LOCUS       40924_14395        8993 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Yeast vector pDEST-GBKT7, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8993)
  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  JOURNAL   Unpublished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 8993)
  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 8993)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8993)
  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            "Unpublished"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..8993
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        30..734
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             762..1202
                     /codon_start=1
                     /label=GAL4 DNA binding domain
                     /note="DNA binding domain of the GAL4 transcriptional
                     activator"
                     /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK
                     RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK
                     DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"
     promoter        1213..1231
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1251..1280
                     /codon_start=1
                     /label=Myc
                     /note="Myc (human c-Myc proto-oncogene) epitope tag"
                     /translation="EQKLISEEDL"
     protein_bind    1308..1432
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        1457..1487
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             1541..2197
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             2542..2844
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(2888..3012)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     terminator      3027..3074
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      3101..3288
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(3312..3328)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3336..3352)
                     /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(3360..3390)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3405..3426)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(3714..4302)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     polyA_signal    complement(4631..4678)
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal (Cole and Stacy, 1985)"
     CDS             complement(4914..5705)
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHDDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     promoter        complement(5706..5810)
                     /label=AmpR promoter
     rep_origin      complement(5837..7179)
                     /direction=LEFT
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     promoter        7438..7719
                     /label=TRP1 promoter
     CDS             7720..8391
                     /codon_start=1
                     /label=TRP1
                     /note="phosphoribosylanthranilate isomerase, required for 
                     tryptophan biosynthesis"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     rep_origin      complement(8497..8952)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 cggtgcgggc ctcttcgcta ttacgccaga tccttttgtt gtttccgggt gtacaatatg
       61 gacttcctct tttctggcaa ccaaacccat acatcgggat tcctataata ccttcgttgg
      121 tctccctaac atgtaggtgg cggaggggag atatacaata gaacagatac cagacaagac
      181 ataatgggct aaacaagact acaccaatta cactgcctca ttgatggtgg tacataacga
