LOCUS       40924_4904        9534 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Yeast expression vector pAS3, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9534)
  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  JOURNAL   Unpublished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 9534)
  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 9534)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 9534)
  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            "Unpublished"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..9534
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(2..1344)
                     /direction=LEFT
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     promoter        1603..1883
                     /label=TRP1 promoter
     promoter        3101..3476
                     /label=ADE2 promoter
     CDS             3477..5189
                     /codon_start=1
                     /label=ADE2
                     /note="phosphoribosylaminoimidazole carboxylase, required
                     for adenine biosynthesis"
                     /translation="MDSRTVGILGGGQLGRMIVEAANRLNIKTVILDAENSPAKQISNS
                     NDHVNGSFSNPLDIEKLAEKCDVLTIEIEHVDVPTLKNLQVKHPKLKIYPSPETIRLIQ
                     DKYIQKEHLIKNGIAVTQSVPVEQASETSLLNVGRDLGFPFVLKSRTLAYDGRGNFVVK
                     NKEMIPEALEVLKDRPLYAEKWAPFTKELAVMIVRSVNGLVFSYPIVETIHKDNICDLC
                     YAPARVPDSVQLKAKLLAENAIKSFPGCGIFGVEMFYLETGELLINEIAPRPHNSGHYT
                     IDACVTSQFEAHLRSILDLPMPKNFTSFSTITTNAIMLNVLGDKHTKDKELETCERALA
                     TPGSSVYLYGKESRPNRKVGHINIIASSMAECEQRLNYITGRTDIPIKISVAQKLDLEA
                     MVKPLVGIIMGSDSDLPVMSAACAVLKDFGVPFEVTIVSAHRTPHRMSAYAISASKRGI
                     KTIIAGAGGAAHLPGMVAAMTPLPVIGVPVKGSCLDGVDSLHSIVQMPRGVPVATVAIN
                     NSTNAALLAVRLLGAYDSSYTTKMEQFLLKQEEEVLVKAQKLETVGYEAYLENK"
     CDS             complement(5491..5502)
                     /codon_start=1
                     /label=WELQut site
                     /note="WELQut protease recognition and cleavage site"
                     /translation="WELQ"
     promoter        5890..6594
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             6622..7062
                     /codon_start=1
                     /label=GAL4 DNA binding domain
                     /note="DNA binding domain of the GAL4 transcriptional
                     activator"
                     /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK
                     RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK
                     DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"
     CDS             7078..7104
                     /codon_start=1
                     /label=HA
                     /note="HA (human influenza hemagglutinin) epitope tag"
                     /translation="YPYDVPDYA"
     terminator      7172..7359
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(7383..7399)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(7407..7423)
                     /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(7431..7461)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(7476..7497)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(7785..8373)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(8547..9404)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(9405..9509)
                     /label=AmpR promoter
ORIGIN
        1 gaacgaagca tctgtgcttc attttgtaga acaaaaatgc aacgcgagag cgctaatttt
       61 tcaaacaaag aatctgagct gcatttttac agaacagaaa tgcaacgcga aagcgctatt
      121 ttaccaacga agaatctgtg cttcattttt gtaaaacaaa aatgcaacgc gagagcgcta
      181 atttttcaaa caaagaatct gagctgcatt tttacagaac agaaatgcaa cgcgagagcg
