LOCUS       40924_20935        7953 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 7953)
  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  JOURNAL   Unpublished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 7953)
  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 7953)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7953)
  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            "Unpublished"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..7953
                     /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             848..2104
                     /codon_start=1
                     /note="unnamed protein product; hA20f"
                     /protein_id="SJL88294.1"
                     /translation="MEPSVPQLSLMDVKCETPNCPFFMSVNTQPLCHECSERRQKNQNK
                     LPKLNSKPGPEGLPGMALGASRGEAYEPLAWNPEESTGGPHSAPPTAPSPFLFSETTAM
                     KCRSPGCPFTLNVQHNGFCERCHNARQLHASHAPDHTRHLDPGKCQACLQDVTRTFNGI
                     CSTCFKRTTAEASSSLSTSLPPSCHQRSKSDPSRLVRSPSPHSCHRAGNDAPAGCLSQA
                     ARTPGDRTGTSKCRKAGCVYFGTPENKGFCTLCFIEYRENKHFAAASGKVSPTASRFQN
                     TIPCLGRECGTLGSTMFEGYCQKCFIEAQNQRFHEAKRTEEQLRSSQRRDVPRTTQSTS
                     RPKCARASCKNILACRSEELCMECQHPNQRMGPGAHRGEPAPEDPPKQRCRAPACDHFG
                     NAKCNGYCNECFQFKQMYG"
     terminator      2535..2722
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae
                     alcohol dehydrogenase 1 (ADH1) gene"
     CDS             complement(2843..3934)
                     /label=LEU2
                     /note="3-isopropylmalate dehydrogenase, required for
                     leucine biosynthesis"
     promoter        complement(3935..4339)
                     /label=LEU2 promoter
     primer_bind     complement(4381..4397)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4405..4421)
                     /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(4429..4459)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4474..4495)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4783..5371)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5545..6402)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(6403..6507)
                     /label=AmpR promoter
     rep_origin      6789..7953
                     /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 tcgaattcgg
      841 tgcggccatg gaaccttccg tgccccagct ttctctcatg gatgtaaaat gtgaaacgcc
      901 caactgcccc ttcttcatgt ctgtgaacac ccagccttta tgccatgagt gctcagagag
      961 gcggcaaaag aatcaaaaca aactcccaaa gctgaactcc aagccgggcc ctgaggggct
     1021 ccctggcatg gcgctcgggg cctctcgggg agaagcctat gagcccttgg cgtggaaccc
     1081 tgaggagtcc actggggggc ctcattcggc cccaccgaca gcacccagcc cttttctgtt
     1141 cagtgagacc actgccatga agtgcaggag ccccggctgc cccttcacac tgaatgtgca
     1201 gcacaacgga ttttgtgaac gttgccacaa cgcccggcaa cttcacgcca gccacgcccc
     1261 agaccacaca aggcacttgg atcccgggaa gtgccaagcc tgcctccagg atgttaccag
     1321 gacatttaat gggatctgca gtacttgctt caaaaggact acagcagagg cctcctccag
     1381 cctcagcacc agcctccctc cttcctgtca ccagcgttcc aagtcagatc cctcgcggct
     1441 cgtccggagc ccctccccgc attcttgcca cagagctgga aacgacgccc ctgctggctg
     1501 cctgtctcaa gctgcacgga ctcctgggga caggacgggg acgagcaagt gcagaaaagc
     1561 cggctgcgtg tattttggga ctccagaaaa caagggcttt tgcacactgt gtttcatcga
     1621 gtacagagaa aacaaacatt ttgctgctgc ctcagggaaa gtcagtccca cagcgtccag
     1681 gttccagaac accattccgt gcctggggag ggaatgcggc acccttggaa gcaccatgtt
     1741 tgaaggatac tgccagaagt gtttcattga agctcagaat cagagatttc atgaggccaa
     1801 aaggacagaa gagcaactga gatcgagcca gcgcagagat gtgcctcgaa ccacacaaag