      241 actaatactg tagccctaga cttgatagcc atcatcatat cgaagtttca ctaccctttt
      301 tccatttgcc atctattgaa gtaataatag gcgcatgcaa cttcttttct ttttttttct
      361 tttctctctc ccccgttgtt gtctcaccat atccgcaatg acaaaaaaaa tgatggaaga
      421 cactaaagga aaaaattaac gacaaagaca gcaccaacag atgtcgttgt tccagagctg
      481 atgaggggta tctcgaagca cacgaaactt tttccttcct tcattcacgc acactactct
      541 ctaatgagca acggtatacg gccttccttc cagttacttg aatttgaaat aaaaaaagtt
      601 tgctgtcttg ctatcaagta taaatagacc tgcaattatt aatcttttgt ttcctcgtca
      661 ttgttctcgt tccctttctt ccttgtttct ttttctgcac aatatttcaa gctataccaa
      721 gcatacaatc aactccaagc ttgaagcaag cctcctgaaa gatgaagcta ctgtcttcta
      781 tcgaacaagc atgcgatatt tgccgactta aaaagctcaa gtgctccaaa gaaaaaccga
      841 agtgcgccaa gtgtctgaag aacaactggg agtgtcgcta ctctcccaaa accaaaaggt
      901 ctccgctgac tagggcacat ctgacagaag tggaatcaag gctagaaaga ctggaacagc
      961 tatttctact gatttttcct cgagaagacc ttgacatgat tttgaaaatg gattctttac
     1021 aggatataaa agcattgtta acaggattat ttgtacaaga taatgtgaat aaagatgccg
     1081 tcacagatag attggcttca gtggagactg atatgcctct aacattgaga cagcatagaa
     1141 taagtgcgac atcatcatcg gaagagagta gtaacaaagg tcaaagacag ttgactgtat
     1201 cgccggaatt tgtaatacga ctcactatag ggcgagccgc catcatggag gagcagaagc
     1261 tgatctcaga ggaggacctg catatggcca tggaggccga attatcaaca agtttgtaca
     1321 aaaaagctga acgagaaacg taaaatgata taaatatcaa tatattaaat tagattttgc
     1381 ataaaaaaca gactacataa tactgtaaaa cacaacatat ccagtcacta tggcggccgc
     1441 attaggcacc ccaggcttta cactttatgc ttccggctcg tataatgtgt ggattttgag
     1501 ttaggatccg gcgagatttt caggagctaa ggaagctaaa atggagaaaa aaatcactgg
     1561 atataccacc gttgatatat cccaatggca tcgtaaagaa cattttgagg catttcagtc
     1621 agttgctcaa tgtacctata accagaccgt tcagctggat attacggcct ttttaaagac
     1681 cgtaaagaaa aataagcaca agttttatcc ggcctttatt cacattcttg cccgcctgat
     1741 gaatgctcat ccggaattcc gtatggcaat gaaagacggt gagctggtga tatgggatag
     1801 tgttcaccct tgttacaccg ttttccatga gcaaactgaa acgttttcat cgctctggag
     1861 tgaataccac gacgatttcc ggcagtttct acacatatat tcgcaagatg tggcgtgtta
     1921 cggtgaaaac ctggcctatt tccctaaagg gtttattgag aatatgtttt tcgtctcagc
     1981 caatccctgg gtgagtttca ccagttttga tttaaacgtg gccaatatgg acaacttctt
     2041 cgcccccgtt ttcaccatgg gcaaatatta tacgcaaggc gacaaggtgc tgatgccgct
     2101 ggcgattcag gttcatcatg ccgtctgtga tggcttccat gtcggcagaa tgcttaatga
     2161 attacaacag tactgcgatg agtggcaggg cggggcgtaa agatctggat ccggcttact
     2221 aaaagccaga taacagtatg cgtatttgcg cgctgatttt tgcggtataa gaatatatac
     2281 tgatatgtat acccgaagta tgtcaaaaag aggtgtgcta tgaagcagcg tattacagtg
     2341 acagttgaca gcgacagcta tcagttgctc aaggcatata tgatgtcaat atctccggtc
     2401 tggtaagcac aaccatgcag aatgaagccc gtcgtctgcg tgccgaacgc tggaaagcgg
     2461 aaaatcagga agggatggct gaggtcgccc ggtttattga aatgaacggc tcttttgctg
     2521 acgagaacag ggactggtga aatgcagttt aaggtttaca cctataaaag agagagccgt
     2581 tatcgtctgt ttgtggatgt acagagtgat attattgaca cgcccgggcg acggatggtg
     2641 atccccctgg ccagtgcacg tctgctgtca gataaagtct cccgtgaact ttacccggtg
     2701 gtgcatatcg gggatgaaag ctggcgcatg atgaccaccg atatggccag tgtgccggtc
     2761 tccgttatcg gggaagaagt ggctgatctc agccaccgcg aaaatgacat caaaaacgcc
     2821 attaacctga tgttctgggg aatataaatg tcaggctccc ttatacacag ccagtctgca
     2881 ggtcgaccat agtgactgga tatgttgtgt tttacagtat tatgtagtct gttttttatg
     2941 caaaatctaa tttaatatat tgatatttat atcattttac gtttctcgtt cagctttctt
     3001 gtacaaagtg gttgatggcc gcataactag cataacccct tggggcctct aaacgggtct