      241 ctattttacc aacaaagaat ctatacttct tttttgttct acaaaaatgc atcccgagag
      301 cgctattttt ctaacaaagc atcttagatt actttttttc tcctttgtgc gctctataat
      361 gcagtctctt gataactttt tgcactgtag gtccgttaag gttagaagaa ggctactttg
      421 gtgtctattt tctcttccat aaaaaaagcc tgactccact tcccgcgttt actgattact
      481 agcgaagctg cgggtgcatt ttttcaagat aaaggcatcc ccgattatat tctataccga
      541 tgtggattgc gcatactttg tgaacagaaa gtgatagcgt tgatgattct tcattggtca
      601 gaaaattatg aacggtttct tctattttgt ctctatatac tacgtatagg aaatgtttac
      661 attttcgtat tgttttcgat tcactctatg aatagttctt actacaattt ttttgtctaa
      721 agagtaatac tagagataaa cataaaaaat gtagaggtcg agtttagatg caagttcaag
      781 gagcgaaagg tggatgggta ggttatatag ggatatagca cagagatata tagcaaagag
      841 atacttttga gcaatgtttg tggaagcggt attcgcaata ttttagtagc tcgttacagt
      901 ccggtgcgtt tttggttttt tgaaagtgcg tcttcagagc gcttttggtt ttcaaaagcg
      961 ctctgaagtt cctatacttt ctagagaata ggaacttcgg aataggaact tcaaagcgtt
     1021 tccgaaaacg agcgcttccg aaaatgcaac gcgagctgcg cacatacagc tcactgttca
     1081 cgtcgcacct atatctgcgt gttgcctgta tatatatata catgagaaga acggcatagt
     1141 gcgtgtttat gcttaaatgc gtacttatat gcgtctattt atgtaggatg aaaggtagtc
     1201 tagtacctcc tgtgatatta tcccattcca tgcggggtat cgtatgcttc cttcagcact
     1261 accctttagc tgttctatat gctgccactc ctcaattgga ttagtctcat ccttcaatgc
     1321 tatcatttcc tttgatattg gatcatatta agaaaccatt attatcatga cattaaccta
     1381 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa
     1441 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     1501 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta
     1561 tgcggcatca gagcagattg tactgagagt gcaccataga tcaacgacat tactatatat
     1621 ataatatagg aagcatttaa tagacagcat cgtaatatat gtgtactttg cagttatgac
     1681 gccagatggc agtagtggaa gatattcttt attgaaaaat agcttgtcac cttacgtaca
     1741 atcttgatcc ggagcttttc tttttttgcc gattaagaat taattcggtc gaaaaaagaa
     1801 aaggagaggg ccaagaggga gggcattggt gactattgag cacgtgagta tacgtgatta
     1861 agcacacaaa ggcagcttgg agtatgtctg ttattaattt cacaggtagt tctggtccat
     1921 tggtgaaagt ttgcggcttg cagagcacag aggccgcaga atgtgctcta gattccgatg
     1981 ctgacttgct gggtattata tgtgtgccca atagaaagag aacaattgac ccggttattg
     2041 caaggaaaat ttcaagtctt gtaaaagcat ataaaaatag ttcaggcact ccgaaatact
     2101 tggttggcgt gtttcgtaat caacctaagg aggatgtttt ggctctggtc aatgattacg
     2161 gcattgatnc cacggtgtta tagtatcagc aatagatctt tttactcaaa tttacagttt
     2221 tgatatcttg gcgtttttcc tcacatttat tacaaaagaa agtgggcagg atttgccctt
     2281 cacaaaatct ctgtccagga gaagaacgta tttgaccaga attgctcaat cttcttcatt
     2341 gagacagttt ttacaactga gtacacgccc taatataagg attctgggag gtaatgggca
     2401 gggtactgga aattccaatt atcccgaatt tacaaacgca tcctcagtca tctcacccaa
     2461 gctaagggct tctcctctac tagaaagaag ggcgtcaaaa atctgctatg ctattacgaa
     2521 attactctat aggttagaat gtcatcccat aggtatggcc cttttaaaga agtatctcca
     2581 caatcaattg cgagaagcct atctagaatc aaaacgacac tttatttcca aaaagggaga
     2641 ttcgaccaac acttcctcta ccattgcatc atcatctttc gctggcgcat ctgttcctct
     2701 atcttcaaac gaatcaggaa tgctaaacgg cttgaagcaa attaacgaac aacaagaatc
     2761 tacattagaa accactcaaa aggaagacta gtaacgccgt atcgtgatta acgtattaca
     2821 taagttacag gattcatgct tatgggttag ctatttcgcc caatgtgtcc atctgacatt
     2881 actattttgc attttaattt aattagaact tgactagcgc actaccagta tatcatctca
     2941 tttccgtaaa taccaaatgt attatatatt gaaagctttt gaccaggtta ttataaaaga
     3001 aacttcatgc tcgaaaaaga tcatttcgaa aagttgccta gtttcatgaa attttaaagc
     3061 agtttatata aattttacct tttgatgcgg aattgacttt ttcttgaata atacataact
     3121 tttcttaaaa gaatcaaaga cagataaaat ttaagagata ttaaatatta gtgagaagcc
     3181 gagaattttg taacaccaac ataacactga catctttaac aacttttaat tatgatacat
