LOCUS       40924_31015        8330 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Yeast two-hybrid vector pMK500-TEV, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8330)
  AUTHORS   Guo D, Hazbun TR, Xu XJ, Ng SL, Fields S, Kuo MH.
  TITLE     A tethered catalysis, two-hybrid system to identify protein-protein 
            interactions requiring post-translational modifications
  JOURNAL   Nat. Biotechnol. 22 (7), 888-892 (2004)
  PUBMED    15208639
REFERENCE   2  (bases 1 to 8330)
  AUTHORS   Kuo M-H., Xu X-J., Ng S-L.
  TITLE     Direct Submission
  JOURNAL   Submitted (08-JUN-2004) Biochem. Mol. Biol., Michigan State 
            University, 309 BCH Building, East Lansing, MI 48824, USA
REFERENCE   3  (bases 1 to 8330)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8330)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat. 
            Biotechnol."; date: "2004"; volume: "22"; issue: "7"; pages: 
            "888-892"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (08-JUN-2004) Biochem. Mol. Biol., Michigan State University, 309 
            BCH Building, East Lansing, MI 48824, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8330
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             434..874
                     /label=GAL4 DNA binding domain
                     /note="DNA binding domain of the GAL4 transcriptional
                     activator"
     CDS             932..952
                     /label=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
     misc_feature    962..1024
                     /note="trimeric carboxyl terminal domain repeat of RNA 
                     polymerase II"
     CDS             1040..1957
                     /gene="KIN28"
                     /label=KIN28
                     /note="Serine/threonine-protein kinase KIN28 from
                     Saccharomyces cerevisiae (strain ATCC 204508 / S288c). 
                     Accession#: P06242"
     CDS             1991..2017
                     /label=HA
                     /note="HA (human influenza hemagglutinin) epitope tag"
     CDS             2021..2047
                     /label=HA
                     /note="HA (human influenza hemagglutinin) epitope tag"
     CDS             2054..2080
                     /label=HA
                     /note="HA (human influenza hemagglutinin) epitope tag"
     terminator      2519..2706
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae
                     alcohol dehydrogenase 1 (ADH1) gene"
     misc_feature    complement(3538..4700)
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN4 centromere fused to the
                     autonomously replicating sequence ARS1/ARS416"
     promoter        complement(5214..5315)
                     /label=TRP1 promoter
     promoter        5421..5525
                     /label=AmpR promoter
     CDS             5526..6383
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      6557..7145
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gtggtacata acgaactaat actgtagccc tagacttgat agccatcatc atatcgaagt
       61 ttcactaccc tttttccatt tgccatctat tgaagtaata ataggcgcat gcaacttctt
      121 ttcttttttt ttcttttctc tctcccccgt tgttgtctca ccatatccgc aatgacaaaa
      181 aaatgatgga agacactaaa ggaaaaaatt aacgacaaag acagcaccaa cagatgtcgt
      241 tgttccagag ctgatgaggg gtatctcgaa gcacacgaaa ctttttcctt ccttcattga
      301 cctgcaatta ttaatctttt gtttcctcgt cattgttctc gttccctttc ttccttgttt
      361 ctttttctgc acaatatttc aagctatacc aagcatacaa tcaactccaa gcttgaagca
      421 agcctcctga aagatgaagc tactgtcttc tatcgaacaa gcatgcgata tttgccgact
      481 taaaaagctc aagtgctcca aagaaaaacc gaagtgcgcc aagtgtctga agaacaactg
      541 ggagtgtcgc tactctccca aaaccaaaag gtctccgctg actagggcac atctgacaga
      601 agtggaatca aggctagaaa gactggaaca gctatttcta ctgatttttc ctcgagaaga
      661 ccttgacatg attttgaaaa tggattcttt acaggatata aaagcattgt taacaggatt
      721 atttgtacaa gataatgtga ataaagatgc cgtcacagat agattggctt cagtggagac
      781 tgatatgcct ctaacattga gacagcatag aataagtgcg acatcatcat cggaagagag
      841 tagtaacaaa ggtcaaagac agttgactgt atcgaactat ctattcgatg atgaagatac
      901 cccaccaaac ccaaaaaaag agatcgaatt ggagaatttg tattttcagg gtgaattcca
      961 gtattcgcca actagtcctt cgtattcgcc aactagtcct tcgtattcgc caactagtcc
     1021 ttcgctgacc accatggcaa tgaaagtgaa tatggagtac acaaaggaaa agaaagttgg