     1861 cacctcaagg cccaagtgcg cccgggcctc ctgcaagaac atcctggcct gccgcagcga
     1921 ggagctctgc atggagtgtc agcatcccaa ccagaggatg ggccctgggg cccaccgggg
     1981 tgagcctgcc cccgaagacc cccccaagca gcgttgccgg gcccccgcct gtgatcattt
     2041 tggcaatgcc aagtgcaacg gctactgcaa cgaatgcttt cagttcaagc agatgtatgg
     2101 ctaaccggaa acaggtgggt cacctcctgc aagaagtggg gcctcgacct gcagagatct
     2161 atgaatcgta gatactgaaa aaccccgcaa gttcacttca actgtgcatc gtgcaccatc
     2221 tcaatttctt tcatttatac atcgttttgc cttcttttat gtaactatac tcctctaagt
     2281 ttcaatcttg gccatgtaac ctctgatcta tagaattttt taaatgacta gaattaatgc
     2341 ccatcttttt tttggaccta aattcttcat gaaaatatat tacgagggct tattcagaag
     2401 ctttggactt cttcgccaga ggtttggtca agtctccaat caaggttgtc ggcttgtcta
     2461 ccttgccaga aatttacgaa aagatggaaa agggtcaaat cgttggtaga tacgttgttg
     2521 acacttctaa ataagcgaat ttcttatgat ttatgatttt tattattaaa taagttataa
     2581 aaaaaataag tgtatacaaa ttttaaagtg actcttaggt tttaaaacga aaattcttgt
     2641 tcttgagtaa ctctttcctg taggtcaggt tgctttctca ggtatagcat gaggtcgctc
     2701 ttattgacca cacctctacc ggcatgcccg aaattcccct accctatgaa catattccat
     2761 tttgtaattt cgtgtcgttt ctattatgaa tttcatttat aaagtttatg tacaaatatc
     2821 ataaaaaaag agaatctttt taagcaagga ttttcttaac ttcttcggcg acagcatcac
     2881 cgacttcggt ggtactgttg gaaccaccta aatcaccagt tctgatacct gcatccaaaa
     2941 cctttttaac tgcatcttca atggccttac cttcttcagg caagttcaat gacaatttca
     3001 acatcattgc agcagacaag atagtggcga tagggtcaac cttattcttt ggcaaatctg
     3061 gagcagaacc gtggcatggt tcgtacaaac caaatgcggt gttcttgtct ggcaaagagg
     3121 ccaaggacgc agatggcaac aaacccaagg aacctgggat aacggaggct tcatcggaga
     3181 tgatatcacc aaacatgttg ctggtgatta taataccatt taggtgggtt gggttcttaa
     3241 ctaggatcat ggcggcagaa tcaatcaatt gatgttgaac cttcaatgta ggaaattcgt
     3301 tcttgatggt ttcctccaca gtttttctcc ataatcttga agaggccaaa acattagctt
     3361 tatccaagga ccaaataggc aatggtggct catgttgtag ggccatgaaa gcggccattc
     3421 ttgtgattct ttgcacttct ggaacggtgt attgttcact atcccaagcg acaccatcac
     3481 catcgtcttc ctttctctta ccaaagtaaa tacctcccac taattctctg acaacaacga
     3541 agtcagtacc tttagcaaat tgtggcttga ttggagataa gtctaaaaga gagtcggatg
     3601 caaagttaca tggtcttaag ttggcgtaca attgaagttc tttacggatt tttagtaaac
     3661 cttgttcagg tctaacacta cctgtacccc atttaggacc acccacagca cctaacaaaa
     3721 cggcatcaac cttcttggag gcttccagcg cctcatctgg aagtgggaca cctgtagcat
     3781 cgatagcagc accaccaatt aaatgatttt cgaaatcgaa cttgacattg gaacgaacat
     3841 cagaaatagc tttaagaacc ttaatggctt cggctgtgat ttcttgacca acgtggtcac
     3901 ctggcaaaac gacgatcttc ttaggggcag acattagaat ggtatatcct tgaaatatat
     3961 atatatattg ctgaaatgta aaaggtaaga aaagttagaa agtaagacga ttgctaacca
     4021 cctattggaa aaaacaatag gtccttaaat aatattgtca acttcaagta ttgtgatgca
     4081 agcatttagt catgaacgct tctctattct atatgaaaag ccggttccgg cctctcacct
     4141 ttcctttttc tcccaatttt tcagttgaaa aaggtatatg cgtcaggcga cctctgaaat
     4201 taacaaaaaa tttccagtca tcgaatttga ttctgtgcga tagcgcccct gtgtgttctc
     4261 gttatgttga ggaaaaaaat aatggttgct aagagattcg aactcttgca tcttacgata
     4321 cctgagtatt cccacagttg gggatctcga ctctagctag aggatcaatt cgtaatcatg
     4381 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     4441 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg aggtaactca cattaattgc
     4501 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgg attaatgaat