     3241 ttcttacgtc atgattgatt attacagcta tgctgacaaa tgactcttgt tgcatggcta
     3301 cgaaccgggt aatactaagt gattgactct tgctgacctt ttattaagaa ctaaatggac
     3361 aatattatgg agcatttcat gtataaattg gtgcgtaaaa tcgttggatc tctcttctaa
     3421 gtacatccta ctataacaat caagaaaaac aagaaaatcg gacaaaacaa tcaagtatgg
     3481 attctagaac agttggtata ttaggagggg gacaattggg acgtatgatt gttgaggcag
     3541 caaacaggct caacattaag acggtaatac tagatgctga aaattctcct gccaaacaaa
     3601 taagcaactc caatgaccac gttaatggct ccttttccaa tcctcttgat atcgaaaaac
     3661 tagctgaaaa atgtgatgtg ctaacgattg agattgagca tgttgatgtt cctacactaa
     3721 agaatcttca agtaaaacat cccaaattaa aaatttaccc ttctccagaa acaatcagat
     3781 tgatacaaga caaatatatt caaaaagagc atttaatcaa aaatggtata gcagttaccc
     3841 aaagtgttcc tgtggaacaa gccagtgaga cgtccctatt gaatgttgga agagatttgg
     3901 gttttccatt cgtcttgaag tcgaggactt tggcatacga tggaagaggt aacttcgttg
     3961 taaagaataa ggaaatgatt ccggaagctt tggaagtact gaaggatcgt cctttgtacg
     4021 ccgaaaaatg ggcaccattt actaaagaat tagcagtcat gattgtgaga tctgttaacg
     4081 gtttagtgtt ttcttaccca attgtagaga ctatccacaa ggacaatatt tgtgacttat
     4141 gttatgcgcc tgctagagtt ccggactccg ttcaacttaa ggcgaagttg ttggcagaaa
     4201 atgcaatcaa atcttttccc ggttgtggta tatttggtgt ggaaatgttc tatttagaaa
     4261 caggggaatt gcttattaac gaaattgccc caaggcctca caactctgga cattatacca
     4321 ttgatgcttg cgtcacttct caatttgaag ctcatttgag atcaatattg gatttgccaa
     4381 tgccaaagaa tttcacatct ttctccacca ttacaacgaa cgccattatg ctaaatgttc
     4441 ttggagacaa acatacaaaa gataaagagc tagaaacttg cgaaagagca ttggcgactc
     4501 caggttcctc agtgtactta tatggaaaag agtctagacc taacagaaaa gtaggtcaca
     4561 taaatattat tgcctccagt atggcggaat gtgaacaaag gctgaactac attacaggta
     4621 gaactgatat tccaatcaaa atctctgtcg ctcaaaagtt ggacttggaa gcaatggtca
     4681 aaccattggt tggaatcatc atgggatcag actctgactt gccggtaatg tctgccgcat
     4741 gtgcggtttt aaaagatttt ggcgttccat ttgaagtgac aatagtctct gctcatagaa
     4801 ctccacatag gatgtcagca tatgctattt ccgcaagcaa gcgtggaatt aaaacaatta
     4861 tcgctggagc tggtggggct gctcacttgc caggtatggt ggctgcaatg acaccacttc
     4921 ctgtcatcgg tgtgcccgta aaaggttctt gtctagatgg agtagattct ttacattcaa
     4981 ttgtgcaaat gcctagaggt gttccagtag ctaccgtcgc tattaataat agtacgaacg
     5041 ctgcgctgtt ggctgtcaga ctgcttggcg cttatgattc aagttataca acgaaaatgg
     5101 aacagttttt attaaagcaa gaagaagaag ttcttgtcaa agcacaaaag ttagaaactg
     5161 tcggttacga agcttatcta gaaaacaagt aatatataag tttattgata tacttgtaca
     5221 gcaaataatt ataaaatgat atacctattt tttaggcttt gttatgatta catcaaatgt
     5281 ggacttcata catagaaatc aacgcttaca ggtgtccttt tttaagaatt tcatacataa
     5341 gatcacttat tatacataca tacatatcca gtaacaagaa gcaaggaata attacctgct
     5401 taagtctgcg attaaaaaaa taacgtttcg atacagttca tataaggcgg ctcaatgcag
     5461 aaccgaggat agcgctacgt caggatatct ttgtagttcc caaatataaa tgcgacaata
     5521 tagtttcttt ctttcatatc aataatatcc ttttctccac tgaaatcacg aatcaaacct
     5581 ggagcaaaaa ctaaagccaa attataaagc gtcattcgat tccagtgact gtaccgtgta
     5641 accttttcta tatgttcact cagtactctt aacaccctat aatgttccct tggaaggtct
     5701 tccaatatgt tttttaaagc gctcttgctc gacatataag tgtccgaatt ctttgcttct
     5761 aaggacaact ttcctccaac aaatggcaag ttttccatca tttttttaga tttaactaac
     5821 ctcatcaacg gctcgtatat ttgaaaggta aagatagggt tggggagctt tcttaagtat
     5881 cgcttcaana tccttttgtt gtttccgggt gtacaatatg gacttcctct tttctggcaa
     5941 ccaaacccat acatcgggat tcctataata ccttcgttgg tctccctaac atgtaggtgg
     6001 cggaggggag atatacaata gaacagatac cagacaagac ataatgggct aaacaagact
     6061 acaccaatta cactgcctca ttgatggtgg tacataacga actaatactg tagccctaga
     6121 cttgatagcc atcatcatat cgaagtttca ctaccctttt tccatttgcc atctattgaa
     6181 gtaataatag gcgcatgcaa cttcttttct ttttttttct tttctctctc ccccgttgtt