     1081 tgagggtact tatgcggttg tttacttggg ttgtcaacac tctactggaa gaaagattgc
     1141 tattaaggag atcaaaacat ccgaatttaa agatggttta gatatgtcag ctatccgtga
     1201 agttaagtac ctccaagaaa tgcagcatcc gaacgtcata gaactaatag acatatttat
     1261 ggcttatgat aatttaaatc tcgttctgga gttcctacca actgatctag aggtggtaat
     1321 aaaagacaaa tcaatactgt ttacaccagc agatattaag gcatggatgc ttatgacttt
     1381 gaggggcgtg tatcattgcc acagaaattt cattttgcac agggatctga aaccaaacaa
     1441 tttattattt tcacctgatg gccagataaa agtagcagat ttcggtctag caagggcgat
     1501 accggcccca catgagatac tgacaagtaa cgtcgtaaca agatggtata gagcgccaga
     1561 attgttgttt ggagctaaac attacacatc ggctattgat atctggtcag taggcgttat
     1621 attcgcggaa ttaatgctaa ggatacctta tttaccagga cagaatgatg tcgatcaaat
     1681 ggaagtaacg ttcagggcct tagggacacc tacagataga gattggcccg aagtttcttc
     1741 ctttatgacg tataacaagt tacaaatata tccgccccct tcaagagatg aattgaggaa
     1801 aaggttcatt gctgctagcg aatacgcctt agattttatg tgtggaatgc taacgatgaa
     1861 cccacaaaag aggtggaccg ctgttcagtg tttagaaagt gattatttca aagaattacc
     1921 accaccaagt gacccgtctt caataaaaat acgtaacgtc atggcaattc ccggtggcgg
     1981 ccgcatcttt tacccatacg atgttcctga ctatgcgggc tatccctatg acgtcccgga
     2041 ctatgcagga tcctatccat atgacgttcc agattacgct gctcagtgcg gccgctctag
     2101 ctagaactag tggatccccc gataccgtcg acctgcagag atctatgaat cgtagatact
     2161 gaaaaacccc gcaagttcac ttcaactgtg catcgtgcac catctcaatt tctttcattt
     2221 atacatcgtt ttgccttctt ttatgtaact atactcctct aagtttcaat cttggccatg
     2281 taacctctga tctatagaat tttttaaatg actagaatta atgcccatct tttttttgga
     2341 cctaaattct tcatgaaaat atattacgag ggcttattca gaagctttgg acttcttcgc
     2401 cagaggtttg gtcaagtctc caatcaaggt tgtcggcttg tctaccttgc cagaaattta
     2461 cgaaaagatg gaaaagggtc aaatcgttgg tagatacgtt gttgacactt ctaaataagc
     2521 gaatttctta tgatttatga tttttattat taaataagtt ataaaaaaaa taagtgtata
     2581 caaattttaa agtgactctt aggttttaaa acgaaaattc ttattcttga gtaactcttt
     2641 cctgtaggtc aggttgcttt ctcaggtata gcatgaggtc gctcttattg accacacctc
     2701 taccggcatg ccgagcaaat gcctgcaaat cgctccccat ttcacccaat tgtagatatg
     2761 ctaactccag caatgagttg atgaatctcg gtgtgtattt tatgtcctca gaggacaaca
     2821 cctgttgtaa tcgttcttcc acacggatcc tggcgtaata gcgaagaggc ccgcaccgat
     2881 cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg gtattttctc
     2941 cttacgcatc tgtgcggtat ttcacaccgc atatatcgct gggccattct catgaagaat
     3001 atcttgaatt tattgtcata ttactagttg gtgtggaagt ccatatatcg gtgatcaata
     3061 tagtggttga catgctggct agtcaacatt gagccttttg atcatgcaaa tatattacgg
     3121 tattttacaa tcaaatatca aacttaacta ttgactttat aacttattta ggtggtaaca
     3181 ttcttataaa aaagaaaaaa attactgcaa aacagtacta gcttttaact tgtatcctag
     3241 gttatctatg ctgtctcacc atagagaata ttacctattt cagaatgtat gtccatgatt
     3301 cgccgggtaa atacatataa tacacaaatc tggcttaata aagtctataa tatatctcat
     3361 aaagaagtgc taaattggct agtgctatat atttttaaga aaatttcttt tgactaagtc
     3421 catatcgact ttgtaaaagt tcactttagc atacatatat tacacgagcc agaaattgta
     3481 acttttgcct aaaatcacaa attgcaaaat ttaattgctt gcaaaaggtc acatgcttat
     3541 aatcaacttt tttaaaaatt taaaatactt ttttattttt tatttttaaa cataaatgaa
     3601 ataatttatt tattgtttat gattaccgaa acataaaacc tgctcaagaa aaagaaactg
     3661 ttttgtcctt ggaaaaaaag cactacctag gagcggccaa aatgccgagg ctttcatagc
     3721 ttaaactctt tacagaaaat aggcattata gatcagttcg agttttctta ttcttccttc
     3781 cggttttatc gtcacagttt tacagtaaat aagtatcacc tcttagagtt cgatgataag
     3841 ctgtcaaaca tgagaattaa ttccacatgt taaaatagtg aaggagcatg ttcggcacac
     3901 agtggaccga acgtggggta agtgcactag ggtccggtta aacggatctc gcattgatga
     3961 ggcaacgcta attatcaaca tatagattgt tatctatctg catgaacacg aaatctttac
     4021 ttgacgactt gaggctgatg gtgtttatgc aaagaaacca ctgtgtttaa tatgtgtcac
     4081 tgtttgatat tactgtcagc gtagaagata atagtaaaag cggttaataa gtgtatttga
     4141 gataagtgtg ataaagtttt tacagcgaaa agacgataaa tacaagaaaa tgattacgag
     4201 gatacggaga gaggtatgta catgtgtatt tatatactaa gctgccggcg gttgtttgca
     4261 agaccgagaa aaggctagca agaatcgggt cattgtagcg tatgcgcctg tgaacattct
     4321 cttcaacaag tttgattcca ttgcggtgaa atggtaaaag tcaaccccct gcgatgtata
     4381 ttttcctgta caatcaatca aaaagccaaa tgatttagca ttatctttac atcttgttat
     4441 tttacagatt ttatgtttag atcttttatg cttgcttttc aaaaggcttg caggcaagtg