     3061 tgaggggttt tttgcgcgct tgcagccaag ctaattccgg gcgaatttct tatgatttat
     3121 gatttttatt attaaataag ttataaaaaa aataagtgta tacaaatttt aaagtgactc
     3181 ttaggtttta aaacgaaaat tcttattctt gagtaactct ttcctgtagg tcaggttgct
     3241 ttctcaggta tagcatgagg tcgctcttat tgaccacacc tctaccggca tgcaagcttg
     3301 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     3361 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gaggtaactc
     3421 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     3481 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     3541 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     3601 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     3661 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     3721 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     3781 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     3841 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     3901 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     3961 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     4021 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     4081 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     4141 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     4201 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     4261 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     4321 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     4381 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     4441 taaagtatat atgagtaacc tgaggctatg gcagggcctg ccgccccgac gttggctgcg
     4501 agccctgggc cttcacccga acttgggggg tggggtgggg aaaaggaaga aacgcgggcg
     4561 tattggcccc aatggggtct cggtggggta tcgacagagt gccagccctg ggaccgaacc
     4621 ccgcgtttat gaacaaacga cccaacaccg tgcgttttat tctgtctttt tattgccgtc
     4681 atagcgcggg ttccttccgg tattgtctcc ttccgtgttt cagttagcct ccccctaggg
     4741 tgggcgaaga actccagcat gagatccccg cgctggagga tcatccagcc ggcgtcccgg
     4801 aaaacgattc cgaagcccaa cctttcatag aaggcggcgg tggaatcgaa atctcgtgat
     4861 ggcaggttgg gcgtcgcttg gtcggttcat ttcgaacccc agagtcccgc tcagaagaac
     4921 tcgtcaagaa ggcgatagaa ggcgatgcgc tgcgaatcgg gagcggcgat accgtaaagc
     4981 acgaggaagc ggtcagccca ttcgccgcca agctcttcag caatatcacg ggtagccaac
     5041 gctatgtcct gatagcggtc cgccacaccc agccggccac agtcgatgaa tccagaaaag
     5101 cggccatttt ccaccatgat attcggcaag caggcatcgt catgggtcac gacgagatcc
     5161 tcgccgtcgg gcatgctcgc cttgagcctg gcgaacagtt cggctggcgc gagcccctga
     5221 tgctcttcgt ccagatcatc ctgatcgaca agaccggctt ccatccgagt acgtgctcgc
     5281 tcgatgcgat gtttcgcttg gtggtcgaat gggcaggtag ccggatcaag cgtatgcagc
     5341 cgccgcattg catcagccat gatggatact ttctcggcag gagcaaggtg agatgacagg
     5401 agatcctgcc ccggcacttc gcccaatagc agccagtccc ttcccgcttc agtgacaacg
     5461 tcgagcacag ctgcgcaagg aacgcccgtc gtggccagcc acgatagccg cgctgcctcg
     5521 tcttgcagtt cattcagggc accggacagg tcggtcttga caaaaagaac cgggcgcccc
     5581 tgcgctgaca gccggaacac ggcggcatca gagcagccga ttgtctgttg tgcccagtca
     5641 tagccgaata gcctctccac ccaagcggcc ggagaacctg cgtgcaatcc atcttgttca
     5701 atcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     5761 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     5821 cgaaaagtgc cacctgaacg aagcatctgt gcttcatttt gtagaacaaa aatgcaacgc
     5881 gagagcgcta atttttcaaa caaagaatct gagctgcatt tttacagaac agaaatgcaa
     5941 cgcgaaagcg ctattttacc aacgaagaat ctgtgcttca tttttgtaaa acaaaaatgc
     6001 aacgcgagag cgctaatttt tcaaacaaag aatctgagct gcatttttac agaacagaaa