     6241 gtctcaccat atccgcaatg acaaaaaaaa tgatggaaga cactaaagga aaaaattaac
     6301 gacaaagaca gcaccaacag atgtcgttgt tccagagctg atgaggggta tctcgaagca
     6361 cacgaaactt tttccttcct tcattcacgc acactactct ctaatgagca acggtatacg
     6421 gccttccttc cagttacttg aatttgaaat aaaaaaagtt tgccgctttg ctatcaagta
     6481 taaatagacc tgcaattatt aatcttttgt ttcctcgtca ttgttctcgt tccctttctt
     6541 ccttgtttct ttttctgcac aatatttcaa gctataccaa gcatacaatc aactccaagc
     6601 ttgaagcaag cctcctgaaa gatgaagcta ctgtcttcta tcgaacaagc atgcgatatt
     6661 tgccgactta aaaagctcaa gtgctccaaa gaaaaaccga agtgcgccaa gtgtctgaag
     6721 aacaactggg agtgtcgcta ctctcccaaa accaaaaggt ctccgctgac tagggcacat
     6781 ctgacagaag tggaatcaag gctagaaaga ctggaacagc tatttctact gatttttcct
     6841 cgagaagacc ttgacatgat tttgaaaatg gattctttac aggatataaa agcattgtta
     6901 acaggattat ttgtacaaga taatgtgaat aaagatgccg tcacagatag attggcttca
     6961 gtggagactg atatgcctct aacattgaga cagcatagaa taagtgcgac atcatcatcg
     7021 gaagagagta gtaacaaagg tcaaagacag ttgactgtat cgccggaatt catggcttac
     7081 ccatacgatg ttccagatta cgctagcttg ggtggtcata tggccatgga ggccccgggg
     7141 atccgtcgac ctgcagccaa gctaattccg ggcgaatttc ttatgattta tgatttttat
     7201 tattaaataa gttataaaaa aaataagtgt atacaaattt taaagtgact cttaggtttt
     7261 aaaacgaaaa ttcttgttct tgagtaactc tttcctgtag gtcaggttgc tttctcaggt
     7321 atagcatgag gtcgctctta ttgaccacac ctctaccggc atgcaagctt ggcgtaatca
     7381 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     7441 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgaggtaact cacattaatt
     7501 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct ggattaatga
     7561 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     7621 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     7681 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     7741 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     7801 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     7861 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     7921 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     7981 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     8041 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     8101 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     8161 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     8221 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     8281 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     8341 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     8401 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     8461 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     8521 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     8581 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     8641 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     8701 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     8761 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     8821 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     8881 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     8941 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     9001 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     9061 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     9121 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     9181 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     9241 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     9301 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     9361 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     9421 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     9481 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acct
//