     4501 cacaaacaat acttaaataa atactactca gtaataacct atttcttagc atttttgacg
     4561 aaatttgcta ttttgttaga gtcttttaca ccatttgtct ccacacctcc gcttacatca
     4621 acaccaataa cgccatttaa tctaagcgca tcaccaacat tttctggcgt cagtccacca
     4681 gctaacataa aatgtaagct ctcggggctc tcttgccttc caacccagtc agaaatcgag
     4741 ttccaatcca aaagttcacc tgtcccacct gcttctgaat caaacaaggg aataaacgaa
     4801 tgaggtttct gtgaagctgc actgagtagt atgttgcagt cttttggaaa tacgagtctt
     4861 ttaataactg gcaaaccgag gaactcttgg tattcttgcc acgactcatc tccatgcagt
     4921 tggacgatat caatgccgta atcattgacc agagccaaaa catcctcctt aggttgatta
     4981 cgaaacacgc caaccaagta tttcggagtg cctgaactat ttttatatgc ttttacaaga
     5041 cttgaaattt tccttgcaat aaccgggtca attgttctct ttctattggg cacacatata
     5101 atacccagca agtcagcatc ggaatctagt gcacattctg cggcctctgt gctctgcaag
     5161 ccgcaaactt tcaccaatgg accagaacta cctgtgaaat taataacaga catactccaa
     5221 gctgcctttg tgtgcttaat cacgtatact cacgtgctca atagtcacca atgccctccc
     5281 tcttggccct ctccttttct tttttcgacc gaattaattc ttgaagacga aagggcctcg
     5341 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     5401 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     5461 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     5521 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     5581 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     5641 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     5701 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     5761 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     5821 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     5881 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     5941 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     6001 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     6061 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     6121 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     6181 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     6241 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     6301 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     6361 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     6421 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     6481 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     6541 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     6601 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     6661 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     6721 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     6781 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     6841 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     6901 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     6961 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     7021 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     7081 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     7141 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     7201 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     7261 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     7321 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     7381 ggatccggga tcgaagaaat gatggtaaat gaaataggaa atcaaggagc atgaaggcaa
     7441 aagacaaata taagggtcga acgaaaaata aagtgaaaag tgttgatatg atgtatttgg
     7501 ctttgcggcg ccgaaaaaac gagtttacgc aattgcacaa tcatgctgac tctgtggcgg
     7561 acccgcgctc ttgccggccc ggcgataacg ctgggcgtga ggctgtgccc ggcggagttt
     7621 tttgcgcctg cattttccaa ggtttaccct gcgctaaggg gcgagattgg agaagcaata
     7681 agaatgccgg ttggggttgc gatgatgacg accacgacaa ctggtgtcat tatttaagtt
     7741 gccgaaagaa cctgagtgca tttgcaacat gagtatacta gaagaatgag ccaagacttg
     7801 cgagacgcga gtttgccggt ggtgcgaaca atagagcgac catgaccttg aaggtgagac
     7861 gcgcataacc gctagagtac tttgaagagg aaacagcaat agggttgcta ccagtataaa
     7921 tagacaggta catacaacac tggaaatggt tgtctgtttg agtacgcttt caattcattt
     7981 gggtgtgcac tttattatgt tacaatatgg aagggaactt tacacttctc ctatgcacat
     8041 atattaatta aagtccaatg ctagtagaga aggggggtaa cacccctccg cgctcttttc
     8101 cgattttttt ctaaaccgtg gaatatttcg gatatccttt tgttgtttcc gggtgtacaa
     8161 tatggacttc ctcttttctg gcaaccaaac ccatacatcg ggattcctat aataccttcg
     8221 ttggtctccc taacatgtag gtggcggagg ggagatatac aatagaacag ataccagaca
     8281 agacataatg ggctaaacaa gactacacca attacactgc ctcattgatg
//