     4561 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     4621 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     4681 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     4741 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     4801 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     4861 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     4921 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     4981 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     5041 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     5101 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     5161 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     5221 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     5281 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     5341 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     5401 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     5461 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     5521 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     5581 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     5641 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     5701 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     5761 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     5821 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     5881 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     5941 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     6001 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     6061 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     6121 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     6181 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     6241 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     6301 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     6361 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     6421 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     6481 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     6541 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     6601 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     6661 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     6721 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
     6781 gcaccataac gcatttaagc ataaacacgc actatgccgt tcttctcatg tatatatata
     6841 tacaggcaac acgcagatat aggtgcgacg tgaacagtga gctgtatgtg cgcagctcgc
     6901 gttgcatttt cggaagcgct cgttttcgga aacgctttga agttcctatt ccgaagttcc
     6961 tattctctag ctagaaagta taggaacttc agagcgcttt tgaaaaccaa aagcgctctg
     7021 aagacgcact ttcaaaaaac caaaaacgca ccggactgta acgagctact aaaatattgc
     7081 gaataccgct tccacaaaca ttgctcaaaa gtatctcttt gctatatatc tctgtgctat
     7141 atccctatat aacctaccca tccacctttc gctccttgaa cttgcatcta aactcgacct
     7201 ctacattttt tatgtttatc tctagtatta ctctttagac aaaaaaattg tagtaagaac
     7261 tattcataga gtgaatcgaa aacaatacga aaatgtaaac atttcctata cgtagtatat
     7321 agagacaaaa tagaagaaac cgttcataat tttctgacca atgaagaatc atcaacgcta
     7381 tcactttctg ttcacaaagt atgcgcaatc cacatcggta tagaatataa tcggggatgc
     7441 ctttatcttg aaaaaatgca cccgcagctt cgctagtaat cagtaaacgc gggaagtgga
     7501 gtcaggcttt ttttatggaa gagaaaatag acaccaaagt agccttcttc taaccttaac
     7561 ggacctacag tgcaaaaagt tatcaagaga ctgcattata gagcgcacaa aggagaaaaa
     7621 aagtaatcta agatgctttg ttagaaaaat agcgctctcg ggatgcattt ttgtagaaca
     7681 aaaaagaagt atagattctt tgttggtaaa atagcgctct cgcgttgcat ttctgttctg
     7741 taaaaatgca gctcagattc tttgtttgaa aaattagcgc tctcgcgttg catttttgtt
     7801 ttacaaaaat gaagcacaga ttcttcgttg gtaaaatagc gctttcgcgt tgcatttctg
     7861 ttctgtaaaa atgcagctca gattctttgt ttgaaaaatt agcgctctcg cgttgcattt
     7921 ttgttctaca aaatgaagca cagatgcttc gtt
//
