctt tggcaaatct ggagcagaac cgtggcatgg ttcgtacaaa
     2941 ccaaatgcgg tgttcttgtc tggcaaagag gccaaggacg cagatggcaa caaacccaag
     3001 gaacctggga taacggaggc ttcatcggag atgatatcac caaacatgtt gctggtgatt
     3061 ataataccat ttaggtgggt tgggttctta actaggatca tggcggcaga atcaatcaat
     3121 tgatgttgaa ccttcaatgt aggaaattcg ttcttgatgg tttcctccac agtttttctc
     3181 cataatcttg aagaggccaa aacattagct ttatccaagg accaaatagg caatggtggc
     3241 tcatgttgta gggccatgaa agcggccatt cttgtgattc tttgcacttc tggaacggtg
     3301 tattgttcac tatcccaagc gacaccatca ccatcgtctt cctttctctt accaaagtaa
     3361 atacctccca ctaattctct gacaacaacg aagtcagtac ctttagcaaa ttgtggcttg
     3421 attggagata agtctaaaag agagtcggat gcaaagttac atggtcttaa gttggcgtac
     3481 aattgaagtt ctttacggat ttttagtaaa ccttgttcag gtctaacact acctgtaccc
     3541 catttaggac cacccacagc acctaacaaa acggcatcaa ccttcttgga ggcttccagc
     3601 gcctcatctg gaagtgggac acctgtagca tcgatagcag caccaccaat taaatgattt
     3661 tcgaaatcga acttgacatt ggaacgaaca tcagaaatag ctttaagaac cttaatggct
     3721 tcggctgtga tttcttgacc aacgtggtca cctggcaaaa cgacgatctt cttaggggca
     3781 gacattagaa tggtatatcc ttgaaatata tatatatatt gctgaaatgt aaaaggtaag
     3841 aaaagttaga aagtaagacg attgctaacc acctattgga aaaaacaata ggtccttaaa
     3901 taatattgtc aacttcaagt attgtgatgc aagcatttag tcatgaacgc ttctctattc
     3961 tatatgaaaa gccggttccg gcctctcacc tttccttttt ctcccaattt ttcagttgaa
     4021 aaaggtatat gcgtcaggcg acctctgaaa ttaacaaaaa atttccagtc atcgaatttg
     4081 attctgtgcg atagcgcccc tgtgtgttct cgttatgttg aggaaaaaaa taatggttgc
     4141 taagagattc gaactcttgc atcttacgat acctgagtat tcccacagtt ggggatctcg
     4201 actctagcta gaggatcaat tcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     4261 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     4321 cctaatgagt gaggtaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     4381 gaaacctgtc gtgccagctg gattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     4441 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     4501 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     4561 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     4621 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     4681 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     4741 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     4801 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     4861 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     4921 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     4981 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     5041 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     5101 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     5161 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     5221 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     5281 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     5341 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     5401 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     5461 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     5521 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     5581 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     5641 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     5701 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     5761 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     5821 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     5881 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     5941 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     6001 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     6061 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     6121 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     6181 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     6241 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     6301 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     6361 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
     6421 ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa
     6481 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
     6541 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact
     6601 atgcggcatc agagcagatt gtactgagag tgcaccataa cgcatttaag cataaacacg
     6661 cactatgccg ttcttctcat gtatatatat atacaggcaa cacgcagata taggtgcgac
     6721 gtgaacagtg agctgtatgt gcgcagctcg cgttgcattt tcggaagcgc tcgttttcgg
     6781 aaacgctttg aagttcctat tccgaagttc ctattctcta gctagaaagt ataggaactt
     6841 cagagcgctt ttgaaaacca aaagcgctct gaagacgcac tttcaaaaaa ccaaaaacgc
     6901 accggactgt aacgagctac taaaatattg cgaataccgc ttccacaaac attgctcaaa
     6961 agtatctctt tgctatatat ctctgtgcta tatccctata taacctaccc atccaccttt
     7021 cgctccttga acttgcatct aaactcgacc tctacatttt ttatgtttat ctctagtatt
     7081 actctttaga caaaaaaatt gtagtaagaa ctattcatag agtgaatcga aaacaatacg
     7141 aaaatgtaaa catttcctat acgtagtata tagagacaaa atagaagaaa ccgttcataa
     7201 ttttctgacc aatgaagaat catcaacgct atcactttct gttcacaaag tatgcgcaat
     7261 ccacatcggt atagaatata atcggggatg cctttatctt gaaaaaatgc acccgcagct
     7321 tcgctagtaa tcagtaaacg cgggaagtgg agtcaggctt tttttatgga agagaaaata
     7381 gacaccaaag tagccttctt ctaaccttaa cggacctaca gtgcaaaaag ttatcaagag
     7441 actgcattat agagcgcaca aaggagaaaa aaagtaatct aagatgcttt gttagaaaaa
     7501 tagcgctctc gggatgcatt tttgtagaac aaaaaagaag tatagattct ttgttggtaa
     7561 aatagcgctc tcgcgttgca tttctgttct gtaaaaatgc agctcagatt ctttgtttga
     7621 aaaattagcg ctctcgcgtt gcatttttgt tttacaaaaa tgaagcacag attcttcgtt
     7681 ggtaaaatag cgctttcgcg ttgcatttct gttctgtaaa aatgcagctc agattctttg
     7741 tttgaaaaat tagcgctctc gcgttgcatt tttgttctac aaaatgaagc acagatgctt
     7801 cgtt
//
