LOCUS       40924_14390        9677 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Yeast vector pDEST-GADT7, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9677)
  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  JOURNAL   Unpublished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 9677)
  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 9677)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 9677)
  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            "Unpublished"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..9677
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        771..1476
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             1522..1542
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
     CDS             1558..1899
                     /label=GAL4 activation domain
                     /note="activation domain of the GAL4 transcriptional
                     activator"
     promoter        1905..1923
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1930..1939
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1942..1968
                     /label=HA
                     /note="HA (human influenza hemagglutinin) epitope tag"
     protein_bind    1999..2123
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        2148..2178
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             2232..2888
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     CDS             3233..3535
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     protein_bind    complement(3579..3703)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     terminator      4105..4292
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae
                     alcohol dehydrogenase 1 (ADH1) gene"
     CDS             complement(4412..5503)
                     /label=LEU2
                     /note="3-isopropylmalate dehydrogenase, required for
                     leucine biosynthesis"
     promoter        complement(5504..5909)
                     /label=LEU2 promoter
     primer_bind     complement(5951..5967)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5975..5991)
                     /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(5999..6029)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(6044..6065)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     protein_bind    6120..6153
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    complement(6231..6264)
                     /label=loxP
                     /note="loxP"
     protein_bind    complement(6231..6264)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     rep_origin      complement(6504..7092)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7266..8123)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(8124..8228)
                     /label=AmpR promoter
     rep_origin      8510..9674
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
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 taccgccgcc
     1561 aattttaatc aaagtgggaa tattgctgat agctcattgt ccttcacttt cactaacagt
     1621 agcaacggtc cgaacctcat aacaactcaa acaaattctc aagcgctttc acaaccaatt
     1681 gcctcctcta acgttcatga taacttcatg aataatgaaa tcacggctag taaaattgat
     1741 gatggtaata attcaaaacc actgtcacct ggttggacgg accaaactgc gtataacgcg
     1801 tttggaatca ctacagggat gtttaatacc actacaatgg atgatgtata taactatcta
     1861 ttcgatgatg aagatacccc accaaaccca aaaaaagaga tctttaatac gactcactat
     1921 agggcgagcg ccgccatgga gtacccatac gacgtaccag attacgctca tatggccatg
     1981 gaggccagtg aattatcaac aagtttgtac aaaaaagctg aacgagaaac gtaaaatgat
     2041 ataaatatca atatattaaa ttagattttg cataaaaaac agactacata atactgtaaa
     2101 acacaacata tccagtcact atggcggccg cattaggcac cccaggcttt acactttatg
     2161 cttccggctc gtataatgtg tggattttga gttaggatcc ggcgagattt tcaggagcta
     2221 aggaagctaa aatggagaaa aaaatcactg gatataccac cgttgatata tcccaatggc
     2281 atcgtaaaga acattttgag gcatttcagt cagttgctca atgtacctat aaccagaccg
     2341 ttcagctgga tattacggcc tttttaaaga ccgtaaagaa aaataagcac aagttttatc
     2401 cggcctttat tcacattctt gcccgcctga tgaatgctca tccggaattc cgtatggcaa
     2461 tgaaagacgg tgagctggtg atatgggata gtgttcaccc ttgttacacc gttttccatg