     6061 tgcaacgcga gagcgctatt ttaccaacaa agaatctata cttctttttt gttctacaaa
     6121 aatgcatccc gagagcgcta tttttctaac aaagcatctt agattacttt ttttctcctt
     6181 tgtgcgctct ataatgcagt ctcttgataa ctttttgcac tgtaggtccg ttaaggttag
     6241 aagaaggcta ctttggtgtc tattttctct tccataaaaa aagcctgact ccacttcccg
     6301 cgtttactga ttactagcga agctgcgggt gcattttttc aagataaagg catccccgat
     6361 tatattctat accgatgtgg attgcgcata ctttgtgaac agaaagtgat agcgttgatg
     6421 attcttcatt ggtcagaaaa ttatgaacgg tttcttctat tttgtctcta tatactacgt
     6481 ataggaaatg tttacatttt cgtattgttt tcgattcact ctatgaatag ttcttactac
     6541 aatttttttg tctaaagagt aatactagag ataaacataa aaaatgtaga ggtcgagttt
     6601 agatgcaagt tcaaggagcg aaaggtggat gggtaggtta tatagggata tagcacagag
     6661 atatatagca aagagatact tttgagcaat gtttgtggaa gcggtattcg caatatttta
     6721 gtagctcgtt acagtccggt gcgtttttgg ttttttgaaa gtgcgtcttc agagcgcttt
     6781 tggttttcaa aagcgctctg aagttcctat actttctaga gaataggaac ttcggaatag
     6841 gaacttcaaa gcgtttccga aaacgagcgc ttccgaaaat gcaacgcgag ctgcgcacat
     6901 acagctcact gttcacgtcg cacctatatc tgcgtgttgc ctgtatatat atatacatga
     6961 gaagaacggc atagtgcgtg tttatgctta aatgcgtact tatatgcgtc tatttatgta
     7021 ggatgaaagg tagtctagta cctcctgtga tattatccca ttccatgcgg ggtatcgtat
     7081 gcttccttca gcactaccct ttagctgttc tatatgctgc cactcctcaa ttggattagt
     7141 ctcatccttc aatgctatca tttcctttga tattggatca tattaagaaa ccattattat
     7201 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg
     7261 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     7321 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     7381 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atagatcaac
     7441 gacattacta tatatataat ataggaagca tttaatagaa cagcatcgta atatatgtgt
     7501 actttgcagt tatgacgcca gatggcagta gtggaagata ttctttattg aaaaatagct
     7561 tgtcacctta cgtacaatct tgatccggag cttttctttt tttgccgatt aagaattaat
     7621 tcggtcgaaa aaagaaaagg agagggccaa gagggagggc attggtgact attgagcacg
     7681 tgagtatacg tgattaagca cacaaaggca gcttggagta tgtctgttat taatttcaca
     7741 ggtagttctg gtccattggt gaaagtttgc ggcttgcaga gcacagaggc cgcagaatgt
     7801 gctctagatt ccgatgctga cttgctgggt attatatgtg tgcccaatag aaagagaaca
     7861 attgacccgg ttattgcaag gaaaatttca agtcttgtaa aagcatataa aaatagttca
     7921 ggcactccga aatacttggt tggcgtgttt cgtaatcaac ctaaggagga tgttttggct
     7981 ctggtcaatg attacggcat tgatatcgtc caactgcatg gagatgagtc gtggcaagaa
     8041 taccaagagt tcctcggttt gccagttatt aaaagactcg tatttccaaa agactgcaac
     8101 atactactca gtgcagcttc acagaaacct cattcgttta ttcccttgtt tgattcagaa
     8161 gcaggtggga caggtgaact tttggattgg aactcgattt ctgactgggt tggaaggcaa
     8221 gagagccccg aaagcttaca ttttatgtta gctggtggac tgacgccaga aaatgttggt
     8281 gatgcgctta gattaaatgg cgttattggt gttgatgtaa gcggaggtgt ggagacaaat
     8341 ggtgtaaaag actctaacaa aatagcaaat ttcgtcaaaa atgctaagaa ataggttatt
     8401 actgagtagt atttatttaa gtattgtttg tgcacttgcc gatctatgcg gtgtgaaata
     8461 ccgcacagat gcgtaaggag aaaataccgc atcaggaaat tgtaaacgtt aatattttgt
     8521 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     8581 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     8641 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     8701 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     8761 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     8821 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     8881 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     8941 tacagggcgc gtccattcgc cattcaggct gcgcaactgt tgggaagggc gat
//