     2521 agcaaactga aacgttttca tcgctctgga gtgaatacca cgacgatttc cggcagtttc
     2581 tacacatata ttcgcaagat gtggcgtgtt acggtgaaaa cctggcctat ttccctaaag
     2641 ggtttattga gaatatgttt ttcgtctcag ccaatccctg ggtgagtttc accagttttg
     2701 atttaaacgt ggccaatatg gacaacttct tcgcccccgt tttcaccatg ggcaaatatt
     2761 atacgcaagg cgacaaggtg ctgatgccgc tggcgattca ggttcatcat gccgtctgtg
     2821 atggcttcca tgtcggcaga atgcttaatg aattacaaca gtactgcgat gagtggcagg
     2881 gcggggcgta aagatctgga tccggcttac taaaagccag ataacagtat gcgtatttgc
     2941 gcgctgattt ttgcggtata agaatatata ctgatatgta tacccgaagt atgtcaaaaa
     3001 gaggtgtgct atgaagcagc gtattacagt gacagttgac agcgacagct atcagttgct
     3061 caaggcatat atgatgtcaa tatctccggt ctggtaagca caaccatgca gaatgaagcc
     3121 cgtcgtctgc gtgccgaacg ctggaaagcg gaaaatcagg aagggatggc tgaggtcgcc
     3181 cggtttattg aaatgaacgg ctcttttgct gacgagaaca gggactggtg aaatgcagtt
     3241 taaggtttac acctataaaa gagagagccg ttatcgtctg tttgtggatg tacagagtga
     3301 tattattgac acgcccgggc gacggatggt gatccccctg gccagtgcac gtctgctgtc
     3361 agataaagtc tcccgtgaac tttacccggt ggtgcatatc ggggatgaaa gctggcgcat
     3421 gatgaccacc gatatggcca gtgtgccggt ctccgttatc ggggaagaag tggctgatct
     3481 cagccaccgc gaaaatgaca tcaaaaacgc cattaacctg atgttctggg gaatataaat
     3541 gtcaggctcc cttatacaca gccagtctgc aggtcgacca tagtgactgg atatgttgtg
     3601 ttttacagta ttatgtagtc tgttttttat gcaaaatcta atttaatata ttgatattta
     3661 tatcatttta cgtttctcgt tcagctttct tgtacaaagt ggttgatgat ccatcgagct
     3721 cgagctgcag atgaatcgta gatactgaaa aaccccgcaa gttcacttca actgtgcatc
     3781 gtgcaccatc tcaatttctt tcatttatac atcgttttgc cttcttttat gtaactatac
     3841 tcctctaagt ttcaatcttg gccatgtaac ctctgatcta tagaattttt taaatgacta
     3901 gaattaatgc ccatcttttt tttggaccta aattcttcat gaaaatatat tacgagggct
     3961 tattcagaag ctttggactt cttcgccaga ggtttggtca agtctccaat caaggttgtc
     4021 ggcttgtcta ccttgccaga aatttacgaa aagatggaaa agggtcaaat cgttggtaga
     4081 tacgttgttg acacttctaa ataagcgaat ttcttatgat ttatgatttt tattattaaa
     4141 taagttataa aaaaaataag tgtatacaaa ttttaaagtg actcttaggt tttaaaacga
     4201 aaattcttat tcttgagtaa ctctttcctg taggtcaggt tgctttctca ggtatagcat
     4261 gaggtcgctc ttattgacca cacctctacc ggccggtcga aattccccta ccctatgaac
     4321 atattccatt ttgtaatttc gtgtcgtttc tattatgaat ttcatttata aagtttatgt
     4381 acaaatatca taaaaaaaga gaatcttttt aagcaaggat tttcttaact tcttcggcga
     4441 cagcatcacc gacttcggtg gtactgttgg aaccacctaa atcaccagtt ctgatacctg
     4501 catccaaaac ctttttaact gcatcttcaa tggccttacc ttcttcaggc aagttcaatg
     4561 acaatttcaa catcattgca gcagacaaga tagtggcgat agggttgacc ttattctttg
     4621 gcaaatctgg agcagaaccg tggcatggtt cgtacaaacc aaatgcggtg ttcttgtctg
     4681 gcaaagaggc caaggacgca gatggcaaca aacccaagga acctgggata acggaggctt
     4741 catcggagat gatatcacca aacatgttgc tggtgattat aataccattt aggtgggttg
     4801 ggttcttaac taggatcatg gcggcagaat caatcaattg atgttgaacc ttcaatgtag
     4861 gaaattcgtt cttgatggtt tcctccacag tttttctcca taatcttgaa gaggccaaaa
     4921 cattagcttt atccaaggac caaataggca atggtggctc atgttgtagg gccatgaaag
     4981 cggccattct tgtgattctt tgcacttctg gaacggtgta ttgttcacta tcccaagcga
     5041 caccatcacc atcgtcttcc tttctcttac caaagtaaat acctcccact aattctctga
     5101 caacaacgaa gtcagtacct ttagcaaatt gtggcttgat tggagataag tctaaaagag
     5161 agtcggatgc aaagttacat ggtcttaagt tggcgtacaa ttgaagttct ttacggattt
     5221 ttagtaaacc ttgttcaggt ctaacactac ctgtacccca tttaggacca cccacagcac
     5281 ctaacaaaac ggcatcaacc ttcttggagg cttccagcgc ctcatctgga agtgggacac
     5341 ctgtagcgtc gatagcagca ccaccaatta aatgattttc gaaatcgaac ttgacattgg
     5401 aacgaacatc agaaatagct ttaagaacct taatggcttc ggctgtgatt tcttgaccaa
     5461 cgtggtcacc tggcaaaacg acgatcttct taggggcaga cattagaatg gtatatcctt
     5521 gaaatatata tatatattgc tgaaatgtaa aaggtaagaa aagttagaaa gtaagacgat
     5581 tgctaaccac ctattggaaa aaacaatagg tccttaaata atattgtcaa cttcaagtat
     5641 tgtgatgcaa gcatttagtc atgaacgctt ctctattcta tatgaaaagc cggttccggc
     5701 gctctcacct ttcctttttc tcccaatttt tcagttgaaa aaggtatatg cgtcaggcga
     5761 cctctgaaat taacaaaaaa tttccagtca tcgaatttga ttctgtgcga tagcgcccct
     5821 gtgtgttctc gttatgttga ggaaaaaaat aatggttgct aagagattcg aactcttgca
     5881 tcttacgata cctgagtatt cccacagttg gggatctcga ctctagctag aggatcaatt
     5941 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     6001 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     6061 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctga
     6121 taacttcgta taatgtatgc tatacgaagt tattaggtct gaagaggagt ttacgtccag
     6181 ccaagctagc ttggctgcag gtcgagcggc cgcgatccgg aacccttaat ataacttcgt
     6241 ataatgtatg ctatacgaag ttatcagctg cattaatgaa tcggccaacg cgcggggaga
     6301 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     6361 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     6421 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     6481 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     6541 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     6601 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     6661 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     6721 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     6781 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     6841 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     6901 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     6961 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     7021 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     7081 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     7141 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     7201 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     7261 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     7321 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     7381 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     7441 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     7501 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     7561 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     7621 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     7681 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     7741 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     7801 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     7861 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     7921 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     7981 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     8041 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     8101 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     8161 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     8221 gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg
     8281 acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat
     8341 gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg
     8401 gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc
     8461 tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccataa cgcatttaag
     8521 cataaacacg cactatgccg ttcttctcat gtatatatat atacaggcaa cacgcagata
     8581 taggtgcgac gtgaacagtg agctgtatgt gcgcagctcg cgttgcattt tcggaagcgc
     8641 tcgttttcgg aaacgctttg aagttcctat tccgaagttc ctattctcta gctagaaagt
     8701 ataggaactt cagagcgctt ttgaaaacca aaagcgctct gaagacgcac tttcaaaaaa
     8761 ccaaaaacgc accggactgt aacgagctac taaaatattg cgaataccgc ttccacaaac
     8821 attgctcaaa agtatctctt tgctatatat ctctgtgcta tatccctata taacctaccc
     8881 atccaccttt cgctccttga acttgcatct aaactcgacc tctacatttt ttatgtttat
     8941 ctctagtatt actctttaga caaaaaaatt gtagtaagaa ctattcatag agtgaatcga
     9001 aaacaatacg aaaatgtaaa catttcctat acgtagtata tagagacaaa atagaagaaa
     9061 ccgttcataa ttttctgacc aatgaagaat catcaacgct atcactttct gttcacaaag
     9121 tatgcgcaat ccacatcggt atagaatata atcggggatg cctttatctt gaaaaaatgc
     9181 acccgcagct tcgctagtaa tcagtaaacg cgggaagtgg agtcaggctt tttttatgga
     9241 agagaaaata gacaccaaag tagccttctt ctaaccttaa cggacctaca gtgcaaaaag
     9301 ttatcaagag actgcattat agagcgcaca aaggagaaaa aaagtaatct aagatgcttt
     9361 gttagaaaaa tagcgctctc gggatgcatt tttgtagaac aaaaaagaag tatagattct
     9421 ttgttggtaa aatagcgctc tcgcgttgca tttctgttct gtaaaaatgc agctcagatt
     9481 ctttgtttga aaaattagcg ctctcgcgtt gcatttttgt tttacaaaaa tgaagcacag
     9541 attcttcgtt ggtaaaatag cgctttcgcg ttgcatttct gttctgtaaa aatgcagctc
     9601 agattctttg tttgaaaaat tagcgctctc gcgttgcatt tttgttctac aaaatgaagc
     9661 acagatgctt cgttgct
//
